WO2019129144A1 - Modèle de classification pour détecter le degré de malignité d'un néoplasme oesophagien et/ou d'un néoplasme gastrique et son utilisation - Google Patents

Modèle de classification pour détecter le degré de malignité d'un néoplasme oesophagien et/ou d'un néoplasme gastrique et son utilisation Download PDF

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WO2019129144A1
WO2019129144A1 PCT/CN2018/124373 CN2018124373W WO2019129144A1 WO 2019129144 A1 WO2019129144 A1 WO 2019129144A1 CN 2018124373 W CN2018124373 W CN 2018124373W WO 2019129144 A1 WO2019129144 A1 WO 2019129144A1
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imprinted
imprinted gene
gene
genes
expression amount
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成彤
周宁
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Lisen Imprinting Diagnostics Inc
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Lisen Imprinting Diagnostics Inc
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer

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  • the present disclosure relates to the field of biotechnology, and relates to the field of genetic diagnosis, such as the field of genetic diagnosis, such as a hierarchical model and its application, such as a hierarchical model for detecting the degree of benign and malignant esophageal tumors and/or gastric tumors, and applications thereof, for example, A device and application of a hierarchical model of imprinted genes in detecting the degree of benign and malignant esophageal tumors and/or gastric tumors and their composition.
  • Gastric cancer and esophageal cancer are two common types of digestive tract cancers. In 2012, there were 952,000 new cases of gastric cancer, 723,000 deaths, 456,000 new cases of esophageal cancer, and 400,000 deaths. China is a high-risk area for gastric cancer and esophageal cancer. The annual incidence of new cases and deaths of gastric cancer and esophageal cancer account for about 50% of the world. The 5-year survival rate of early gastric cancer and esophageal cancer is over 90%, while the 5-year survival rate of advanced gastric cancer and esophageal cancer is less than 30%. Therefore, early diagnosis of gastric cancer and esophageal cancer is very important to save patients' lives. However, gastric cancer and esophageal cancer usually have no obvious symptoms in the early stage. The detection rate in China is less than 10%. More than half of the patients are already in the advanced stage when they find gastric cancer or esophageal cancer.
  • intraepithelial neoplasia is a precancerous lesion of the stomach, which can be classified into low-grade intraepithelial neoplasia and high-grade intraepithelial neoplasia depending on the degree of hyperplasia.
  • Low-grade intraepithelial neoplasia is about 20% likely to develop gastric cancer, while more than 70% of high-grade intraepithelial neoplasia may develop into gastric cancer.
  • the pathological distinction between low-grade and high-grade intraepithelial neoplasia is mainly subjective judgment by pathologists based on morphology.
  • CN 106609037 A discloses a detection kit for diagnosing gastric cancer patients based on a plurality of genes, wherein the kit includes a primer for diagnosing gastric cancer, which can be used for diagnosing gastric cancer, but the detection thereof is not accurate enough to distinguish which specific gastric cancer is specific. stage.
  • the pathological diagnosis of benign and malignant cells by traditional pathology is based on the relationship between cell size, morphology, invasiveness and surrounding cellular tissues. It has great limitations on the discovery of early changes in cells (cancer), so the method of cancer diagnosis at the cellular level has once become a research hotspot. With the continuous research in the field of molecular biology, more and more molecular detection techniques are applied to cancer diagnosis.
  • the production of cancer is an uncontrolled cell growth/splitting caused by epigenetic changes and genetic variations that accumulate over time.
  • Traditional pathological diagnosis is based on the size, morphology and structural variation of cells and tissues, thereby making a benign and malignant judgment of esophageal and gastric tumors.
  • molecular detection techniques With the development and deepening of molecular biology, more and more molecular detection techniques have been applied to the detection of esophageal cancer and gastric cancer. From the analysis of the development of cancer, molecular changes (epigenetics and genetics) are much earlier than changes in cell morphology and tissue structure. Therefore, molecular biology testing is more sensitive to early detection of cancer.
  • the current diagnosis of esophageal cancer and gastric cancer requires a new detection system and detection model.
  • the molecular markers of esophageal cancer and gastric cancer at the cellular level are analyzed to provide more accurate pre-diagnosis and Diagnostic information.
  • the present disclosure provides an imprinted gene grading model and its application.
  • the present disclosure provides an imprinted gene grading model for esophageal tumors and/or gastric tumors, the model calculating the amount of imprinted gene deletion expression, the imprinted gene copy number abnormal expression amount, and the total expression amount of the imprinted gene Changes in the esophageal tumor and/or gastric tumor classify the expression status of the imprinted gene;
  • the imprinted gene is any one or a combination of at least two of Z1, Z11 or Z16, the imprinting gene Z1 is Gnas, the imprinting gene Z11 is Grb10, and the imprinting gene Z16 is Snrpn/Snurf.
  • the imprinted (trace) deletion refers to the activation (demethylation) of the allele in the imprinted (spot) gene that was originally in a silent state, and is the most common and early epigenetic change in cancer, and This property can be used as a pathological marker.
  • the proportion of imprinted deletions is very low, and the imprinted gene and the imprinted gene are simultaneously a concept, indicating the same meaning, and can be replaced.
  • the public has found that the sensitivity of the diagnosis of esophageal cancer can reach 90.4% by calculating the amount of imprinted gene deletion and the abnormal expression of imprinted gene copy number in any esophageal tumor and/or gastric tumor of Z1, Z11 and Z16. Above, the sensitivity of diagnosis to gastric cancer can reach 67.6% or more.
  • any one of Z1, Z11 and Z16 can be detected if a preliminary detection detects only one imprinted gene.
  • the most preferred imprinting gene for detecting esophageal cancer is Z1.
  • the most preferred imprinting gene for detecting gastric cancer is Z11.
  • the inventors found that the detection of a Z1 imprinted gene alone can be 96.3% sensitive to esophageal cancer, and a Z11 imprinted gene can be detected alone, and the sensitivity for diagnosis of esophageal cancer can reach 92.5%. Detection of a Z16 imprinted gene, the sensitivity of diagnosis of esophageal cancer can reach 90.4%.
  • the inventors found that the detection of a Z1 imprinted gene alone can achieve a sensitivity of 80.6% for gastric cancer, and a Z11 imprinted gene can be detected alone, and the sensitivity for diagnosis of gastric cancer can reach 88.9%.
  • the Z16 imprinted gene has a sensitivity of 67.6% for the diagnosis of gastric cancer.
  • the combination may be any two of Z1, Z11 and Z16, for example the combination may be a combination of Z1 and Z11, Z1 and Z16 Combination or combination of Z11 and Z16.
  • the inventors have found that by calculating the amount of imprinted gene deletion and the abnormal expression level of imprinted gene copy number of two or more imprinted genes, the sensitivity can be further improved, and the combination of two imprinted genes of the imprinted gene can be detected to diagnose esophageal cancer.
  • Sensitivity can reach more than 98.1%.
  • the sensitivity of diagnosis to esophageal cancer can reach 98.1%.
  • the sensitivity of diagnosis to esophageal cancer can reach 99.2%.
  • the sensitivity of diagnosis to esophageal cancer can reach 99.2%;
  • the inventors have found that by calculating the amount of imprinted gene deletion and the abnormal expression of imprinted gene copy number of two or more imprinted genes, the sensitivity can be further improved, and the combination of two imprinted genes of the imprinted gene can be detected, which is sensitive to the diagnosis of gastric cancer.
  • the degree can reach more than 94.1%.
  • the sensitivity of diagnosis to gastric cancer can reach 97.1%.
  • the sensitivity of diagnosis to gastric cancer can reach 97.2%, and the detection of Z11 and Z16 is detected.
  • the diagnostic sensitivity to gastric cancer can reach 94.1%.
  • the imprinting gene further comprises any one or a combination of at least two of Z3, Z4, Z5, Z6, Z8, Z10 or Z13; wherein the imprinting gene Z3 is Peg10, the imprint The gene Z4 is Igf2r, the imprinted gene Z5 is Mest, the imprinted gene Z6 is Plagl1, the imprinted gene Z8 is Dcn, the imprinted gene Z10 is Gatm, and the imprinted gene Z13 is Sgce.
  • the method for calculating the imprinted gene is: calculating a combination of imprinted genes, the imprinted genes are ten of Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 A combination of genes.
  • the imprinted gene is deleted after the cells are subjected to hematoxylin staining, and there are two red/brown marks in the nucleus, and the abnormal copy number of the imprinted gene is that after the cells are subjected to hematoxylin staining, there are two or more red/brown marks in the nucleus.
  • the copy number abnormality is caused by abnormal gene replication of cancer cells, resulting in the expression of this gene as triploid or even higher polyploid.
  • the hematoxylin-stained label is selected from, but not limited to, red or brown, and staining with other colors can also be used for calculation of the total expression level of the imprinted gene, the amount of imprinted gene deletion, and the abnormal expression level of the imprinted gene copy number.
  • the formula for calculating the total expression amount of the imprinted gene, the expression amount of the imprinted gene, and the abnormal expression amount of the imprinted gene are as follows:
  • Imprinted gene deletion gene expression level (LOI) c / (b + c + d) ⁇ 100%;
  • a is a cell nucleus in which there is no label in the nucleus and no imprinted gene is expressed after the hematoxylin staining of the cell; and b is a red/brown mark in the nucleus after the hematoxylin staining of the cell, and the imprinting gene exists.
  • the nucleus; the c is a hematoxylin staining of the cells, there are two red/brown marks in the nucleus, and the nucleus of the imprinted gene is deleted; and the d is a hematoxylin staining of the cells, and there are more than two red/brown marks in the nucleus.
  • imprinted gene copy number abnormal cell nucleus is a cell nucleus in which there is no label in the nucleus and no imprinted gene is expressed after the hematoxylin staining of the cell
  • b is a red/brown mark in the nucleus after the hematoxylin staining of the cell,
  • the hematoxylin-stained label is selected from, but not limited to, red or brown, and staining with other colors can also be used for imprinting gene deletion expression, imprinted gene copy number abnormal expression, and imprinted gene total expression. The calculation of the amount.
  • the probe is amplified by in situ hybridization, and Hemotoxy (hematoxylin) nuclei are used to amplify the signal, and under the 40 ⁇ or 60 ⁇ microscope, the presence of the imprinted gene in each nucleus, the imprinted gene is deleted, or The copy number is abnormal, and the degree of benign and malignant tumors of the sample is determined by calculating the expression level of the imprinted gene deletion gene, the abnormal expression level of the imprinted gene copy number, and the total expression level of the imprinted gene; since the slice is only 10 ⁇ m, the nuclei seen under the microscope are approximately 20% are incomplete nuclei, which means that there is a possibility of partial false negatives.
