Chip-based TGCE定量检测大肠癌K-ras基因点突变  

Quantitative detection of K-ras point mutation by Chip-based TGCE

作　　者：张惠丹[1] 单连峰[2] 王晓楠[1] 任辉[3] 方瑾[1] 

机构地区：[1]中国医科大学基础医学院细胞生物学教研室,卫生部细胞生物学重点实验室,沈阳110001 [2]中国医科大学基础医学院数学教研室,沈阳110001 [3]沈阳市肛肠医院大肠外科,沈阳110002

出　　处：《解剖科学进展》2010年第4期302-306,共5页Progress of Anatomical Sciences

基　　金：国家自然科学基金资助项目(No30901727);辽宁省教育厅高等学校重点实验室项目(No20060907);辽宁省教育厅高等学校科学技术研究项目(No05L558)

摘　　要：目的建立一种定量检测大肠癌中K-ras基因突变比例的方法,并分析其与临床病理参数之间的关系。方法利用微流控温度梯度毛细管电泳(Chip-based temperature gradient electrophoresis,Chip-basedTGCE)对4例代表性大肠癌组织切片中的K-ras基因突变进行检测,通过AutoCAD软件计算出K-ras基因突变比例,结果经克隆测序校正后获得线性回归方程。应用该方法定量检测84例石蜡包埋大肠癌组织中的K-ras基因突变比例,并探讨其与临床病理参数的关系。结果 Chip-based TGCE检测K-ras基因突变比例与克隆测序检测K-ras突变比例的线性回归方程为y=0.993x-1.387,K-ras基因突变比例与肿瘤部位和浸润深度有显著相关性(P<0.05),检测获得的最低K-ras突变比例达到2.51%。结论该研究显示Chip-based TGCE是一种快速、灵敏、便捷、经济的基因突变比例定量分析方法。Objective To develop a quantitative detection method for K-ras gene point mutations in colorectal cancers and analyze the relationship between the proportion of mutant K-ras gene and clinicopathological parameters. Methods Chip-based temperature gradient capillary electrophoresis （Chip-based TGCE） method was introduced to detect K-ras gene mutations in four representative archive colorectal cancers. The proportion of mutant K-ras was obtained by AutoCAD software. After confirmed and standardized by clone sequencing, lineal regression equation was put forward, by which the proportions of mutant K-ras in 84 cases of paraffin-embedded colorectal cancer tissues was obtained. The association of the proportion of mutant K-ras gene and clinical features was evaluated simultaneously. Results The lineal regression equation of the exact proportions from clone sequencing and the non-adjusted proportions of mutant K-ras from Chip-based TGCE was y=0.993x-1.387. The proportion of mutant K-ras gene was significantly correlated with tumor location and invasive depth （P0.05）. This detection system could detect the lowest 2.51% of the mutant K-ras gene. Conclusion Chip-based TGCE is a rapid, sensitive, convenient and quantitative detection method for the proportion of mutant gene.

关 键 词：微流控芯片 温度梯度毛细管电泳 点突变检测 K-RAS基因 定量 

分 类 号：R735.34[医药卫生—肿瘤]