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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。
The SSCP detection system for 10 fragments of APC gene was established on the basis of PCR. Various conditions affecting SSCP were investigated. The main influencing factors were found to be the electrophoretic temperature and gel concentration. The point mutation of APC gene in human gastric cancer tissues was detected by SSCP silver staining method. Among the 20 cases, 5 cases had abnormal electrophoresis bands (25%), indicating that mutations occurred in the corresponding DNA fragments. Five of the mutations were located in exon 15, of which 2 were located in segment 4 (codon 998-1141); 3 were located in segment 6 (codons 1260-1410). The results show that APC gene mutations are related to the occurrence of gastric cancer. The PCR-SSCP silver staining method is rapid and simple, and is particularly suitable for the detection of point mutations in large samples.