  • Hemotoxy hematoxylin
  • the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the imprinted gene total expression amount are divided into five different levels.
  • At least 1200 cells are counted by each probe in the region where the sample is most positive, for ten of Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16
  • the imprinted gene's imprinted gene deletion expression amount, imprinted gene copy number abnormal expression amount, and imprinted gene total expression amount were divided into five different levels.
  • the five different levels of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of Z1 are:
  • the imprinted gene deletion expression amount of the imprinted gene Z1 is less than 16%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is less than 2.5%, or the total expression amount of the imprinted gene Z1 is less than 35% One or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z1 is 16-20%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is 2.5-4%, or the total expression amount of the imprinted gene Z1 is 35 Any one of -40% or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z1 is 20-25%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is 4-6.5%, or the total expression amount of the imprinted gene Z1 is 40. -50% of any one or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z1 is 25-30%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is 6.5-8%, or the total expression amount of the imprinted gene Z1 is 50 -65% of any one or a combination of at least two;
  • Grade IV the imprinted gene deletion expression amount of the imprinted gene Z1 is greater than 30%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is greater than 8%, or the total expression amount of the imprinted gene Z1 is greater than 65% One or a combination of at least two.
  • the five different levels of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount for Z3 and Z13 are:
  • the imprinted gene deletion expression amount of the imprinted genes Z3 and Z13 is less than 10%, the imprinted gene copy number abnormal expression amount of the imprinted genes Z3 and Z13 is less than 1% or the total expression amount of the imprinted genes Z3 and Z13 Any one of less than 25% or a combination of at least two;
  • the imprinted gene Z3 and Z13 have an imprinted gene deletion expression of 10-15%, the imprinted genes Z3 and Z13 have an imprinted gene copy number abnormal expression of 1-2% or the imprinted genes Z3 and Z13
  • the total expression amount is any one of 25-30% or a combination of at least two;
  • the imprinted gene Z3 and Z13 have an imprinted gene deletion expression of 15-25%, the imprinted genes Z3 and Z13 have an imprinted gene copy number abnormal expression of 2-3% or the imprinted genes Z3 and Z13
  • the total expression amount is any one of 30-45% or a combination of at least two;
  • the imprinted gene Z3 and Z13 have an imprinted gene deletion expression of 25-30%, the imprinted genes Z3 and Z13 have an imprinted gene copy number abnormal expression of 3-5% or the imprinted genes Z3 and Z13
  • the total expression amount is any one of 45-55% or a combination of at least two;
  • Grade IV the imprinted gene deletion expression amount of the imprinted genes Z3 and Z13 is greater than 30%, the imprinted gene copy number abnormal expression amount of the imprinted genes Z3 and Z13 is greater than 5% or the total expression amount of the imprinted genes Z3 and Z13 Any one of more than 55% or a combination of at least two;
  • the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of the imprinted genes Z3 and Z13 are independent of each other.
  • the five different levels of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount for Z4, Z5, Z6, Z10, and Z11 are:
  • the imprinted gene Z4, Z5, Z6, Z10 and Z11 have an imprinted gene deletion expression of less than 16%, and the imprinted genes Z4, Z5, Z6, Z10 and Z11 have an abnormal expression number of the imprinted gene of less than 1.5%. Or the total expression level of the imprinted genes Z4, Z5, Z6, Z10 and Z11 is less than 30% or a combination of at least two;
  • the imprinted gene Z4, Z5, Z6, Z10, and Z11 have an imprinted gene expression level of 16-20%, and the imprinted genes Z4, Z5, Z6, Z10, and Z11 have abnormal expression levels of the imprinted gene copy number. 1.5-2.5% or a total expression amount of the imprinting genes Z4, Z5, Z6, Z10 and Z11 of any one of 30-40% or a combination of at least two;
  • the imprinted gene Z4, Z5, Z6, Z10 and Z11 have an imprinted gene expression level of 20-25%, and the imprinted genes Z4, Z5, Z6, Z10 and Z11 have an abnormal expression level of the imprinted gene copy number.
  • 2.5-4% or a total expression amount of the imprinted genes Z4, Z5, Z6, Z10 and Z11 is any one of 40-50% or a combination of at least two;
  • the imprinted gene Z4, Z5, Z6, Z10 and Z11 have an imprinted gene expression level of 25-30%, and the imprinted genes Z4, Z5, Z6, Z10 and Z11 have an abnormal expression level of the imprinted gene copy number. 4-6% or a total expression amount of the imprinting genes Z4, Z5, Z6, Z10 and Z11 is any one of 50-65% or a combination of at least two;
  • the imprinted gene Z4, Z5, Z6, Z10 and Z11 have an imprinted gene expression level greater than 30%, and the imprinted genes Z4, Z5, Z6, Z10 and Z11 have an abnormal expression number of the imprinted gene greater than 6%. Or the total expression level of the imprinted genes Z4, Z5, Z6, Z10 and Z11 is greater than 65% or a combination of at least two;
  • the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of the imprinted genes Z4, Z5, Z6, Z10, and Z11 are independent of each other.
  • the five different grades of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of Z16 are:
  • any of the imprinted gene Z16 has an imprinted gene deletion expression of less than 10%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression amount of less than 1%, or the imprinted gene Z16 has a total expression amount of less than 25%.
  • the imprinted gene Z16 has an imprinted gene deletion expression of 10-15%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression of 1-2% or the imprinted gene Z16 has a total expression of 25%. Any one of -30% or a combination of at least two;
  • the imprinted gene Z16 has an imprinted gene deletion expression of 15-25%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression of 2-4% or the imprinted gene Z16 has a total expression of 30%. Any one of -40% or a combination of at least two;
  • the imprinted gene Z16 has an imprinted gene deletion expression amount of 25-30%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression amount of 4-6% or the imprinted gene Z16 has a total expression amount of 40%. Any one of -55% or a combination of at least two;
  • Grade IV the imprinting gene deletion expression amount of the imprinting gene Z16 is greater than 30%, the imprinting gene copy number abnormal expression amount of the imprinting gene Z16 is greater than 6%, or the total expression amount of the imprinting gene Z16 is greater than 55% One or a combination of at least two;
  • the five different grades of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of Z16 are:
  • the imprinted gene Z16 has an imprinted gene deletion expression amount of less than 15%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression amount of less than 1.5%, or the imprinted gene Z16 has a total expression amount of less than 30%.
  • the imprinted gene Z16 has an imprinted gene deletion expression of 15-20%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression of 1.5-4% or the imprinted gene Z16 has a total expression of 30%. Any one of -40% or a combination of at least two;
  • the imprinted gene Z16 has an imprinted gene deletion expression of 20-25%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression of 4-7% or the imprinted gene Z16 has a total expression of 40%. -50% of any one or a combination of at least two;
  • the imprinted gene Z16 has an imprinted gene deletion expression amount of 25-30%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression amount of 7-10% or the imprinted gene Z16 has a total expression amount of 50%. -60% of any one or a combination of at least two;
  • Grade IV any of the imprinted gene Z16 having an imprinted gene deletion expression amount greater than 30%, the imprinted gene Z16 imprinted gene copy number abnormal expression amount being greater than 10%, or the imprinted gene Z16 total expression amount being greater than 60% One or a combination of at least two.
  • the five different grades of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of Z8 are:
  • the imprinted gene Z8 has an imprinted gene deletion expression amount of less than 16%, the imprinted gene Z8 has an imprinted gene copy number abnormal expression amount of less than 2.5%, or the imprinted gene Z8 has a total expression amount of less than 35%.
  • the imprinted gene Z8 has an imprinted gene deletion expression amount of less than 16%, the imprinted gene Z8 has an imprinted gene copy number abnormal expression amount of less than 2.5%, or the imprinted gene Z8 has a total expression amount of less than 35%.
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is 16-20%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is 2.5-4%, or the total expression amount of the imprinted gene Z8 is 35 Any one of -40% or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is 20-25%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is 4-6.5%, or the total expression amount of the imprinted gene Z8 is 40 -50% of any one or a combination of at least two;
  • the imprinted gene Z8 has an imprinted gene deletion expression amount of 25-30%, the imprinted gene Z8 has an imprinted gene copy number abnormal expression amount of 6.5-8% or the imprinted gene Z8 has a total expression amount of 50%. -65% of any one or a combination of at least two;
  • Grade IV the imprinted gene deletion expression amount of the imprinted gene Z8 is greater than 30%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is greater than 8%, or the total expression amount of the imprinted gene Z8 is greater than 65% One or a combination of at least two;
  • the five different grades of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of Z8 are:
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is less than 10%, the abnormal expression level of the imprinted gene copy number of the imprinted gene Z8 is less than 1%, or the total expression amount of the imprinted gene Z8 is less than 25%.
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is less than 10%, the abnormal expression level of the imprinted gene copy number of the imprinted gene Z8 is less than 1%, or the total expression amount of the imprinted gene Z8 is less than 25%.
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is 10-15%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is 1-2%, or the total expression amount of the imprinted gene Z8 is 25 Any one of -30% or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is 15-25%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is 2-3%, or the total expression amount of the imprinted gene Z8 is 30 -45% of any one or a combination of at least two;
  • the imprinted gene Z8 has an imprinted gene deletion expression amount of 25-30%, the imprinted gene Z8 has an imprinted gene copy number abnormal expression amount of 3-5% or the imprinted gene Z8 has a total expression amount of 45%. Any one of -55% or a combination of at least two;
  • Grade IV the imprinted gene deletion expression amount of the imprinted gene Z8 is greater than 30%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is greater than 5%, or the total expression amount of the imprinted gene Z8 is greater than 55% One or a combination of at least two.
  • the present disclosure provides a device for detecting the degree of benign and malignant esophageal tumors and/or gastric tumors, which employs the above-described imprinted gene grading model, comprising the following units:
  • sampling unit obtaining a sample to be tested
  • Probe design unit design specific primers according to the imprinted gene sequence
  • the analysis unit calculates the total expression amount of the imprinted gene, the imprinted gene deletion expression amount, and the abnormal expression amount of the imprinted gene copy number, and the imprinted gene deletion expression amount and the imprinted gene copy number abnormality are obtained by the imprinted gene classification model.
  • the level of expression and the total expression level of the imprinted gene are used to determine the degree of benign and malignant esophageal tumors and/or gastric tumors.
