PCR-SSP检测人胰腺癌细胞株PC-2K-ras基因点突变及其方式

目的　检测人胰腺癌细胞株PC 2K ras基因点突变及其突变方式 ,明确基因治疗靶点的碱基序列。方法　针对K ras基因第 1 2位密码子点突变方式 (CGT、CAT、GTT)设计顺序特异性引物 (SSP) ,对人胰腺癌细胞株PC 2进行聚合酶链反应 (PCR) ,扩增产物借助聚丙烯酰胺凝胶电泳判定该细胞株有无K ras基因点突变及其突变方式。结果　人胰腺癌细胞株PC 2存在K ras基因点突变 ,突变方式为CGT。结论　PCR SSP法简便快速 ,特异性高 ,本研究结果为胰腺癌的下一步基因治疗奠定了基础

Detection of K-ras gene point mutation and its style in human pancreatic carcinoma cell line PC-2 by PCR-SSP

Objective To detect the bp sequence of target position for further gene therapy the K ras gene point mutation and its mutation style in human pancreatic carcinoma cell line PC 2. Methods Three kinds of special sequence primers (SSP) for polymerase chain reaction (PCR) with regard to the mutation styles (CGT, GAT and GGT) at codon 12 of K ras were used to study the human pancreatic carcinoma cell line PC 2. The amplification products were studied with polyacrylamine gel electrophoresis to detect the point mutation and its mutation style.Results K ras gene point mutation at codon 12 was found in human pancreatic carcinoma cell line PC 2, and the mutation style was CGT. Conclusion PCR SSP is rapid, convenient, and high specific. The results provide a basis for further gene therapy for pancreatic carcinoma.