毛细管电泳-单链构象多态分析检测胃癌中p16基因突变

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。

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

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.