  • the imprinted gene is deleted after the cells are subjected to hematoxylin staining, and there are two red/brown labeled nuclei in the nucleus.
  • the abnormal copy number of the imprinted gene is that after the cells are subjected to hematoxylin staining, there are more than two red/brown marks in the nucleus.
  • the copy number abnormality is caused by abnormal gene replication of cancer cells, resulting in the expression of this gene as triploid or even higher polyploid.
  • the hematoxylin-stained label is selected from, but not limited to, red or brown, and staining with other colors can also be used for calculation of total expression of imprinted gene, amount of imprinted gene deletion, and abnormal expression level of imprinted gene copy number.
  • the detection system is used for early and intuitive observation of changes in the imprinted and traced genes of the esophagus and gastric tumors at the cellular and tissue levels to determine the benign and malignant extent of the tumor, and to provide the most favorable treatment for patients with early esophageal and gastric tumors. opportunity.
  • the present disclosure provides a method for detecting the degree of benign and malignant esophageal tumors and/or gastric tumors, using the imprinted gene staging model, comprising the steps of:
  • the analysis unit by calculating the imprinted gene deletion expression amount and the imprinted gene copy number abnormal expression amount, by the imprinted gene grading model according to any one of claims 1 to 9, thereby imprinting the gene deletion expression amount and the imprinted gene
  • the level of abnormal expression of copy number is used to diagnose the degree of benign and malignant esophageal tumors and/or gastric tumors.
  • the sample to be tested described in step (1) is derived from human tissues and/or cells.
  • the sample to be tested is feasible as long as the RNA is subjected to a timely fixation process, and can be selected by a person skilled in the art as needed, and is not particularly limited herein.
  • the sample to be tested in the present disclosure includes paraffin sections of the tissue and esophageal mucosa falling off. Any one or combination of at least two of the cell or gastroscopic biopsy samples.
  • the specific operation procedure of the paraffin section of the tissue is to obtain a human tumor tissue sample, which is fixed in 10% neutral formalin in time, embedded in paraffin, cut into 10 ⁇ m thick, and made into a tissue film with a positively charged slide; Only 10 ⁇ m thick, so some of the microscopically seen nuclei are incomplete, so some false negative gene deletions will occur.
  • the specific operation procedure of the esophageal mucosal exfoliated cells is to obtain mucosal exfoliated cells by esophageal pull net examination, and timely fix with 10% neutral formalin.
  • the specific operation procedure of the gastroscope biopsy sample is to obtain human cells of the esophagus or stomach under the gastroscope, and the 10th neutral formalin can be fixed in time.
  • the sampling process is simple. Compared with the blood circulation characteristics, esophageal mucosal exfoliated cells and gastroscopic biopsy can be located. Esophageal mucosal exfoliated cells and gastroscopic biopsy have special effects as experimental samples. The advantages.
  • the sample to be tested is an esophageal mucosal exfoliated cell and/or a gastroscopic biopsy sample.
  • the imprinting genes are Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16, the imprinting gene Z1 is Gnas, and the imprinting gene Z3 is Peg10, The imprinted gene Z4 is Igf2r, the imprinted gene Z5 is Mest, the imprinted gene Z6 is Plagl1, the imprinted gene Z8 is Dcn, the imprinted gene Z10 is Gatm, the imprinted gene Z11 is Grb10, and the imprinted gene Z13 is Sgce, and the imprinting gene Z16 is Snrpn/Snurf.
  • the imprinted genes Z1 (Gnas), Z3 (Peg10), Z4 (Igf2r), Z5 (Mest), Z6 (Plagl1), Z8 (Dcn), Z10 (Gatm), Z11 (Grb10), Z13 (Sgce), Z16 (Snrpn/Snurf) has different degrees of expression in normal tumor cell tissues, and the expression and imprinting state will change significantly when malignant lesions occur.
  • the design probes are based on the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16, namely Gnas, Peg10, Igf2r, Mest, Plagl, Dcn, Gatm, Grb10, Sgce and Snrpn/ Designed by Snurf, a sequence was selected as a probe within the internal loop of each gene, and the specific probe was designed by Advanced Cell Diagnostics.
  • the in situ hybridization employs an RNAscope in situ hybridization method.
  • the RNAscope in situ hybridization method uses a single or multi-channel colorimetric kit or a single or multi-channel fluorescent kit, preferably a single channel red/brown color kit or multiple channels. Fluorescent kit.
  • the multi-channel coloring kit or the multi-channel fluorescent kit comprises two or more channels of coloring kits or fluorescent kits, and the two-channel coloring kit or multi-channel fluorescent kit can use two The combined expression of imprinted gene probes or imprinted genes and other genes is even the combined expression of multiple imprinted genes and non-imprinted genes.
  • the formula for calculating the total expression amount of the imprinted gene, the amount of the imprinted gene, and the abnormal expression amount of the imprinted gene in the model are as follows:
  • Imprinted gene deletion gene expression level (LOI) c / (b + c + d) ⁇ 100%;
  • a is a cell nucleus in which there is no label in the nucleus and no imprinted gene is expressed after the hematoxylin staining of the cell; and b is a red/brown mark in the nucleus after the hematoxylin staining of the cell, and the imprinting gene exists.
  • the nucleus; the c is a hematoxylin staining of the cells, there are two red/brown marks in the nucleus, and the nucleus of the imprinted gene is deleted; and the d is a hematoxylin staining of the cells, and there are more than two red/brown marks in the nucleus.
  • imprinted gene copy number abnormal cell nucleus is a cell nucleus in which there is no label in the nucleus and no imprinted gene is expressed after the hematoxylin staining of the cell
  • b is a red/brown mark in the nucleus after the hematoxylin staining of the cell,
  • the hematoxylin-stained label is selected from, but not limited to, red or brown, and staining with other colors can also be used for calculation of the total expression level of the imprinted gene, the amount of imprinted gene deletion, and the abnormal expression level of the imprinted gene copy number.
  • the probe was amplified by in situ hybridization, and Hemotoxy (hematoxylin) nuclei were used to amplify the signal, and under the 40 ⁇ or 60 ⁇ microscope, the presence of imprinted genes, imprinted gene deletion or copy number abnormality in each nucleus was determined, and the imprint was calculated.
  • the total gene expression level, the amount of the imprinted gene deletion gene, and the gene expression amount of the imprinted gene copy number abnormality were used to determine the degree of benign and malignant tumors of the sample. Since the section is only 10 microns, about 20% of the nuclei seen under the microscope are incomplete nuclei, which means that there is a possibility of partial false negatives.
  • the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount are divided into five different levels.
  • the five different grades count at least 1200 cells in the region where the probe is most positive for each probe, for ten of Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16
  • the imprinted gene deletion expression amount of the imprinted gene, the abnormal expression level of the imprinted gene copy number, and the total expression amount of the imprinted gene were separately divided.
  • the five different levels of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount for Z1 are:
  • the imprinted gene deletion expression amount of the imprinted gene Z1 is less than 16%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is less than 2.5%, or the total expression amount of the imprinted gene Z1 is less than 35% One or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z1 is 16-20%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is 2.5-4%, or the total expression amount of the imprinted gene Z1 is 35 Any one of -40% or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z1 is 20-25%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is 4-6.5%, or the total expression amount of the imprinted gene Z1 is 40. -50% of any one or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z1 is 25-30%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is 6.5-8%, or the total expression amount of the imprinted gene Z1 is 50 -65% of any one or a combination of at least two;
  • Grade IV the imprinted gene deletion expression amount of the imprinted gene Z1 is greater than 30%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z1 is greater than 8%, or the total expression amount of the imprinted gene Z1 is greater than 65% One or a combination of at least two.
  • the five different levels of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount for Z3 and Z13 are:
  • the imprinted gene deletion expression amount of the imprinted genes Z3 and Z13 is less than 10%, the imprinted gene copy number abnormal expression amount of the imprinted genes Z3 and Z13 is less than 1% or the total expression amount of the imprinted genes Z3 and Z13 Any one of less than 25% or a combination of at least two;
  • the imprinted gene Z3 and Z13 have an imprinted gene deletion expression of 10-15%, the imprinted genes Z3 and Z13 have an imprinted gene copy number abnormal expression of 1-2% or the imprinted genes Z3 and Z13
  • the total expression amount is any one of 25-30% or a combination of at least two;
  • the imprinted gene Z3 and Z13 have an imprinted gene deletion expression of 15-25%, the imprinted genes Z3 and Z13 have an imprinted gene copy number abnormal expression of 2-3% or the imprinted genes Z3 and Z13
  • the total expression amount is any one of 30-45% or a combination of at least two;
  • the imprinted gene Z3 and Z13 have an imprinted gene deletion expression of 25-30%, the imprinted genes Z3 and Z13 have an imprinted gene copy number abnormal expression of 3-5% or the imprinted genes Z3 and Z13
  • the total expression amount is any one of 45-55% or a combination of at least two;
  • Grade IV the imprinted gene deletion expression amount of the imprinted genes Z3 and Z13 is greater than 30%, the imprinted gene copy number abnormal expression amount of the imprinted genes Z3 and Z13 is greater than 5% or the total expression amount of the imprinted genes Z3 and Z13 Any one of more than 55% or a combination of at least two;
  • the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of the imprinted genes Z3 and Z13 are independent of each other.
  • the five different levels of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount for Z4, Z5, Z6, Z10, and Z11 are:
  • the imprinted gene Z4, Z5, Z6, Z10 and Z11 have an imprinted gene deletion expression of less than 16%, and the imprinted genes Z4, Z5, Z6, Z10 and Z11 have an abnormal expression number of the imprinted gene of less than 1.5%. Or the total expression level of the imprinted genes Z4, Z5, Z6, Z10 and Z11 is less than 30% or a combination of at least two;
  • the imprinted gene Z4, Z5, Z6, Z10, and Z11 have an imprinted gene expression level of 16-20%, and the imprinted genes Z4, Z5, Z6, Z10, and Z11 have abnormal expression levels of the imprinted gene copy number. 1.5-2.5% or a total expression amount of the imprinting genes Z4, Z5, Z6, Z10 and Z11 of any one of 30-40% or a combination of at least two;
  • the imprinted gene Z4, Z5, Z6, Z10 and Z11 have an imprinted gene expression level of 20-25%, and the imprinted genes Z4, Z5, Z6, Z10 and Z11 have an abnormal expression level of the imprinted gene copy number.
