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为分析DNA损伤修复相关基因NBS1单核苷酸多态性(SNPs)与原发性肝癌遗传易感性的关系,并对高分辨率单链构象多态性(SSCP)检测技术在SNPs分型中的适用性进行评估,本研究对来自中国汉族人群的327例原发性肝癌以及295例阴性对照中NBS1基因常见SNPs的稀有等位基因频率进行检测和分析.此外,对NBS1基因6个常见SNPs分别选择部分样本同时进行直接序列测定,以比较2种方法的检测效果.119例原发性肝癌以及95例肝硬化/慢性肝炎组织标本的SSCP分析结果表明,6个常见NBS1基因SNPs位点(102G>A,320+208G/A,553G>C,1197T>C,2016A>G和2071-30A>T)中,SNP1197T>C的稀有等位基因频率为68.1%,显著高于肝硬化/慢性肝炎对照的57.9%(P=0.0298).对该SNP位点另外采用208份肝细胞癌和200份健康人群血液标本进一步分析,肝细胞癌SNP 1197T>C的稀有等位基因频率为66.8%,显著高于健康人群对照的58.8%(P=0.0170).其他5个SNPs的稀有等位基因频率在原发性肝癌与肝硬化/慢性肝炎之间均无显著性差异.高分辨率SSCP分析法与直接序列测定法对所选样本的SNPs基因分型结果完全一致,而且直接测序法对PCR扩增产物质量的要求相对高分辨率SSCP分析更高.研究表明,中国汉族人群NBS1基因SNP 1197T>C可能与原发性肝癌的发生相关,高分辨率SSCP技术准确度与直接测序法相当,且操作更加简便易行,非常适用于大量样本多个已知SNPs的基因分型.
In order to analyze the relationship between DNA damage repair related gene NBS1 SNPs and the genetic susceptibility to primary liver cancer, high resolution single strand conformation polymorphism (SSCP) The frequency of rare alleles of SNPs common to NBS1 in 327 primary hepatocellular carcinomas from Chinese Han population and 295 negative controls was detected and analyzed.In addition, 6 common SNPs of NBS1 gene The results of SSCP analysis of 119 cases of primary hepatocellular carcinoma and 95 cases of cirrhosis / chronic hepatitis tissues showed that the SNPs of 6 common NBS1 gene loci The rare allele frequency of SNP1197T> C was 68.1% in 102G> A, 320 + 208G / A, 553G> C, 1197T> C, 2016A> G and 2071-30A> T, significantly higher than that of cirrhosis / chronic Hepatitis control 57.9% (P = 0.0298) .There were 208 hepatocellular carcinomas and 200 healthy people in this SNP loci for further analysis, the frequency of rare allele of SNP 1197T> C in hepatocellular carcinoma was 66.8% Significantly higher than the control group of 58.8% of healthy people (P = 0.0170) .Other five SNPs rare There was no significant difference in allele frequencies between primary hepatocarcinoma and cirrhosis / chronic hepatitis.High-resolution SSCP analysis and direct sequencing showed that the SNPs genotyping results of the selected samples were identical, and direct sequencing The results of PCR-amplified product quality is higher than that of high-resolution SSCP analysis.Research shows that SNP 1197T> C of NBS1 gene in Chinese Han population may be related to the occurrence of primary liver cancer, the accuracy of high-resolution SSCP technique is similar to that of direct sequencing Fairly simple and easy to use and is well suited for genotyping multiple known SNPs in a large number of samples.