顺序特异性引物法对胰腺癌K-ras基因点突变的研究

Detecting K ras Gene Point Mutations in Pancreatic Adenocarcinoma by Sequence Special Primer

为研究简捷、特异、敏感的检测胰腺癌Ｋｒａｓ基因点突变的方法及其在胰腺疾病中定性诊断的价值，采用针对该基因点突变方式（ＣＧＴ、ＧＡＴ、ＧＴＴ）设计的顺序特异性引物（ＳＳＰ），先后对胰腺癌石蜡包埋组织、冰冻新鲜组织、细针穿刺组织及胰液进行多聚酶链反应（ＰＣＲ），扩增产物借助常规电泳和染色检测有无Ｋｒａｓ基因突变及突变方式。结果显示：胰腺癌石蜡包埋组织、冰冻新鲜组织、细针穿刺组织及胰液中Ｋｒａｓ基因点突变率分别为７４．２％、９５．１％、９１．４％及９４．１％，而所有被检测的慢性胰腺炎、胰岛素瘤、壶腹癌、胆管癌、十二指肠乳头癌及外伤胰腺的组织标本和胰液标本均无Ｋｒａｓ基因突变，无假阳性发生。研究表明：该检测法快速、简便、特异、敏感，具有临床实用性，可以作为鉴别胰腺肿块良。

In order to study a simple,rapid,specific and sensitive method for detecting K ras gene point mutations in pancreatic adenocarcinoma and its value to the qualitative diagnosis of pancreatic diseases.With regard to the mutant styles CGT,GTT and GAT,three kinds of sequence special primer(SSP) for ploymerase chain reaction were used to study the mutations in paraffin embedded tissues,frozen tissues,fine needle aspiration tissues and pancreatic juice of pancreatic carcinoma.The conventional gel electrophoresis and ethidium bromide were enough for detection of mutations in the amplification products.Result:K ras gene point mutation rate in above specimens were 74.2%, 95.1% ,91.4% and 94.1% respectively.No mutant K ras gene was found in tissues and pancreatic juice of all examined chronic pancreatitis,insulinoma,ampullary carcinoma,bile duct carcinoma, duodenum papillary adenocarcinoma and injured pancreas,no false positive case occurred.Conclusion:The approach is rapid,convenient,specific as well as sensitive.It may serve as a practical method for distinguishing pancreatic benign masses from malignant ones,and making a definitive diagnosis of pancreatic adenocarcinoma.