  • 2.5-4% or a total expression amount of the imprinted genes Z4, Z5, Z6, Z10 and Z11 is any one of 40-50% or a combination of at least two;
  • the imprinted gene Z4, Z5, Z6, Z10 and Z11 have an imprinted gene expression level of 25-30%, and the imprinted genes Z4, Z5, Z6, Z10 and Z11 have an abnormal expression level of the imprinted gene copy number. 4-6% or a total expression amount of the imprinting genes Z4, Z5, Z6, Z10 and Z11 is any one of 50-65% or a combination of at least two;
  • the imprinted gene Z4, Z5, Z6, Z10 and Z11 have an imprinted gene expression level greater than 30%, and the imprinted genes Z4, Z5, Z6, Z10 and Z11 have an abnormal expression number of the imprinted gene greater than 6%. Or the total expression level of the imprinted genes Z4, Z5, Z6, Z10 and Z11 is greater than 65% or a combination of at least two;
  • the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of the imprinted genes Z4, Z5, Z6, Z10, and Z11 are independent of each other.
  • the five different grades of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of Z16 are:
  • any of the imprinted gene Z16 has an imprinted gene deletion expression of less than 10%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression amount of less than 1%, or the imprinted gene Z16 has a total expression amount of less than 25%.
  • the imprinted gene Z16 has an imprinted gene deletion expression of 10-15%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression of 1-2% or the imprinted gene Z16 has a total expression of 25%. Any one of -30% or a combination of at least two;
  • the imprinted gene Z16 has an imprinted gene deletion expression of 15-25%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression of 2-4% or the imprinted gene Z16 has a total expression of 30%. Any one of -40% or a combination of at least two;
  • the imprinted gene Z16 has an imprinted gene deletion expression amount of 25-30%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression amount of 4-6% or the imprinted gene Z16 has a total expression amount of 40%. Any one of -55% or a combination of at least two;
  • Grade IV the imprinting gene deletion expression amount of the imprinting gene Z16 is greater than 30%, the imprinting gene copy number abnormal expression amount of the imprinting gene Z16 is greater than 6%, or the total expression amount of the imprinting gene Z16 is greater than 55% One or a combination of at least two.
  • the five different grades of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of Z16 are:
  • the imprinted gene Z16 has an imprinted gene deletion expression amount of less than 15%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression amount of less than 1.5%, or the imprinted gene Z16 has a total expression amount of less than 30%.
  • the imprinted gene Z16 has an imprinted gene deletion expression of 15-20%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression of 1.5-4% or the imprinted gene Z16 has a total expression of 30%. Any one of -40% or a combination of at least two;
  • the imprinted gene Z16 has an imprinted gene deletion expression of 20-25%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression of 4-7% or the imprinted gene Z16 has a total expression of 40%. -50% of any one or a combination of at least two;
  • the imprinted gene Z16 has an imprinted gene deletion expression amount of 25-30%, the imprinted gene Z16 has an imprinted gene copy number abnormal expression amount of 7-10% or the imprinted gene Z16 has a total expression amount of 50%. -60% of any one or a combination of at least two;
  • Grade IV any of the imprinted gene Z16 having an imprinted gene deletion expression amount greater than 30%, the imprinted gene Z16 imprinted gene copy number abnormal expression amount being greater than 10%, or the imprinted gene Z16 total expression amount being greater than 60% One or a combination of at least two.
  • the five different grades of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of Z8 are:
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is less than 16%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is less than 2.5%, or the total expression amount of the imprinted gene Z8 is less than 35% One or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is 16-20%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is 2.5-4%, or the total expression amount of the imprinted gene Z8 is 35 Any one of -40% or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is 20-25%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is 4-6.5%, or the total expression amount of the imprinted gene Z8 is 40 -50% of any one or a combination of at least two;
  • the imprinted gene Z8 has an imprinted gene deletion expression amount of 25-30%, the imprinted gene Z8 has an imprinted gene copy number abnormal expression amount of 6.5-8% or the imprinted gene Z8 has a total expression amount of 50%. -65% of any one or a combination of at least two;
  • Grade IV the imprinted gene deletion expression amount of the imprinted gene Z8 is greater than 30%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is greater than 8%, or the total expression amount of the imprinted gene Z8 is greater than 65% One or a combination of at least two;
  • the five different grades of the imprinted gene deletion expression amount, the imprinted gene copy number abnormal expression amount, and the total expression amount of Z8 are:
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is less than 10%, the abnormal expression level of the imprinted gene copy number of the imprinted gene Z8 is less than 1%, or the total expression amount of the imprinted gene Z8 is less than 25%.
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is less than 10%, the abnormal expression level of the imprinted gene copy number of the imprinted gene Z8 is less than 1%, or the total expression amount of the imprinted gene Z8 is less than 25%.
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is 10-15%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is 1-2%, or the total expression amount of the imprinted gene Z8 is 25 Any one of -30% or a combination of at least two;
  • the imprinted gene deletion expression amount of the imprinted gene Z8 is 15-25%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is 2-3%, or the total expression amount of the imprinted gene Z8 is 30 -45% of any one or a combination of at least two;
  • the imprinted gene Z8 has an imprinted gene deletion expression amount of 25-30%, the imprinted gene Z8 has an imprinted gene copy number abnormal expression amount of 3-5% or the imprinted gene Z8 has a total expression amount of 45%. Any one of -55% or a combination of at least two;
  • Grade IV the imprinted gene deletion expression amount of the imprinted gene Z8 is greater than 30%, the imprinted gene copy number abnormal expression amount of the imprinted gene Z8 is greater than 5%, or the total expression amount of the imprinted gene Z8 is greater than 55% One or a combination of at least two.
  • the imprinted gene Z16 was uniformly expressed in the stroma and epithelial cells of gastric tumor tissues, but was significantly different in the early stage of gastric cancer before the clinical stage II stage and the advanced stage gastric cancer of the clinical stage III-IV stage.
  • the degree of benign and malignant diagnosis of esophageal tumors is classified into benign tumors, esophageal cancer potential, early esophageal cancer, meta-esophageal cancer, and advanced esophageal cancer.
  • the result of the diagnosis of the degree of benign and malignant esophagus is the imprinted gene deletion expression amount and the imprinted gene copy number of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16.
  • the abnormal expression level was less than grade I, and the imprinted gene deletion level of no more than one imprinted gene in the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 was grade I and the imprinted gene Z1
  • the result of the diagnosis of the degree of benign and malignant esophagus is that the imprinted gene deletion expression level of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is grade I, imprinted
  • the abnormal expression level of the imprinted gene copy number of at least two imprinted genes of genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is grade I or imprinted genes Z1, Z3, Z4, Z5, Z6,
  • the imprinted gene deletion expression of no more than one imprinted gene in Z8, Z10, Z11, Z13 and Z16 is class II and the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16
  • the abnormal expression level of the imprinted gene copy number of more than one imprinted gene is any one of the second grade, and is the esophageal cancer potential;
  • the result of the diagnosis of the degree of benign and malignant esophagus is that the imprinted gene deletion expression level of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is Grade II, imprinted
  • the abnormal expression level of the imprinted gene copy number of at least two imprinted genes of genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is Grade II or imprinted genes Z1, Z3, Z4, Z5, Z6,
  • the imprinted gene deletion expression level of no more than one imprinted gene in Z8, Z10, Z11, Z13 and Z16 is grade III and the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 do not If the abnormal expression level of the imprinted gene copy number of more than one imprinted gene is in any of the grade III, it is early esophageal cancer;
  • the result of the diagnosis of the degree of benign and malignant esophagus is that the imprinted gene deletion expression level of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is grade III, imprinted
  • the abnormal expression level of imprinted gene copy number of at least two imprinted genes of genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is grade III or imprinted genes Z1, Z3, Z4, Z5, Z6,
  • the imprinted gene deletion expression of no more than one imprinted gene in Z8, Z10, Z11, Z13 and Z16 is IV and the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 do not If the abnormal expression level of the imprinted gene copy number of more than one imprinted gene is in any of the IV grades, it is a metaphase esophageal cancer;
  • the result of the diagnosis of the degree of benign and malignant esophagus is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 has a deletion level of IV or imprinted gene.
  • the abnormal expression level of the imprinted gene copy number of at least two of the Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13, Z8, Z10, Z11, Z13 and Z16 is grade IV, and is advanced esophageal cancer.
  • the diagnosis of benign and malignant gastric tumors is classified into benign gastric tumors, gastric cancer potential, early gastric cancer, metaphase gastric cancer, and advanced gastric cancer.
  • the results of the diagnosis of the degree of benign and malignant tumors of the gastric tumors are that the imprinted gene Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 have an imprinted gene expression level and an abnormal expression amount of the imprinted gene copy number are less than I.
  • the imprinted gene of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13, and Z16 has an imprinted gene deletion level of I and the imprinted genes Z1, Z3, Z4, Z5
  • the abnormal expression level of the imprinted gene copy number of not more than one imprinted gene in Z6, Z8, Z10, Z11, Z13, and Z16 is any one of grade I, and is a benign tumor;
  • the result of the diagnosis of the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of I, and the imprinted gene
  • the abnormal expression level of imprinted gene copy number of at least two imprinted genes of Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 is grade I
  • imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10 is expressed in the class I and has no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13.
  • the abnormal expression level of the imprinted gene copy number is grade I and the imprinted gene deletion expression amount of the imprinted gene Z16 or the abnormal expression level of the imprinted gene copy number is grade I or above, or the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10
  • the imprinting gene with no more than one imprinted gene in Z11 and Z13 has a deletion level of II and imprinted with no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13.
  • the abnormal expression level of the gene copy number is in any of the cases of grade II, which is the potential of gastric cancer;
  • the result of the diagnosis of the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of II, imprinted gene
  • the abnormal expression level of the imprinted gene copy number of at least two imprinted genes in Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 is Grade II or imprinted genes Z1, Z3, Z4, Z5, Z6, Z8,
  • the imprinted gene deletion expression level of no more than one imprinted gene in Z10, Z11 and Z13 is grade III and no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 If the abnormal expression level of the imprinted gene copy is in any of the grade III, it is early gastric cancer;
  • the result of the diagnosis of the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of III, imprinted gene
  • the abnormal expression level of imprinted gene copy number of at least 2 imprinted genes of Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 is grade III or imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10
  • the imprinting gene with no more than one imprinted gene in Z11 and Z13 is expressed at the IV level and the imprint of no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 If the abnormal expression level of the gene copy number is in any of the IV grades, it is a metaphase gastric cancer;
  • the result of diagnosing the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of IV or imprinted gene Z1.
