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The application of the 3' untranslated region of brca2 in the preparation of tumor diagnostic, therapeutic and prognostic reagents

A technology of terminal untranslated regions and reagents, which is applied in the field of preparation of reagents for tumor diagnosis, treatment and prognosis, and can solve problems such as unclear related mechanisms and poor prognosis of cancer

Active Publication Date: 2018-01-19
SUN YAT SEN UNIV CANCER CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In a variety of tumors, the transcription level of BRCA2 is up-regulated, but the protein expression is inhibited, suggesting that BRCA2-3'UTR may inhibit translation, and the overexpression of BRCA2 RNA indicates that the prognosis of cancer is poor, but it is currently in tumor Mechanisms involved in growth are unclear

Method used

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  • The application of the 3' untranslated region of brca2 in the preparation of tumor diagnostic, therapeutic and prognostic reagents
  • The application of the 3' untranslated region of brca2 in the preparation of tumor diagnostic, therapeutic and prognostic reagents
  • The application of the 3' untranslated region of brca2 in the preparation of tumor diagnostic, therapeutic and prognostic reagents

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: BRCA2 is up-regulated in tumors

[0042] (1) TCGA database analysis

[0043] Methods: In order to investigate the expression of BRCA2 in tumors, biological analysis methods were applied to analyze the RNA expression data of 16 tumors in The CancerGenome Atlas (TCGA) public database, including female reproductive system cancers: Breast invasive carcinoma, BRCA), Endometrial Carcinoma (UCEC), Cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC); Male reproductive system cancer: Prostateadenocarcinoma (PRAD); Respiratory system Cancer: Lung adenocarcinoma (LUAD), Lung squamous cell carcinoma (LUSC); Digestive system cancer: Esophagealcarcinoma (ESCA), Stomach adenocarcinoma (STAD), Hepatocellular carcinoma (Liverhepatocellular) carcinoma, LIHC), pancreatic cancer (Pancreatic adenocarcinoma, PAAD), colon adenocarcinoma (COAD); endocrine system cancer: Thyroid carcinoma (THCA); urinary tract system cancer: kidney renal clear cell carcinoma (Kidney r...

Embodiment 3

[0056] Example 3: BRCA2 RNA expression level is correlated with the overall survival prognosis of various cancer patients

[0057] BRCA2 is highly expressed at RNA levels in a variety of tumors, which may be related to the occurrence and development of cancer. Therefore, the present invention further explores the relationship between the level of BRCA2 expression and the overall survival of multiple cancer patients.

[0058] Methods: Kaplan-Meier plotter, an online tool for survival and prognosis, was used to analyze the relationship between BRCA2 mRNA expression in various cancer tissues (breast cancer, ovarian cancer, lung cancer and gastric cancer) and the overall survival prognosis of patients. Log-rank test is used to compare whether the results of different experimental groups are significantly different.

[0059] A total of 1115 cases of breast cancer patients were involved in the above. Kaplan-Meier plotter automatically selected the best points of the BRCA2 high and low expr...

Embodiment 4

[0065] Example 4: BRCA2-3’UTR interacts with microRNA

[0066] Methods: Use the online tool TargetScan to predict whether the 3'untranslated region of BRCA2 (hereinafter referred to as BRCA2-3' UTR) has a target site that binds to microRNA. Then it was verified by RNA binding protein immunoprecipitation (RIP) experiment. Construct the BRCA2-3'UTR overexpression plasmid pMSCV-BRCA2-3'UTR and transfect the breast cancer cell line MDA-MB-231 to obtain the cell line MDA-MB-231-BRCA2-3 stably overexpressing 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. Collect about 5×10 separately 6 Two MDA-MB-231-BRCA2-3'UTR cells and MDA-MB-231-Vector cells were used in RIP experiments to detect the presence of microRNA in the Argonaute complex bound to BRCA2-3'UTR, and then use real-time fluorescent quantitative PCR To identify the captured microRNA, U6 is used as an internal reference f...

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Abstract

The invention discloses the application of the 3' untranslated region of BRCA2 in the preparation of tumor diagnosis, treatment and prognosis reagents. The present invention finds that the RNA expression level of BRCA2-3'UTR in various tumor cell lines is increased; and it is found that miR-145 binds to BRCA2-3'UTR, inhibits the expression of miR-145, and the expression of BRCA2-3'UTR is up-regulated, and cancer The sphere formation rate of stem cells increased and the tumorigenesis ability in mice was enhanced. In addition, when the specific locked nucleic acid LNA inhibited the expression of BRCA2-3'UTR and performed normal functions, the sphere formation rate of cancer stem cells decreased and the tumorigenesis ability in vivo was weakened. The results showed that BRCA2 ‑3'UTR can enhance the self-renewal ability of cancer stem cells and promote the cancer stem cell-like phenotype of cancer cells. The expression of BRCA2‑3'UTR is expected to become a potential target for cancer treatment, and the present invention is of great significance for better understanding the mechanism of cancer occurrence and formulating effective measures for cancer risk assessment and early detection.

Description

Technical field [0001] The present invention relates to a new application of the 3'UTR of BRCA2, in particular to the application of the 3'UTR of BRCA2 in the preparation of reagents for tumor diagnosis, treatment and prognosis. Background technique [0002] The Global Tumor Epidemiology Statistics (GLOBOCAN) released by the International Agency for Research on Cancer (IARC) of the World Health Organization shows that in 2012 there were approximately 14.1 million new cancer cases worldwide, and the number of cancer deaths reached 8.2 million (equivalent to 14.6% of the total number of deaths throughout the year). The most common cancers in men include lung cancer, prostate cancer, colorectal cancer, and gastric cancer; in women, breast cancer, colorectal cancer, cervical cancer, and lung cancer are the most common. As DNA damage accumulates with age, changes in people's lifestyles, and global population growth and aging, the global cancer rate continues to rise as a whole. It is...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68A61K48/00A61K31/7088A61P35/00
Inventor 李隽宋立兵张鑫叶丽萍吴淑刘爱斌朱金容吴阁艳
Owner SUN YAT SEN UNIV CANCER CENT
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