毛细管电泳-单链构象多态分析检测胃癌中p16基因突变

p16 Gene Mutation Detection by Single Strand Conformation Polymorphism with Capillary Electrophoresis in Gastric

目的:建立快速高效检测胃癌患者胃癌组织及癌旁正常组织中p16基因突变的方法。方法:采用PCR扩增p16基因第二外显子易发生突变片段,扩增样品纯化后经95℃变性;以毛细管电泳(CE)分析法结合单链构象多态性(SSCP)对60例胃癌患者p16基因突变情况进行分析。结果:分析结果表明只有3例低分化腺癌患者存在基因突变,测序表明p16基因第二外显子碱基序列AGAC发生碱基A丢失。结论:p16基因突变可能导致胃癌的发生,但不起主导作用;CE-SSCP分析方法具有快速、灵敏、准确的特点,可用于胃癌组织中p16基因的突变分析。

Objective： To obtain high performance detection method for detecting p16 gene mutation in gastric cancer tissue and normal tissue.Methods： The part of p16 gene exon 2 which liability mutation was amplified by PCR,then amplified samples were purified and denatured at 95℃.p16 gene mutation in 55 gastric cancer was detected by capillary electrophoresis（CE）-single strand conformation polymorphism（SSCP）.Results： The results revealed that only three poorly differentiated adenocarcinoma patients have gene mutation.Further sequence analysis emonstrated that base sequence AGAC detected base A in p16 gene exon 2.Conclusion： This study showed p16 gene mutation may induce gastrric cancer,but not play an inportant role in neoplastic processes.CE-SSCP has the advantage of fast,sensitive and accurate,which can use in analysis of p16 gene mutation.