  • the abnormal expression level of the imprinted gene copy number of at least two of the Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z6, Z8, Z10, Z11 and Z13 is grade IV, and is advanced gastric cancer.
  • the present disclosure provides the imprinted gene grading model and/or the device for esophageal tumor and gastric tumor detection.
  • the present disclosure provides the use of the imprinted gene grading model and/or the device for the preparation of a medicament or device for treating an esophageal tumor and/or a gastric tumor.
  • the degree of benign and malignant diagnosis of esophageal tumors is classified into benign tumors, esophageal cancer potential, early esophageal cancer, meta-esophageal cancer, and advanced esophageal cancer.
  • the results of the diagnosis of the degree of benign and malignant tumors of the esophageal tumor are that the imprinted gene Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 have an imprinted gene expression level and an abnormal expression amount of the imprinted gene copy number are less than I.
  • the imprinted gene of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13, and Z16 has an imprinted gene deletion level of I and the imprinted genes Z1, Z3, Z4, Z5
  • the abnormal expression level of the imprinted gene copy number of not more than one imprinted gene in Z6, Z8, Z10, Z11, Z13, and Z16 is any one of grade I, and is a benign tumor;
  • the result of the diagnosis of the degree of benign and malignant of the esophageal tumor is that the imprinted gene expression level of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is Grade I,
  • the abnormal expression level of the imprinted gene copy number of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is Grade I or imprinted genes Z1, Z3, Z4, Z5, Z6 In the Z8, Z10, Z11, Z13 and Z16, the imprinted gene deletion expression level of the imprinted gene is Grade II and the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16
  • the abnormal expression level of the imprinted gene copy number of no more than one imprinted gene is any one of the second grade, and is the esophageal cancer potential;
  • the result of the diagnosis of the degree of benign and malignant of the esophageal tumor is that the imprinted gene deletion expression level of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is II,
  • the abnormal expression level of the imprinted gene copy number of at least two imprinted genes of the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is Grade II or imprinted genes Z1, Z3, Z4, Z5, Z6 In the Z8, Z10, Z11, Z13 and Z16, the imprinted gene deletion expression level of the imprinted gene is III and the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 If the abnormal expression level of the imprinted gene copy number of no more than one imprinted gene is in any of the grade III, it is early esophageal cancer;
  • the result of the diagnosis of the degree of benign and malignant of the esophageal tumor is that the imprinted gene deletion expression level of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is III,
  • the abnormal expression level of imprinted gene copy number of at least two imprinted genes of imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is grade III or imprinted genes Z1, Z3, Z4, Z5, Z6 , the imprinting gene of Z8, Z10, Z11, Z13 and Z16 is not more than one imprinted gene, and the imprinted gene is expressed in the IV level and in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16. If the abnormal expression level of the imprinted gene copy number of no more than one imprinted gene is any of the IV grades, it is a metaphase esophageal cancer;
  • the result of the diagnosis of the degree of benign and malignant tumors of the esophageal tumor is that the imprinted gene of at least two of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 has an imprinted gene expression level of IV or imprinted.
  • the abnormal expression level of the imprinted gene copy number of at least two of the imprinted genes in the genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13, and Z16 is Grade IV, and is an advanced esophageal cancer.
  • the diagnosis of gastric tumors is classified into benign gastric tumors, gastric cancer potential, early gastric cancer, metaphase gastric cancer, and advanced gastric cancer.
  • the results of the diagnosis of the degree of benign and malignant tumors of the gastric tumors are that the imprinted gene Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 have an imprinted gene expression level and an abnormal expression amount of the imprinted gene copy number are less than I.
  • the imprinted gene of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13, and Z16 has an imprinted gene deletion level of I and the imprinted genes Z1, Z3, Z4, Z5
  • the abnormal expression level of the imprinted gene copy number of not more than one imprinted gene in Z6, Z8, Z10, Z11, Z13, and Z16 is any one of grade I, and is a benign tumor;
  • the result of the diagnosis of the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of I, and the imprinted gene
  • the abnormal expression level of imprinted gene copy number of at least two imprinted genes of Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 is grade I
  • imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10 is expressed in the class I and has no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13.
  • the abnormal expression level of the imprinted gene copy number is grade I and the imprinted gene deletion expression amount of the imprinted gene Z16 or the abnormal expression level of the imprinted gene copy number is grade I or above, or the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10
  • the imprinting gene with no more than one imprinted gene in Z11 and Z13 has a deletion level of II and imprinted with no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13.
  • the abnormal expression level of the gene copy number is in any of the cases of grade II, which is the potential of gastric cancer;
  • the result of the diagnosis of the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of II, imprinted gene
  • the abnormal expression level of the imprinted gene copy number of at least two imprinted genes in Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 is Grade II or imprinted genes Z1, Z3, Z4, Z5, Z6, Z8,
  • the imprinted gene deletion expression level of no more than one imprinted gene in Z10, Z11 and Z13 is grade III and no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 If the abnormal expression level of the imprinted gene copy is in any of the grade III, it is early gastric cancer;
  • the result of the diagnosis of the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of III, imprinted gene
  • the abnormal expression level of imprinted gene copy number of at least 2 imprinted genes of Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 is grade III or imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10
  • the imprinting gene with no more than one imprinted gene in Z11 and Z13 is expressed at the IV level and the imprint of no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 If the abnormal expression level of the gene copy number is in any of the IV grades, it is a metaphase gastric cancer;
  • the result of diagnosing the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of IV or imprinted gene Z1.
  • the abnormal expression level of the imprinted gene copy number of at least two of the Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z6, Z8, Z10, Z11 and Z13 is grade IV, and is advanced gastric cancer.
  • the present disclosure provides a marker for the diagnosis of advanced gastric cancer, the marker being the imprinting gene Z1 and the imprinting gene Z11;
  • imprinting gene Z1 is Gnas
  • imprinting gene Z11 is Grb10
  • the advanced gastric cancer is gastric cancer having a TNM rating of gastric cancer greater than T2N0M0.
  • the imprinted gene Z16 was uniformly expressed in the stroma and epithelial cells of gastric tumor tissues, but was significantly different in the early stage of gastric cancer before the clinical stage II stage and the advanced stage gastric cancer of the clinical stage III-IV stage. In early gastric cancer, especially in the period of intraepithelial neoplasia, 75% of the imprinted gene Z16 has an imprinted deletion, copy number abnormality and total expression level of more than grade I. With the deterioration of cancer, 90% of advanced gastric cancer samples The imprinted gene Z16 has an imprinted deletion, an abnormal copy number, and a total expression level of 0.
  • the imprinted gene Z16 combined with the imprinting genes Z1 and Z11 is a good diagnostic marker for the diagnosis of early gastric cancer, especially the transition from low-grade intraepithelial neoplasia to gastric cancer, but the imprinting gene Z16 is useful in advanced gastric cancer. Not big, only need Z1 and Z11 to make a diagnosis.
  • the detection model and the device are used to visually express the performance of the imprinted gene on the sample of the esophageal tumor and/or the gastric tumor patient, and the method for in situ labeling the imprinted gene Objective, intuitive, early, accurate detection of changes in the imprinted (trace) gene, and can provide quantitative models that make a significant contribution to the diagnosis of molecular pathology.
  • 1 is a pathological section of gastric cancer of the present disclosure by hematoxylin-stained nuclei, wherein the a is a hematoxylin-stained cell, and there is no label in the nucleus, and the imprinted gene is not expressed; the b is a cell nucleus after hematoxylin staining There is a red/brown mark in the presence of the imprinted gene; the c is the hematoxylin staining of the cells, there are two red/brown marks in the nucleus, and the imprinted gene is deleted; the d is the cell nucleus memory after hematoxylin staining In more than two red/brown markers, the imprinted gene copy number is abnormal;
  • Fig. 2(a) shows the expression status of 10 genes in the pathological section of grade 0 gastric tumor
  • Fig. 2(b) shows the expression status of 10 genes in the pathological section of grade I gastric cancer
  • Fig. 2(c) shows grade II gastric cancer.
  • the expression status of 10 genes in the pathological section Fig. 2(d) shows the expression status of 10 genes in the pathological section of grade III gastric cancer
  • Fig. 2(e) shows the expression status of 10 genes in the pathological section of grade IV gastric cancer. ;
  • Fig. 3(a) shows the intensity of imprinting deletion of gastric cancer genes Z1, Z11 and Z16
  • Fig. 3(b) shows the intensity of copy number abnormalities of gastric cancer Z1, Z11 and Z16 for gastric cancer
  • Fig. 3(c) is an imprint.
  • Fig. 3(d) are the intensities of the imprinted genes Z3, Z4, Z5, Z6, Z8, Z10 and Z13 on the gastric cancer
  • Fig. 3(e) is The intensity of the imprinting genes Z3, Z4, Z5, Z6, Z8, Z10 and Z13 on the copy number abnormality of gastric cancer
  • Fig. 3(e) is The intensity of the imprinting genes Z3, Z4, Z5, Z6, Z8, Z10 and Z13 on the copy number abnormality of gastric cancer
  • 3(f) is the total expression of the imprinted genes Z3, Z4, Z5, Z6, Z8, Z10 and Z13 on gastric cancer.
  • the intensity of the quantity wherein LOI is the expression level of the imprinted gene deletion gene, CNV is the gene expression quantity of the imprinted gene copy number abnormality, and TE is the total expression level of the imprinted gene;
  • Fig. 4(a) shows the intensity of the imprinting gene Z1 imprinting deletion, copy number abnormality and total expression amount
  • Fig. 4(b) shows the intensity of the imprinting gene Z11 imprinting deletion, copy number abnormality and total expression amount
  • FIG. 4(c) is The imprinting gene Z16 imprinted deletion, copy number abnormality and total expression level intensity
  • Figure 4 (d) is the imprinting gene Z3 imprint deletion, copy number abnormality and total expression level intensity
  • Figure 4 (e) is the imprinted gene Z4 imprint missing
  • Figure 4 (f) is the imprinting gene Z5 imprint deletion, copy number abnormality and total expression level intensity
  • Figure 4 (g) is imprinted gene Z6 imprint deletion, copy number abnormality and total expression
  • Figure 4 (h) is the intensity of the imprinting gene Z8 imprint deletion, copy number abnormality and total expression
  • Figure 4 (i) is the imprinting gene Z10 imprint deletion, copy number abnormality and total expression intensity
  • Figure 4 (j) The intensity of imprinting gene Z13 imprinting deletion, copy number abnormality and total expression amount, wherein LOI is the expression level of imprinted gene deletion gene, CNV is the gene expression amount of imprinted
  • Fig. 5(a) shows the distribution range and grading standard of imprinted gene Z1 in 37 cases of gastric tumor pathological sections, imprinting deletion and copy number abnormality
  • Fig. 5(b) is the imprinting gene Z11 applied to 37 cases of gastric tumor pathological section.
