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Method and reagent kit for forecasting outbreak age of carcinoma of colon

A technology of colon cancer and kits, applied in the field of medical biotechnology and genetic diagnosis, to achieve the effect of low cost, high sensitivity and accuracy, and stable results

Inactive Publication Date: 2010-12-29
FUWAI HOSPITAL OF CARDIOVASCULAR DESEASE CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, there are no related reports on methods and reagents for predicting the onset age of colon cancer by using C53 gene polymorphism at home and abroad

Method used

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  • Method and reagent kit for forecasting outbreak age of carcinoma of colon
  • Method and reagent kit for forecasting outbreak age of carcinoma of colon
  • Method and reagent kit for forecasting outbreak age of carcinoma of colon

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: The expression level of C53 is closely related to the malignant degree of colon cancer—analysis of human colon cancer tissue microarray

[0034] 1. Experimental materials:

[0035] Human colon cancer tissue chip (cc05-11-005) was purchased from Xi'an Chaoying Biotechnology Co., Ltd., including 94 spots, each spot was from the pathological tissue of different colon cancer patients; the immunohistochemical kit was purchased from Beijing Zhongshan Biology company.

[0036] 2. Experimental method:

[0037] (1) Immunohistochemical detection of the expression level of C53 gene in human colon cancer tissues

[0038] 1. C53 antibody concentration: 1:200 dilution

[0039] 2. The negative control was replaced by primary antibody dilution;

[0040] 3. Experiment detailed staining steps:

[0041] Bake slices at 60°C for 30 minutes, routine dewaxing and hydration; antigen retrieval: high-pressure retrieval of antigens with 0.01M CB (citrate buffer) (pH6.0), cool to r...

Embodiment 2

[0050] Example 2: The level of microRNA379 in colon cancer tissue is negatively correlated with the expression level of C53 gene

[0051] 1. Experimental materials:

[0052] Human colon cancer tissue chip (cc05-11-006) was purchased from Xi'an Chaoying Biotechnology Co., Ltd., including 150 spots, and each spot was from the pathological tissue of different colon cancer patients; the microRNA379 hybridization probe was synthesized by Chaoying Company; The in situ hybridization kit was purchased from Beyond Biotechnology Research Institute, and the C53 antibody was self-made in this laboratory (same as Example 1); the immunohistochemistry kit was purchased from Beijing Zhongshan Biological Company.

[0053] 2. Experimental method:

[0054] (1) In situ hybridization detection of the expression level of microRNA379 in human colon cancer tissues

[0055] Bake slices at 80°C for 15 minutes; xylene I for 15 minutes, xylene II for 15 minutes; absolute ethanol I for 5 minutes, take o...

Embodiment 3

[0064] Example 3: Effects of different genotypes at site 1808 of the C53 gene on the binding activity of microRNA379

[0065] 1. Experimental materials:

[0066] 1. Biological materials: Human colon cancer cell line HCT116 was purchased from the Cell Center, Institute of Basic Research, Chinese Academy of Medical Sciences; pMIR-Report vector, microRNA379 precursor and control were purchased from Ambion; the internal reference pRL-CMV plasmid was purchased from Promega.

[0067] 2. Tool enzymes and common reagents: Lipofectamine 2000 (LF2000), DMEM, Opti-MEM I, and FetalBovine Serum were purchased from GIBCO BRL; Dual-lucifferase Reporter Assay Kit was purchased from Promega; EndoFree Plasmid Maxi Kit was purchased from QIAGEN; The rest of the reagents were imported or domestic analytically pure.

[0068] 3. Prepare and sterilize the following reagents according to conventional methods: phosphate buffered saline (PBS buffer), DMEM high glucose medium, LB medium, ethidium bromi...

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Abstract

The invention relates to a method for forecasting the onset age of colon cancer and a kit, which pertains to the medical biotechnology and the genetic diagnosis field. The method for forecasting colon cancer susceptibility determines the genotype of C53 gene polymorphism 1808 site (rs2737) of a subject and forecasts the colon cancer susceptibility of the subject by extracting the host cell genomeDNA. The method has the advantages that: the relevance of the C53 gene polymorphism site with the onset age of colon cancer is firstly expounded, the method for forecasting colon cancer susceptibility is provided, and the method can be used for the prevention, the auxiliary diagnosis and treatment of colon cancer, as well as the development of new drugs.

Description

technical field [0001] The present invention relates to a method and a kit for predicting the onset age of colon cancer, more specifically, predicting the possible onset age of colon cancer of a subject by measuring the human C53 gene polymorphism site 1808T / C (rs2737). It can be used for auxiliary diagnosis, treatment and new drug development of diseases, and belongs to the field of medical biotechnology and gene diagnosis. Background technique [0002] Cancer is a disease that seriously endangers human health, and it is the second killer after cardiovascular and cerebrovascular diseases. According to the report of the World Health Organization (WHO), there are approximately 9 million new cancer patients worldwide every year, and more than 5 million people die. In the past 20 years, the prevalence of cancer has more than doubled. Global cancer is still on the rise. According to the forecast of the World Health Organization experts, in the next 25 years, among the more than...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 惠汝太陈敬洲石毅于晖杨涛孙凯张禅那
Owner FUWAI HOSPITAL OF CARDIOVASCULAR DESEASE CHINESE ACAD OF MEDICAL SCI
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