融合自杀基因FCU1对结肠癌细胞杀伤作用的实验研究

目的 构建含融合自杀基因FCU1的重组质粒,经脂质体转染结肠癌细胞,同时给以前体药物5-FC,观察其对结肠癌细胞的杀伤作用.方法 采用分子克隆技术,构建含融合自杀基因FCU1的真核表达质粒pEGFP-FCU1,经脂质体转染结肠癌细胞HCT116,采用MTT法检测前体药物5-FX对HCT116杀伤作用及其旁观者效应.结果 重组质粒pEGFP-FCU1经酶切鉴定,与原始目的 片段大小一致,测序与GenBank中序列一致,转染HCT116细胞后,可见荧光蛋白的表达,前体药物5-FC显示极强的杀伤作用和旁观者效应.结论 FCU1/5-FC自杀基因系统,对结肠癌细胞具有显著的杀伤作用.

Study of the Killing Effect of Fusion Suicide Gene FCU1 for Colon Cancer Cells

Objective To construct recombinant plasmid vector containing fusion suicide gene FCU1,and study the cytotoxic effect of prodrug 5-FC on the colon cancer cell,so as to provide experimental basis for further investigation. Methods Fusion suicide gene FCU1 was cloned to pEGFP-C1 containing green fluorescence protein.Recombinant plasmid pEGFP-FCU1 was transfected into colon cancer cells HCT116 with liposome-mediated method.The cytotoxic effect and bystander effect of prodrug 5-FC on transfected HCT116 cells was determined with MTT. Results The fragment obtained from the recombinant pEGFP-FCU1 by enzymatic digestion is as big as that from pCIneo-FCU1,and the sequence of FCU1 cNDA was the same as the GenBanK.5-FC has significant cytotoxic effect and bystander effect on HCT116 cells which have been transfected with pEGFP-FCU1. Conclusion FCU1/5-FC fusion suicide gene system has significant effect on colon cancer cells.