  • Fig. 5(c) shows the distribution range and grading standard of imprinting deletion and copy number abnormality in the pathological section of 37 gastric tumors with imprinting gene Z16
  • Fig. 5(d) The imprinted gene Z3 was applied to the pathological sections of 37 cases of gastric tumors, the distribution range and grading standard of imprinting deletion and copy number abnormality
  • Fig. 5(e) was the imprinted gene Z4 applied to 37 cases of gastric tumor pathological sections, imprinted deletion and The distribution range and grading standard of copy number abnormality
  • Fig. 5(f) shows the distribution range and grading standard of imprinting deletion and copy number abnormality in 37 pathological sections of gastric tumors
  • Fig. 5(g) is imprinted gene.
  • Z6 was applied to the pathological sections of 37 cases of gastric tumors, the distribution range and grading standard of imprinting deletion and copy number abnormality.
  • Figure 5(h) shows that imprinted gene Z8 was applied to 37 cases of gastric tumor pathological sections with imprinting deletion and copy number abnormality.
  • Figure 5 (i) is the distribution range and grading standard of imprinting gene Z10 applied to 37 gastric tumor pathological sections, imprinting deletion and copy number abnormality
  • Fig. 5 (j) is applied to imprinting gene Z13 in 37 cases In the pathological section of gastric tumors, the distribution range and grading criteria of imprinting deletion and copy number abnormality, wherein LOI is the expression level of imprinted gene deletion gene, CNV is the gene expression quantity of imprinted gene copy number abnormality, and TE is the total expression level of imprinted gene;
  • Fig. 6(a) shows the expression status of 10 genes in the pathological section of grade 0 esophageal tumor
  • Fig. 6(b) shows the expression status of 10 genes in the pathological section of grade I esophageal cancer
  • Fig. 6(c) is level II.
  • Fig. 6(d) shows the expression status of 10 genes in the pathological section of grade III esophageal cancer
  • Fig. 6(e) shows 10 cases in the pathological section of grade IV esophageal cancer.
  • Fig. 7(a) shows the intensity of imprinting defects in esophageal cancer by imprinting genes Z1, Z11 and Z16
  • Fig. 7(b) shows the intensity of copying abnormalities of esophageal cancer by imprinting genes Z1, Z11 and Z16
  • Fig. 7(c) For the intensity of the total expression of the imprinted genes Z1, Z11 and Z16 for esophageal cancer
  • Fig. 7(d) shows the intensity of imprinting defects of the imprinted genes Z3, Z4, Z5, Z6, Z8, Z10 and Z13 for esophageal cancer
  • Fig. 7(d) shows the intensity of imprinting defects of the imprinted genes Z3, Z4, Z5, Z6, Z8, Z10 and Z13 for esophageal cancer
  • Figure 8 (a) shows the intensity of the imprinted gene Z1 imprint deletion, copy number abnormality and total expression
  • Figure 8 (b) shows the imprinted gene Z11 imprint deletion, copy number abnormality and total expression intensity
  • Figure 8 (c) For the imprinting gene Z16 imprint deletion, copy number abnormality and total expression intensity
  • Figure 8 (d) is the imprinting gene Z3 imprint deletion, copy number abnormality and total expression intensity
  • Figure 8 (e) is the imprinted gene Z4 imprint deletion , copy number abnormality and total expression intensity
  • Figure 8 (f) is the imprinting gene Z5 imprint deletion, copy number abnormality and total expression intensity
  • Figure 8 (g) is the imprinted gene Z6 imprint deletion, copy number abnormality and total The intensity of the expression amount
  • 8(h) shows the intensity of the imprinted gene Z8 imprint deletion, copy number abnormality and total expression amount
  • Fig. 8(i) shows the intensity of the imprinted gene Z10 imprint deletion, copy number abnormality and total expression amount
  • 8(j) is the intensity of imprinting gene Z13 imprinting deletion, copy number abnormality and total expression amount, wherein LOI is the expression level of imprinted gene deletion gene, CNV is the gene expression amount of imprinted gene copy number abnormality, and TE is imprinted gene total expression. the amount;
  • Fig. 9(a) shows the distribution range and grading standard of imprinting deletion and copy number abnormality in 57 cases of esophageal tumor pathological sections
  • Fig. 9(b) is the imprinting gene Z11 applied to 57 esophage tumor pathological sections.
  • Fig. 9(c) shows the distribution range and grading standard of imprinting deletion and copy number abnormality in 57 pathological sections of esophageal tumor, which is imprinted gene Z16
  • Fig. 9(b) shows the distribution range and grading standard of imprinting deletion and copy number abnormality in 57 cases of esophageal tumor pathological sections
  • Fig. 9(b) is the imprinting gene Z11 applied to 57 esophage tumor pathological sections.
  • Fig. 9(c) shows the distribution range and grading standard of imprinting deletion and copy number abnormality in 57 pathological sections of esophageal tumor, which is imprinted gene Z16
  • the imprinting gene Z3 was applied to 57 cases of esophageal tumor pathological sections, the distribution range and grading standard of imprinting deletion and copy number abnormality
  • Fig. 9(e) was the imprinting gene Z4 applied to 57 esophage tumor pathological sections, imprinting deletion and The distribution range and grading standard of copy number abnormality
  • Fig. 9(f) shows the distribution range and grading standard of imprinting deletion and copy number abnormality in 57 pathological sections of esophageal tumor
  • the imprinting gene is shown in Fig.
  • FIG. 9(g) Z6 was applied to the pathological sections of esophageal tumors in 57 cases, and the distribution range and grading standard of imprinting deletion and copy number abnormality.
  • Figure 9(h) shows that imprinted gene Z8 was applied to 57 cases of esophageal tumor pathological sections, and the imprint was missing.
  • the distribution range and grading standard of copy number abnormality Fig. 9(i) shows the distribution range and grading standard of imprinting deletion and copy number abnormality in 57 pathological sections of esophageal tumor
  • Fig. 9(j) is the imprint Gene Z13 was applied to the pathological sections of esophageal tumors, and the distribution range and grading criteria of imprinting deletion and copy number abnormality.
  • LOI was the expression level of imprinted gene deletion gene
  • CNV was the gene expression level of imprinted gene copy number abnormality
  • TE was The total expression level of the imprinted gene.
  • Genomic imprinting is a way of gene regulation in epigenetics. It is characterized by methylation of alleles from a particular parent, such that one gene has only one allele and the other is in a state of gene silencing. This type of gene is called a blot (remember) gene. Deletion of the blot is an epigenetic change in which the allelic gene of the imprinted gene results in a silenced allele being activated and beginning to express the gene. Numerous studies have shown that this phenomenon (missing of blots) is ubiquitous in various types of cancer and occurs earlier than cell and tissue morphological changes. At the same time, in healthy cells, the proportion of imprinted deletions is extremely low, in sharp contrast to cancer cells. Therefore, the methylation status of the imprinted gene can be used as a pathological marker to analyze the abnormal state of the cell by a specific molecular detection technique.
  • the detection model and system of the present disclosure express the impression of the imprinted defect on the sample of the esophageal and gastric tumor patients in an intuitive manner, and objectively, intuitively, earlyly and accurately detect the imprint by the method of in situ labeling of the imprinted gene. (trace) changes in genes and can provide quantitative models that make a significant contribution to the diagnosis of esophageal and gastric tumors;
  • the detection device of the present invention can determine the degree of benign and malignant esophageal and gastric tumors and the extent of invasion of cancer cells by gastroscopic biopsy samples before operation in patients with esophagus and stomach tumors, thereby providing a basis for surgery and precise treatment, which is a diagnosis in the field of cell molecules.
  • the present disclosure can accurately determine the grades of esophageal and gastric tumors, and clearly classify the early lesions of esophageal and gastric tumors, ie, the degree of malignancy of intraepithelial neoplasia, by combination of imprinted genes, especially for early and advanced gastric cancer.
  • Accurate differentiation has greatly improved the early and clear diagnosis of esophageal cancer and gastric cancer, especially in early census and postoperative follow-up of cancer, especially for follow-up of suspected relapsed patients, which can win time and save lives for patients.
  • Major contribution
  • the disclosed detection method is different from the immunohistochemical method, and reduces false positives and other negative effects, and not only the targeted drugs which are silenced, eliminated, and rearranged by the discovery of the missing regions of the esophageal and gastric tumor-associated imprinted genes. Or technical methods that can be used to guide later treatment and medication.
  • the method for detecting the imprinted gene comprises the following steps:
  • tissue cell sections (10 ⁇ m) of gastric tumors and fixing them in a 10% neutral formalin solution to prevent RNA degradation for 24 hours, paraffin embedding (FFPE), The slide needs to be loaded with a positive charge, and the slice is baked in an oven at 40 ° C for more than 3 hours;
  • FFPE paraffin embedding
  • Design probe design specific primers according to the imprinted gene sequence
  • the design probe is based on the imprinting genes Z1 (Gnas), Z3 (Peg10), Z4 (Igf2r), Z5 (Mest), Z6 (Plagl1), Z8 (Dcn), Z10 (Gatm), Z11 (Grb10), Z13. (Sgce) and Z16 (Snrpn/Snurf) were designed to select a sequence within the internal loop of each gene as a probe. The specific probe was designed by Advanced Cell Diagnostics.
  • the formula for calculating the total expression of the imprinted gene, the amount of the imprinted gene, and the abnormal expression of the imprinted gene in the model are as follows:
  • Imprinted gene deletion gene expression level (LOI) c / (b + c + d) ⁇ 100%;
  • a, b, c, and d are as shown in FIG. 1 , wherein a is a cell nucleus in which no hemoglobin is stained in the nucleus and the imprinted gene is not expressed; and b is a hematoxylin staining of the cell.
  • the c is the hematoxylin staining of the cell, there are two red/brown marks in the nucleus, and the nucleus of the imprinted gene is deleted;
  • the d is the hematoxylin After staining, there are more than two red/brown markers in the nucleus, and the nuclei with abnormal copy number of the imprinted gene are imprinted.
  • the gastroscopic biopsy sample was obtained by taking a suspicious diseased tissue under a gastroscope and fixing the 10% neutral formalin solution for more than 24 hours.
