PCR-MASA检测胰腺癌十二指肠液及腹水中K-ras基因点突变的研究

Detection of K-ras gene point mutation in duodenal secretion and in a ascites of patients with pancreatic adenocarcinoma by PCR-MASA

目的:了解胰腺癌病人十二指肠液和腹水中K-ras基因点突变检测的临床价值。方法:采用PCR-MASA(突变特异性等位基因扩增法)分别检测胰腺癌十二指肠液和腹水中K-ras基因点突变。结果:胰腺癌十二指肠液及腹水标本中K-ras基因突变率分别为17.4%(4/23)和31.6%(6/19),而所有同时被检的急、慢性胰腺炎、胰岛素瘤、壶腹癌、胆管癌、十二指肠乳头癌、胃癌及肝癌病人的十二指肠液及腹水标本中均无K-ras基因突变发现。结论:①PCR-MASA方法简捷、特异、敏感,扩增产物只需常规电泳、染色即可观察到结果,无需酶切、杂交、放射性和非放射性显影等法。②对十二指肠液及腹水标本检测K-ras基因第12位密码子有无突变,有助于判断胰腺良、恶性病变及胰腺癌的诊断。其实用价值还有待进一步验证。

Objective To evaluate the significance of detecting K-ras gene point mutation in duodenal secretion and in ascites in patients suffering from pancreatic carcinoma. Method PCR-MASA(mutant allele-specific amplification ) was used to study K-ras gene point mutation in specimens of duodenal secretion and ascites in patients with pancreatic adenocarcinoma . Result The K-ras gene point mutation rate in duodenal secretion and in ascites were 17.4% (4/23) and 31.6% (6/19) respectively in patients with pancreatic carcinoma; while it was not noted in specimens collected from those with acute pancreatitis, chronic pancreatitis, insulinoma, ampullary carcinoma, biliary duct carcinoma,duodenal papillary adenocarcinoma , stomach cancer and liver cancer . Conclusions ①This approach is rapid, convenient, sensitive and specific. Only acrylamide gel electropheresis and ethidium bromide staining are needed for studying the amplification products; more complicated procedures as restriction enzyme digestion, mutation specific oligonucleotide probe hybridization, radioisotopic and nonradioisotopic imaging were not neened. ②Detection of K-ras gene mutation in duodenal secretion and in ascites may serve as a simple and practical method for distinguishing pancreatic benign masses from malignant ones. However, its definitive value needs to be further validated.