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Kit for genetic detection of uterine cancer

A kit and technology for uterine cancer, applied in the field of uterine cancer genetic detection kits, can solve the problems of increased risk of uterine cancer, damage, and elevated 4-hydroxyestradiol levels

Inactive Publication Date: 2009-12-23
XINBAXIANG SHANGHAI MOLECULAR MEDICAL TECH SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that the change of CYP1B1 gene Leu432 to Val432 increases the catalytic activity of the CYP1B1 gene-encoded product by 2-3 times, leading to an increase in the level of the highly carcinogenic estrogen metabolite 4-hydroxyestradiol, and 4-hydroxyestradiol It can directly induce DNA single-strand breaks, and can also generate benzoquinone metabolites through a series of redox reactions, which are electrophilic compounds that can combine with DNA molecules to form DNA adducts, causing DNA damage and leading to uterine cancer increased risk of disease

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1. Use of detection kits

[0027] Step 1: Extraction of DNA template

[0028] Genomic DNA of oral epithelial cells was extracted by silica gel adsorption.

[0029] Step 2: Real-time quantitative PCR reaction

[0030] Use the fluorescent quantitative PCR kit in the detection kit to perform 4 independent fluorescent quantitative PCR reactions. The total volume of each reaction is 10 μl, including 2 μl of DNA template with a concentration of 20 ng / μl, 1 μl of 10 X fluorescent quantitative PCR reaction buffer, 0.1μl 25mM dNTP mix, 0.6μl 25mM MgCl 2 solution, 0.025 μl (5 units / μl) Taq DNA polymerase, 0.225 μl each of 20 μM sense primer and antisense primer, 0.25 μl each of 10 μM fluorescent probe with VIC and fluorescent probe with FAM, and 5.325 μl of deionized water.

[0031] The reaction was carried out on a PCR amplification instrument, and the reaction conditions were 50° C. for 2 minutes, 95° C. for 10 minutes, and 60 cycles of 95° C. for 30 seconds and 60° ...

Embodiment 2

[0035] Example 2. Application of detection kits for individual uterine cancer genetic susceptibility testing services

[0036] Step 1: DNA Extraction

[0037] The physicians in the laboratory department of the hospital instructed the subjects to use oral swabs to sample oral epithelial cells, and the silica gel adsorption method was used to extract DNA from oral epithelial cells.

[0038] Step 2: Genotyping Assays

[0039]Using the kit provided by the present invention, the codon10 T / C SNP site on the estrogen receptor alpha gene (ER-α) of the subject's genomic DNA, the MspA1 I SNP site on the cytochrome P450 17A1 gene (CYP17A1), The MspI SNP site on the cytochrome P450 1A1 gene (CYP1A1) and the Leu432Val SNP site on the cytochrome P450 1B1 gene (CYP1B1) were detected by fluorescent quantitative PCR to determine the genotypes of these four SNP sites.

[0040] Step 3: Analysis of Individual Uterine Cancer Genetic Susceptibility

[0041] Through the analysis of the SNP genoty...

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PUM

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Abstract

The invention discloses a kit for genetic detection of uterine cancer. The kit comprises a specific primer pair and a specific probe pair which are used for detecting the mononucleotide polymorphism loci genotype of an estrogen receptor alpha gene (ER-alpha), a cytochrome P450 17A1 gene (CYP17A1), a cytochrome P450 1A1 gene (CYP1A1) and a cytochrome P450 1B1 gene (CYP1B1), a fluorescent quantitative PCR general component and the like. The kit can evaluate the genetic risk of individuals suffered from the uterine cancer by detecting the polymorphism loci genotype of the genes closely related to the genetic risk of the uterine cancer at the same time.

Description

technical field [0001] The present invention relates to the fields of molecular biology and medicine. More specifically, the present invention relates to a kit for detecting the genetic susceptibility of individual uterine cancer by simultaneously detecting the estrogen receptor α gene (ER-α) closely related to uterine cancer , Cytochrome P450 17A1 gene (CYP17A1), cytochrome P450 1A1 gene (CYP1A1), cytochrome P450 1B1 gene (CYP1B1) single nucleotide polymorphism site genotype to assess the genetic susceptibility of individual uterine cancer. Background technique [0002] Uterine cancer, also known as endometrial cancer, is a malignant tumor originating from the endometrial glands. Because it originated in the uterine body, it is also called uterine body cancer. In fertile women, the endometrium thickens every month, ready to accept a fertilized egg, and if the egg is not fertilized, the outer tissue and blood vessels of the endometrium are stripped off, and then discharged t...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N21/64
Inventor 冯哲民邹祖烨
Owner XINBAXIANG SHANGHAI MOLECULAR MEDICAL TECH SHANGHAI
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