  • the other detection methods were the same as those in Example 1.
  • each imprinted gene Z1 has an imprinted deletion and an increased expression level.
  • the stage of malignant potential begins to appear, and it rises rapidly to a very high level in the development of gastric cancer from early to late stage.
  • the abnormal copy number of imprinted gene Z1 begins to appear in the malignant potential stage, rises rapidly in early and middle stage gastric cancer, and rises in advanced gastric cancer stage.
  • the speed is slowed down; the imprinting of the imprinted gene Z11 begins to appear in the malignant potential stage, rapidly rises in the early stage of gastric cancer, and the rate of increase in the metastatic gastric cancer slows down, and continues to rise to a higher level in the advanced stage of gastric cancer, and the copy number of the imprinted gene Z11 is abnormal.
  • the increase of expression and expression began to appear in the malignant potential stage, and gradually increased to a higher level in the development of early and late gastric cancer; the imprinting deletion, copy number abnormality and expression increase of the imprinted gene Z16 increased rapidly in the malignant potential and early gastric cancer stage. However, there was a significant decrease in the stage of advanced gastric cancer;
  • the imprinting deletion, copy number abnormality and expression amount of the imprinted gene Z3 appear in the early stage of gastric cancer, continue to rise in the metaphase gastric cancer, and increase in the advanced gastric cancer.
  • the abnormal copy number of the imprinted gene Z4 rises rapidly in the early stage of gastric cancer, in the stage of intermediate and advanced gastric cancer.
  • the rate of increase of the immunization gene Z4 began to appear in the malignant potential stage, and it increased rapidly in the early stage of gastric cancer.
  • the stage of gastric cancer did not increase significantly, and it continued to rise in the advanced stage of gastric cancer.
  • the imprinting of the imprinted gene Z5 was in the stage of early gastric cancer. Rapidly rising, the rate of increase in the mid-stage gastric cancer phase slowed down, and the stage of advanced gastric cancer continued to rise to a higher level.
  • the abnormal copy number of the imprinted gene Z5 began to appear in the early stage of gastric cancer, rapidly increased in the metaphase gastric cancer, and increased in the advanced gastric cancer stage.
  • the expression of the imprinted gene Z5 is increased in the early stomach
  • the stage began to appear, gradually rising to a higher level in the metaphase and advanced gastric cancer; the imprinting of the imprinted gene Z6 began to appear in the early stage of gastric cancer, continued to rise in the metaphase gastric cancer, and the rate of increase in the advanced gastric cancer stage slowed down, a copy of the imprinted gene Z6
  • the number of abnormalities began to appear in the early stage of gastric cancer, and the rate of increase in the metastatic gastric cancer slowed down, and the stage of advanced gastric cancer continued to rise.
  • imprinted gene Z6 began to appear in the early stage of gastric cancer, and slowly increased in the middle and late stage gastric cancer; imprinted genes Z8 imprinting loss and expression increase began to appear in early gastric cancer, and slowly increased in metaphase gastric cancer. The rate of increase in advanced gastric cancer increased, but the level was still not high. The abnormal copy number of imprinted gene Z8 increased rapidly in early gastric cancer.
  • the imprinting deletion, copy number abnormality and expression increase of the imprinted gene Z10 began to appear in the malignant potential stage, and gradually increased to a higher level in the development of early to late gastric cancer; imprinted gene Z13's imprinted deletion and increased expression
  • the stage of malignant potential begins to appear, gradually rising in early and mid-stage gastric cancer, and the rate of increase in late gastric cancer is slowed down.
  • the copy number abnormality of imprinting gene Z13 begins to appear in the malignant potential stage, and the rate of rise in early gastric cancer slows down, and the rate of mid-stage gastric cancer rises. Increased, the rate of increase in advanced gastric cancer slowed down.
  • the tissues of 37 patients with gastric tumors including the gastroscopic biopsy samples (10 micrometers) were obtained in the same manner as in Example 1.
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 2.5%, or the total imprinted gene expression amount is less than 35%.
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 4-6.5%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 6.5-8%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 8%, or the imprinted gene total expression amount is greater than 65%, or a combination of at least two is IV;
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the total imprinted gene expression amount is less than 30%.
  • a combination of at least two of which is level 0 an imprinted gene deletion expression amount of 16-20%, an imprinted gene copy number abnormal expression amount of 1.5-2.5%, or an imprinted gene total expression amount of 30-40% or The combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than
  • the imprinting gene deletion expression amount is less than 15%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the imprinted gene total expression amount is less than 30%.
  • any one of the imprinted gene deletion expression amount is less than 10%, the imprinted gene copy number abnormal expression amount is less than 1%, or the total imprinted gene expression amount is less than 25%.
  • a combination of at least two of which is 0, an imprinting gene deletion expression amount of 10-15%, an imprinted gene copy number abnormal expression amount of 1-2%, or an imprinted gene total expression amount of 25-30% or
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 15-25%, the imprinted gene copy number abnormal expression amount is 2-3%, or the imprinted gene total expression amount is 30-45%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 3-5%, or the imprinted gene total expression amount is 45-55%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 5%, or the imprinted
  • the imprinting gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the imprinted gene total expression amount is less than 30%.
  • a combination of at least two of which is level 0 an imprinted gene deletion expression amount of 16-20%, an imprinted gene copy number abnormal expression amount of 1.5-2.5%, or an imprinted gene total expression amount of 30-40% or The combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 6%
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the total imprinted gene expression amount is less than 30%.
  • a combination of at least two of which is level 0 an imprinted gene deletion expression amount of 16-20%, an imprinted gene copy number abnormal expression amount of 1.5-2.5%, or an imprinted gene total expression amount of 30-40% or The combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the total imprinted gene expression amount is less than 30%.
  • a combination of at least two of which is level 0 an imprinted gene deletion expression amount of 16-20%, an imprinted gene copy number abnormal expression amount of 1.5-2.5%, or an imprinted gene total expression amount of 30-40% or The combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than
  • any one of the imprinted gene deletion expression amount is less than 10%, the imprinted gene copy number abnormal expression amount is less than 1%, or the total imprinted gene expression amount is less than 25%.
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 15-25%, the imprinted gene copy number abnormal expression amount is 2-3%, or the imprinted gene total expression amount is 30-45%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 3-5%, or the imprinted gene total expression amount is 45-55%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 5%, or the imprinted
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the total imprinted gene expression amount is less than 30%.
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 6%, or the imprinted gene total expression amount is greater than 65%, or a combination of at least two is IV;
  • any one of the imprinted gene deletion expression amount is less than 10%, the imprinted gene copy number abnormal expression amount is less than 1%, or the total imprinted gene expression amount is less than 25%.
  • a combination of at least two of which is 0, an imprinting gene deletion expression amount of 10-15%, an imprinted gene copy number abnormal expression amount of 1-2%, or an imprinted gene total expression amount of 25-30% or
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 15-25%, the imprinted gene copy number abnormal expression amount is 2-3%, or the imprinted gene total expression amount is 30-45%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 3-5%, or the imprinted gene total expression amount is 45-55%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 5%, or the im
  • the degree of benign and malignant diagnosis of tumors is divided into benign tumors, gastric cancer potential, early gastric cancer, metaphase gastric cancer and advanced gastric cancer:
  • the results of the diagnosis of the degree of benign and malignant tumors of the gastric tumors are that the imprinted gene Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 have an imprinted gene expression level and an abnormal expression amount of the imprinted gene copy number are less than I.
  • the imprinted gene of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13, and Z16 has an imprinted gene deletion level of I and the imprinted genes Z1, Z3, Z4, Z5
  • the abnormal expression level of the imprinted gene copy number of not more than one imprinted gene in Z6, Z8, Z10, Z11, Z13, and Z16 is any one of grade I, and is a benign tumor;
  • the result of the diagnosis of the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of I, and the imprinted gene
  • the abnormal expression level of imprinted gene copy number of at least two imprinted genes of Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 is grade I
  • imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10 is expressed in the class I and has no more than one imprinting gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13.
  • the abnormal expression level of the imprinted gene copy number is grade I and the imprinted gene deletion expression amount of the imprinted gene Z16 or the abnormal expression level of the imprinted gene copy number is grade I or above, or the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10
  • the imprinting gene with no more than one imprinted gene in Z11 and Z13 has a deletion level of II and imprinted with no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13.
  • the abnormal expression level of the gene copy number is in any of the cases of grade II, which is the potential of gastric cancer;
  • the result of the diagnosis of the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of II, imprinted gene
  • the abnormal expression level of the imprinted gene copy number of at least two imprinted genes in Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 is Grade II or imprinted genes Z1, Z3, Z4, Z5, Z6, Z8,
  • the imprinted gene deletion expression level of no more than one imprinted gene in Z10, Z11 and Z13 is grade III and no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 If the abnormal expression level of the imprinted gene copy is in any of the grade III, it is early gastric cancer;
  • the result of the diagnosis of the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of III, imprinted gene
  • the abnormal expression level of imprinted gene copy number of at least 2 imprinted genes of Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 is grade III or imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10
  • the imprinting gene with no more than one imprinted gene in Z11 and Z13 is expressed at the IV level and the imprint of no more than one imprinted gene in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 If the abnormal expression level of the gene copy number is in any of the IV grades, it is a metaphase gastric cancer;
  • the result of diagnosing the degree of benign and malignant of the gastric tumor is that the imprinted gene of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11 and Z13 has a deletion level of IV or imprinted gene Z1.
  • the abnormal expression level of the imprinted gene copy number of at least two of the Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z6, Z8, Z10, Z11 and Z13 is grade IV, and is advanced gastric cancer.
  • Tissue cell sections (10 micrometers) of esophageal tumors were obtained, and the detection method was the same as in Example 1.
  • the gastroscopic biopsy sample of the esophagus is a suspicious diseased tissue in which the esophagus is taken out under a gastroscope, and the 10% neutral formalin solution is fixed for more than 24 hours.
  • the other detection methods are the same as those in the first embodiment.
  • Fig. 8(a)-Fig. 8(j) The sensitivity of each imprinted gene to esophageal cancer is shown in Fig. 8(a)-Fig. 8(j). It can be seen from Fig. 8(a)-Fig. 8(c) that the imprinting of the imprinted gene Z1 is in the stage of malignant potential. It began to appear, and did not increase significantly in early esophageal cancer. It gradually increased to a high level in the metaphase and advanced esophageal cancer. The copy number abnormality of the imprinted gene Z1 gradually increased from the malignant potential to the advanced esophageal cancer stage, and the expression level of the imprinted gene Z1 was increased.
