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

Active Publication Date: 2015-07-15
SUN YAT SEN UNIV CANCER CENT
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  • Summary
  • Abstract
  • Description
  • Claims
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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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  • Applications of 3' untranslated region of BRCA2 in preparation of reagents for tumor diagnosis, treatment and prognosis
  • Applications of 3' untranslated region of BRCA2 in preparation of reagents for tumor diagnosis, treatment and prognosis
  • Applications of 3' untranslated region of BRCA2 in preparation of reagents for tumor diagnosis, treatment and prognosis

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses applications of 3' untranslated region of BRCA2 in the preparation of reagents for tumor diagnosis, treatment and prognosis. The inventor finds that the RNA expression level of BRCA2-3'UTR in a plurality of tumor cell lines increases; the miR-145 is combined with the BRCA2-3'UTR, the expression amount of miR-145 is inhibited, the expression of BRCA2-3'UTR increases, the conglomeration rate of cancer stem cells increases, the tumorigenic ability in mice is enhanced, besides, when specific locked nucleic acid (LNA) inhibits the expression and normal functions of BRCA2-3'UTR, the conglomeration rate of cancer stem cells decreases and the tumorigenic ability in mice is weakened, the results show that BRCA2-3'UTR can enhance the self-updating ability of cancer stem cells and can promote the cancer cell to be in a cancer stem cell phenotype. The expression of BRCA2-3'UTR is advantageously to be the potential target point in cancer treatment. The invention can help people to better understand the mechanism of cancer, and is capable of assisting people to evaluate the cancer risk and carry out early-stage diagnosis.

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 tumor diagnosis, treatment and prognosis reagents. Background technique [0002] The Global Cancer Epidemiological Statistics (GLOBOCAN) released by the International Agency for Research on Cancer (IARC) of the World Health Organization shows that in 2012, there were about 14.1 million new cancer cases and 8.2 million cancer deaths worldwide ( This is equivalent to 14.6% of the total death toll for the year). The most common cancers in men are lung, prostate, colorectal, and stomach; in women, breast, colorectal, cervix, and lung. Due to the cumulative increase of DNA damage with age, changes in people's lifestyles, and global population growth and aging, the global cancer rate as a whole continues to rise. It is estimated that by 2025, there will be as many as 19.3 million new cancer cases . Screenin...

Claims

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Application Information

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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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