胰腺癌患者血浆K-ras基因突变检测的研究

Detection of K ras gene mutations in DNA extracted from the plasma of patients with pancreatic cancer

目的 检测胰腺癌患者血浆DNA中K ras基因突变 ,并探讨将之用于胰腺癌早期诊断的可能性。 方法 本研究包括胰腺癌患者 2 2例和正常对照者 2 0例。以外周静脉血为标本 ,分离血浆DNA ,采用突变富集聚合酶链反应 限制性片断长度多态性法检测K ras基因密码子 12突变。部分扩增产物行DNA顺序分析。分析血浆K ras突变与肿瘤临床病理特征的关系。 结果 77 3 %( 17/ 2 2 )的胰腺癌患者血浆DNA中含有K ras基因密码子 12突变 ,突变与肿瘤部位、大小及TNM分期无关。 2 0例正常对照者血浆中均未检测到K ras突变。 结论 胰腺癌患者血浆中存在频繁的K ras基因突变。检测血浆K

Objectives To detect mutations of the K ras codon 12 in DNA extracted from the plasma of patients with pancreatic cancer, and to explore the possibility of using this method in early diagnosis of pancreatic cancer.Methods Plasma DNA was isolated from the blood of 22 patients with pancreatic cancer and from 20 normal controls. K ras codon 12 mutations were detected by mutant enriched polymerase chain reaction restriction fragment length polymorphism technique and subsequent product sequencing. The relation of K ras mutations in plasma to clinical features in pancreatic cancer patients was analyzed. ResultsSeventeen (77.3%) of 22 patients with pancreatic cancer had a codon 12 K ras mutation in their plasma DNA. In two patients, the PCR products were sequenced and the mutations were confirmed. The occurrence of K ras mutations in the plasma DNA was not related to tumor location, tumor size, and TNM stage. No K ras mutation was detected in the plasma specimen of any of the normal controls. Conclusions K ras mutations are frequently found in the plasma DNA of patients with pancreatic cancer. Analysis of K ras mutation in the plasma DNA may be useful in the early detection of pancreatic cancer.