用PCR-SSCP银染方法检测人原发性胃癌APC基因点突变

Detection of Point Mutation of APC Gene in Human Gastric Cancer by PCR-SSCP Silver-Staining Method

在PCR基础上建立了APC基因10个片段的SSCP检测体系,对影响SSCP的各种条件进行了探讨,发现主要的影响因素是电泳温度和凝胶浓度。用SSCP银染法检测人胃癌组织中APC基因的点突变,在20例中,5例有异常电泳条带(25%),表明在相应的DNA片段中发生了突变。5例突变均位于第15号外显子,其中2例位于第4号片段(密码子998-1141);3例位于第6号片段(密码子1260-1410)。研究结果表明,APC基因突变和胃癌的发生有关。PCR-SSCP银染法快速、简便,特别适用于对大样本基因点突变的检测。

By using PCR technique,we have found that the main factors that affected the SSCP(single strand conformation polymorphism) results were the elec-trophoretic temperature and the concentration of PAG (polyacrylamide gel). SSCP silver-staining method was used to detect point mutation of APC (adenomatous polyposis coli) gene in human gastric cancer. Of the 20 cases tested, 5 abnormal shifting of single strand DNA were found (25%) ,all of which were located in exon 15, two in No. 4 fragment (condon 998-1141) ; and 3 in No. 6 fragment (condon 1260-1410). Our results showed that the development of human gastric cancer might be associated with mutation of APC gene. PCR-SSCP silver-staining method is a simple and economic method and is particularly useful in the screening of point mutations in large number of samples.