  • the increase is slowly rising in the development of esophageal cancer; the imprinting of the imprinted gene Z11 begins to appear in the malignant potential stage, and does not increase significantly in the early and middle esophageal cancer stages, and there is a significant increase in the late stage of esophageal cancer, a copy of the imprinted gene Z11
  • the number of abnormalities increased rapidly during the malignant potential stage, and the rate of increase in the development of early and late esophageal cancer gradually slowed down.
  • the increase in the expression of the imprinted gene Z11 did not increase significantly from malignant potential to metaphase esophageal cancer, but in advanced esophageal cancer.
  • the expression of the imprinted gene Z8 increased in the malignant potential stage, but did not increase significantly during the development of esophageal cancer; the imprinted imprinted gene Z10 was deleted in the middle esophageal cancer. The stage began to appear, and did not continue to rise in the advanced esophageal cancer stage. The abnormal copy number of the imprinted gene Z10 began to appear in the metaphase esophageal cancer stage, and continued to rise in advanced esophageal cancer, but the level was not high, and the expression level of the imprinted gene Z10 increased in malignancy.
  • the latent stage began to appear, but did not increase significantly during the development of esophageal cancer; the imprinted deletion of the imprinted gene Z13 began to appear in early esophageal cancer, gradually increased in the metaphase and advanced esophageal cancer stages, but the level was not high, and the imprinted gene Z13 was copied.
  • the number of abnormalities and increased expression increased rapidly in the mid-stage esophageal cancer stage and slowed down in the advanced esophageal cancer stage.
  • the gut biopsy samples (10 micrometers) of the esophagus were obtained from 57 patients with esophageal tumors.
  • the detection method was the same as in Example 1.
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 2.5%, or the total imprinted gene expression amount is less than 35%.
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 4-6.5%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 6.5-8%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 8%, or the imprinted gene total expression amount is greater than 65%, or a combination of at least two is IV;
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the total imprinted gene expression amount is less than 30%.
  • a combination of at least two of which is level 0 an imprinted gene deletion expression amount of 16-20%, an imprinted gene copy number abnormal expression amount of 1.5-2.5%, or an imprinted gene total expression amount of 30-40% or The combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than
  • any one of the imprinted gene deletion expression amount is less than 10%, the imprinted gene copy number abnormal expression amount is less than 1%, or the total imprinted gene expression amount is less than 25%.
  • a combination of at least two of which is 0, an imprinting gene deletion expression amount of 10-15%, an imprinted gene copy number abnormal expression amount of 1-2%, or an imprinted gene total expression amount of 25-30% or
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 15-25%, the imprinted gene copy number abnormal expression amount is 2-4%, or the imprinted gene total expression amount is 30-40%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 40-55%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 6%, or the
  • any one of the imprinted gene deletion expression amount is less than 10%, the imprinted gene copy number abnormal expression amount is less than 1%, or the total imprinted gene expression amount is less than 25%.
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 15-25%, the imprinted gene copy number abnormal expression amount is 2-3%, or the imprinted gene total expression amount is 30-45%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 3-5%, or the imprinted gene total expression amount is 45-55%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 5%, or the im
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the total imprinted gene expression amount is less than 30%.
  • a combination of at least two of which is level 0 an imprinted gene deletion expression amount of 16-20%, an imprinted gene copy number abnormal expression amount of 1.5-2.5%, or an imprinted gene total expression amount of 30-40% or The combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the total imprinted gene expression amount is less than 30%.
  • a combination of at least two of which is level 0 an imprinted gene deletion expression amount of 16-20%, an imprinted gene copy number abnormal expression amount of 1.5-2.5%, or an imprinted gene total expression amount of 30-40% or The combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the total imprinted gene expression amount is less than 30%.
  • a combination of at least two of which is level 0 an imprinted gene deletion expression amount of 16-20%, an imprinted gene copy number abnormal expression amount of 1.5-2.5%, or an imprinted gene total expression amount of 30-40% or The combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than
  • the imprinting gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 2.5%, or the imprinted gene total expression amount is less than 35%.
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than
  • any one of the imprinted gene deletion expression amount is less than 16%, the imprinted gene copy number abnormal expression amount is less than 1.5%, or the total imprinted gene expression amount is less than 30%.
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 20-25%, the imprinted gene copy number abnormal expression amount is 2.5-4%, or the imprinted gene total expression amount is 40-50%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 4-6%, or the imprinted gene total expression amount is 50-65%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 6%, or the imprinted gene total expression amount is greater than 65%, or a combination of at least two is IV;
  • any one of the imprinted gene deletion expression amount is less than 10%, the imprinted gene copy number abnormal expression amount is less than 1%, or the total imprinted gene expression amount is less than 25%.
  • the combination of at least two is class I, the imprinting gene deletion expression amount is 15-25%, the imprinted gene copy number abnormal expression amount is 2-3%, or the imprinted gene total expression amount is 30-45%, or at least The combination of the two is class II, the imprinting gene deletion expression amount is 25-30%, the imprinted gene copy number abnormal expression amount is 3-5%, or the imprinted gene total expression amount is 45-55%, or at least two
  • the combination of the species is class III, the imprinting gene deletion expression amount is greater than 30%, the imprinted gene copy number abnormal expression amount is greater than 5%, or the
  • Diagnosis of the degree of benign and malignant esophageal tumors can be divided into benign tumors, esophageal cancer potential, early esophageal cancer, meta-esophageal cancer and advanced esophageal cancer:
  • the results of the diagnosis of the degree of benign and malignant tumors of the esophageal tumor are that the imprinted gene Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 have an imprinted gene expression level and an abnormal expression amount of the imprinted gene copy number are less than I.
  • the imprinted gene of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13, and Z16 has an imprinted gene deletion level of I and the imprinted genes Z1, Z3, Z4, Z5
  • the abnormal expression level of the imprinted gene copy number of not more than one imprinted gene in Z6, Z8, Z10, Z11, Z13, and Z16 is any one of grade I, and is a benign tumor;
  • the result of the diagnosis of the degree of benign and malignant of the esophageal tumor is that the imprinted gene expression level of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is Grade I,
  • the abnormal expression level of the imprinted gene copy number of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is Grade I or imprinted genes Z1, Z3, Z4, Z5, Z6 In the Z8, Z10, Z11, Z13 and Z16, the imprinted gene deletion expression level of the imprinted gene is Grade II and the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16
  • the abnormal expression level of the imprinted gene copy number of no more than one imprinted gene is any one of the second grade, and is the esophageal cancer potential;
  • the result of the diagnosis of the degree of benign and malignant of the esophageal tumor is that the imprinted gene deletion expression level of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is II,
  • the abnormal expression level of the imprinted gene copy number of at least two imprinted genes of the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is Grade II or imprinted genes Z1, Z3, Z4, Z5, Z6 In the Z8, Z10, Z11, Z13 and Z16, the imprinted gene deletion expression level of the imprinted gene is III and the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 If the abnormal expression level of the imprinted gene copy number of no more than one imprinted gene is in any of the grade III, it is early esophageal cancer;
  • the result of the diagnosis of the degree of benign and malignant of the esophageal tumor is that the imprinted gene deletion expression level of at least two imprinted genes of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is III,
  • the abnormal expression level of imprinted gene copy number of at least two imprinted genes of imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 is grade III or imprinted genes Z1, Z3, Z4, Z5, Z6 , the imprinting gene of Z8, Z10, Z11, Z13 and Z16 is not more than one imprinted gene, and the imprinted gene is expressed in the IV level and in the imprinting genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16. If the abnormal expression level of the imprinted gene copy number of no more than one imprinted gene is any of the IV grades, it is a metaphase esophageal cancer;
  • the result of the diagnosis of the degree of benign and malignant tumors of the esophageal tumor is that the imprinted gene of at least two of the imprinted genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13 and Z16 has an imprinted gene expression level of IV or imprinted.
  • the abnormal expression level of the imprinted gene copy number of at least two of the imprinted genes in the genes Z1, Z3, Z4, Z5, Z6, Z8, Z10, Z11, Z13, and Z16 is Grade IV, and is an advanced esophageal cancer.
  • the detection model and system of the present disclosure express the impression of the imprinted defect on the sample of the esophageal and gastric tumor patients in an intuitive manner, and the method of in situ labeling of the imprinted gene is objective, intuitive, early, Accurate detection of changes in the imprinted (trace) gene and the ability to provide a quantitative model that contributes significantly to the diagnosis of esophageal and gastric tumors.

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Abstract

L'invention concerne un modèle de classification pour détecter le degré de malignité d'un néoplasme oesophagien et/ou d'un néoplasme gastrique et son utilisation. Le modèle classifie la variation dans des gènes imprimés dans des tumeurs en calculant la quantité d'expression de délétion des gènes imprimés, la quantité d'expression anormale de copies de gènes imprimés et la quantité d'expression totale des gènes imprimés. Le modèle affiche, de manière visuelle, les performances de perte d'empreintes dans des échantillons de tissus et de cellules provenant de patients atteints d'un néoplasme oesophagien et d'un néoplasme gastrique, et la détection des changements des gènes imprimés au moyen d'une méthode de marquage in situ de gènes imprimés est objective, visuelle, précoce et précise, et peut fournir un modèle quantitatif et peut contribuer au diagnostic du néoplasme oesophagien et du néoplasme gastrique.
PCT/CN2018/124373 2017-12-27 2018-12-27 Modèle de classification pour détecter le degré de malignité d'un néoplasme oesophagien et/ou d'un néoplasme gastrique et son utilisation Ceased WO2019129144A1 (fr)

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EP3822369A4 (fr) * 2018-07-13 2022-06-22 Lisen Imprinting Diagnostics, Inc. Modèle de classement utilisé pour détecter un degré de malignité et de bénignité de tumeurs pulmonaires et application correspondante

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CN1852974A (zh) * 2003-06-09 2006-10-25 密歇根大学董事会 用于治疗和诊断癌症的组合物和方法
CN1908189A (zh) * 2005-08-02 2007-02-07 博奥生物有限公司 体外辅助鉴定肠型胃癌及其分化程度的方法与专用试剂盒
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EP3822369A4 (fr) * 2018-07-13 2022-06-22 Lisen Imprinting Diagnostics, Inc. Modèle de classement utilisé pour détecter un degré de malignité et de bénignité de tumeurs pulmonaires et application correspondante

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