结直肠癌组织中K-ras基因突变的毛细管电泳检测  

Determination of K-ras gene mutation in colorectal cancer tissue by capillary electrophoresis

作　　者：石冬琴[1,2] 王荣[1,2] 谢华[1] 田薇[2] 贾正平[1] 郭建魁[1] 

机构地区：[1]全军高原损伤防治重点实验室兰州军区兰州总医院临床药理基地,甘肃兰州730050 [2]浙江农林大学林业与生物技术学院,浙江临安311300

出　　处：《色谱》2013年第6期582-586,共5页Chinese Journal of Chromatography

基　　金：国家自然科学基金项目(20775089);全军医药卫生"十一五"科研课题(06MB096)

摘　　要：通过对PCR扩增的76例结直肠癌组织及癌旁正常组织DNA基因组共152个样本纯化变性后,采用毛细管电泳-激光诱导荧光检测(CE-LIF)结合单链构象多态性(SSCP)分析方法检测了人结直肠癌组织及癌旁正常组织中K-ras基因第12/13位密码子突变。所检测的76例结直肠癌患者中有30例患者存在基因突变,并对异常片段进行测序验证,测序证实以碱基G→A点突变为主。结果表明所建立的CE-LIF技术结合SSCP分析检测K-ras基因突变的方法高效、快速、灵敏、准确,适合于临床上大样本结直肠癌中K-ras基因突变分析,对选择抗结直肠癌药物有一定的指导作用。The codons 12/13 of K-ras genomic DNA from 76 colorectal cancer tissues and normal tissues were amplified by PCR.The amplified 152 samples were purified,denatured,and then detected using capillary electrophoresis（CE）-laser induced fluorescence（LIF） with single-strand conformation polymorphism（SSCP）.The abnormal samples were further confirmed by direct-sequencing.The 30 patients of the 76 colorectal cancer patients were found gene mutation,of which the results of base G→A point mutation were confirmed by the gene sequence.To detect K-ras gene mutation that has important role in clinical forecast,diagnosis,therapy and prognosis,the CE-LIF with SSCP was applied for detecting K-ras gene.It becomes a promising tool to analyse K-ras gene mutation in the clinical diagnosis and therapy of colorectal cancer tissue.

关 键 词：毛细管电泳 K-RAS基因 单链构象多态性 基因突变 结直肠癌组织 

分 类 号：O658[理学—分析化学]