摘要
目的建立稳定、有效地检测错配修复(Mismatch Repair,MMR)基因突变的方法。方法对30例肠癌及癌旁正常组织标本分别提取DNA,设计错配修复基因hMSH1引物,应用聚合酶链式反应进行PCR扩增,10%聚丙稀酰胺凝胶电泳PCR产物,银染等技术检测分析错配修复基因hMSH1突变;hMSH1抗体为一抗,免疫组化法检测30例肠癌及例癌旁正常组织石蜡切片中hMSH1蛋白的表达。结果30例肠癌组织中发生错配修复基因hMSH1突变4例(发生率13.3%),10例癌旁正常组织中无一例发生错配修复基因hMSH1突变;免疫组化检测到30例肠癌组织中有6例hMSH1蛋白表达阴性,其中hMSH1基因突变的组织hMSH1蛋白均未表达,正常组织中hMSH1蛋白表达正常。结论PCR-SSCP银染法是检测错配修复基因突变的简捷、稳定、有效的方法之一。可成为早期诊断肿瘤发生的分子手段。在肿瘤的早期治疗中具有重要作用。
Objective To establish a stable and effective method to detect mismatch repair（MMR） in gene mutation. Methods To extract DNA from cancer and normal tissue beside cancer from 30 patients with colonic cancer,to amplify the mismatching repair gent sequence by PCR using specially designed preimers. （hMSHI）, to detect the mutation of mismatching repair gene by 10%——electrophoresis and dyeing with silver technic;to detect the expression of hMSHI protein in the cancer and normal tissue beside cancer from 30 patients with colonic cancer by the technic of immuno-biochemical.Results There is 4 （13.3%） mismatching repair gene hMSH1 happens in 30 colonic cancer patients and none in 10 normal control group.Conclusion It is a concise,stable and effective method to detect mismatching repair gone mutation by the method of PCR-SSCP. It will be a critical molecular biology method to help diagnosing colonic cancer early and treating early.
出处
《中国实验诊断学》
2006年第2期184-186,共3页
Chinese Journal of Laboratory Diagnosis
