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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 establish a method to quantitatively detect the proportion of K-ras gene mutations in colorectal cancer, and analyze the relationship between them and clinicopathological parameters. Methods The K-ras gene mutations in four representative colorectal cancer tissue sections were detected by using Chip-based temperature gradient electrophoresis (Chip-based TGCE). K-ras gene mutations were calculated by AutoCAD software. Proportions, Results Linear regression equations were obtained after correction by cloning and sequencing. This method was used to quantitatively detect the proportion of K-ras gene mutations in 84 cases of paraffin-embedded colon cancer tissues, and to explore the relationship between K-ras gene mutations and clinicopathological parameters. Results The linear regression equation of K-ras gene mutation proportion by Chip-based TGCE and K-ras mutation ratio by clone sequencing was y=0.993x-1.387. The proportion of K-ras gene mutation was significantly correlated with the tumor site and depth of invasion ( P<0.05), the ratio of the lowest K-ras mutation obtained in the assay reached 2.51%. Conclusions This study shows that Chip-based TGCE is a rapid, sensitive, convenient, and economical method for quantitative analysis of gene mutations.