Applications of 3' untranslated region of BRCA2 in preparation of reagents for tumor diagnosis, treatment and prognosis
A technology for terminal untranslated regions and tumors, applied in the new application field of 3'UTR, which can solve the problems of poor prognosis of cancer and unclear related mechanisms
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Embodiment 1
[0041] Example 1: BRCA2 is upregulated in tumors
[0042] (1) TCGA database analysis
[0043] Methods: In order to explore the expression of BRCA2 in tumors, biological analysis methods were used to analyze the RNA expression data of 16 tumors in The Cancer Genome Atlas (TCGA) public database, including cancers of the female reproductive system: breast invasive carcinoma (Breast invasive carcinoma) , BRCA), endometrial cancer (Endometrial Carcinoma, UCEC), cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC); male reproductive system cancer: prostate cancer (Prostate adenocarcinoma, PRAD) ; Respiratory system cancer: Lung adenocarcinoma (LUAD), lung squamous cell carcinoma (Lung squamous cell carcinoma, LUSC); Digestive system cancer: Esophageal cancer (Esophageal carcinoma, ESCA), gastric adenocarcinoma (Stomach adenocarcinoma, STAD), liver Cell carcinoma (Liver hepatocellular carcinoma, LIHC), pancreatic cancer (Pancreatic adenocarcinoma, PAAD), colon...
Embodiment 3
[0056] Example 3: The RNA expression level of BRCA2 is related to the overall survival and prognosis of various cancer patients
[0057] BRCA2 is highly expressed at the RNA level in various tumors, which may be related to the occurrence and development of cancer. Therefore, the present invention further explores the relationship between the expression level of BRCA2 and the overall survival of various cancer patients.
[0058] Methods: The Kaplan-Meier plotter, an online tool for survival prognosis, was used to analyze the relationship between the expression level of BRCA2 mRNA in various cancer tissues (breast cancer, ovarian cancer, lung cancer and gastric cancer) and the overall survival prognosis of patients. The Log-rank test was used to compare whether the results of different experimental groups had significant differences.
[0059] A total of 1,115 breast cancer patients were involved in the above. Kaplan-Meier plotter automatically selected the optimal point for th...
Embodiment 4
[0065] Example 4: BRCA2-3'UTR interacts with microRNA
[0066] Methods: The online tool TargetScan was used to predict whether the 3' untranslated region of BRCA2 (hereinafter referred to as BRCA2-3'UTR) has a target site for binding to microRNA. Then it was verified by RNA binding protein immunoprecipitation (RIP) experiment. Construct the BRCA2-3'UTR overexpression plasmid pMSCV-BRCA2-3'UTR, transfect the breast cancer cell line MDA-MB-231, and obtain the cell line MDA-MB-231-BRCA2-3 that stably overexpresses BRCA2-3'UTR 'UTR, the empty vector plasmid pMSCV-Vector was transfected into the cell line MDA-MB-231 to obtain MDA-MB-231-Vector cells as a control group. Separately collect about 5 x 10 6 MDA-MB-231-BRCA2-3'UTR cells and MDA-MB-231-Vector cells were used for RIP experiments to detect the microRNA present in the Argonaute complex combined with BRCA2-3'UTR, and then by real-time fluorescent quantitative PCR The captured microRNA was identified, and U6 was used as a...
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