32例肿瘤组织中p53基因突变的PCR-SSCP、银染色检测

目的　检测肿瘤组织中 p5 3基因的突变率 ,探索其在肿瘤发展过程中的作用 .建立PCR SSCP检测 p5 3突变的常规方法 .方法　检测 32例癌组织和癌旁组织 .用盐沉淀制备样品DNA ,以p5 3基因exon7设计引物 ,用PCR SSCP结合银染色显示结果 .结果　 32例肿瘤标本检出阳性 9例 ,阳性率 2 8.1% .4例癌组织和癌旁组织均为阳性 .其中 2 2例胃癌 ,检出 4例阳性 (占 18.2 % ) ,双阳性者 2例 .结论　p5 3基因突变在肿瘤中具有普遍性 ,癌组织和癌旁组织双阳性者 ,与肿瘤的扩散有关 .该检测方法简便易行 ,有助于对 p5 3基因突变的扫描检测 .

Detection of p53 Gene Mutations in 32 Cases of Carcinoma Tissue with PCR-SSCP、 Silver Staining Method

Objective: To detect the p53 gene mutations in the human carcinoma tissue and set up a simple rapid technique for testing p53 gene mutation. Method: DNA samples were extracted from 32 cases of carcinoma tissue and near by tissues with salt and ethanol precipitation. Then the primer was designed corresponding to p53 gene exon 7, using PCR SSCP, with 10% polyacrylamide electrophoresis and silver stained to show the results. Result: 9 cases from 32 samples(28.1%) showed the positive figure, of which 4 cases showed positive both in carcinoma tissue and near by tissues. Among the 22 cases of the gastric cancer, 4 cases was positive, and 2 cases show positive both in the cancer tissue and near by tissues. Conclusion: p53 gene mutation is general in cancer, mutation figure shows both positive in cancer tissue and near by tissues is related to the infiltration of cancer. The method used as a simple screening technique for testing p53 gene mutation is a suitable method in the clinical laboratory.