PCR-SSP快速检测胰腺癌K-ras基因第12位密码子点突变

Rapid and simpl e detection of K-ras gene mutation at codon 12 in psncreatlc adenocarciaoma hy PCR-SSP and its implicatioo

为研究简便、快速、特异、灵敏的检测胰腺癌Ｋ－ｒａｓ基因突变和突变方式的方法及其在胰腺肿块术前定性诊断中的价值，我们采用针对Ｋ－ｒａｓ基因点突变方式（ＣＧＴ、ＧＴＴ、ＧＡＴ）设计的顺序特异性引物（ＳＳＰ），对冰冻胰腺癌组织进行ＰＣＲ，产物借助常规电泳即可知有无Ｋ－ｒａｓ基因突变及突变方式，无露酶切、杂文、放射性及非放射性显影。结果发现：３４例胰腺癌标本皆存在Ｋ－ｒａｓ基因点突变，其中８例存在两种方式突变，１３例正常胰腺组织、１７例胰腺良性疾病、７例胆管癌、５例壶腹癌及４例十二指肠乳头癌均未见Ｋ－ｒａｓ基因点突变。结果说明该法简便、快速、特异、灵敏，对于胰腺肿块术前定性诊断具有重要参考价值。

The iancidence of K-ras gene mutations at codon 12 in patient. with pancreatic adenocarcinoma was over 90 % . The PCR was perforn.ed in fruzen tis- sues of pancreatic adenocarcinoma with the xpecial se- quence primers (SSP ) designed according to the mutant styles (CCT , GTT , GAT) of K-ras gene. The cunven- tional gel electrophoresis was enough for detection of mutations in the amplification products. Restriction en- zyme digestion . specific oligonucleotide probe hybridiza- rion , radioisotopic and non-radioisotopic autography were nnt necessary . The mutation of K-ras gene at codctn 12 was found in all 34 frozen pancreatic adenocarcinoma specimens. Among 8 of the 34 samples , there were two kinds of mutatiotns. However , mutations were not found in 13 normal pancreatic tissues , 17 pancreatic benign dis- eases . 7 bile-duct carcinoznas , 5 ampullary carcinomas and 4 duodenu papillary adenocarcinomas. This method was rapid , convenicnt , spccific as well as sensit ive and may be served as a valuable method for the diagnosis and differentiation of panereatic adenocarcitnoma.