融合型自杀基因Fcy∶∶Fur/5-FC对人卵巢癌细胞杀伤作用的研究  

Killing effects of Fcy∶∶Fur/5-FC transfection on human ovarian cancer cells in vitro

作　　者：伍霞[1] 钟玲[1] 池余刚[1] 蒋兴伟[1] 王勇[1] 

机构地区：[1]重庆医科大学附属第二医院妇产科,重庆400010

出　　处：《第三军医大学学报》2008年第23期2226-2228,共3页Journal of Third Military Medical University

摘　　要：目的研究融合型自杀基因Fcy∶∶Fur联合5-氟胞嘧啶(5-FC)对人卵巢癌细胞株(SKOV3)的杀伤作用。方法构建含有融合型自杀基因Fcy∶∶Fur的荧光真核表达质粒,转染SKOV3细胞,筛选稳定表达的细胞株,RT-PCR和Western blot检测Fcy∶∶Fur表达。以MTT法和FCM法检测5-FC对Fcy∶∶Fur转基因细胞的杀伤效应。结果测序鉴定证实插入片段正确。细胞转染24h后,60%转染细胞发出绿色荧光。经G418筛选,RT-PCR和Westernblot法检测到Fcy∶∶Fur表达。加入5-FC后,MTT法检测到明显杀伤效应,FCM法检测到显著凋亡峰。结论融合型自杀基因Fcy∶∶Fur联合5-FC对人卵巢癌细胞SKOV3有明显的杀伤作用。Objective To investigate the killing effects of Fcy：： Fur suicide gene therapy system on human ovarian cancer cells （SKOV3）. Methods The EGFP（ enhanced green fluorescent protein）-labeled eukaryotic expression plasmid encoding Fcy：： Fur suicide gene fragment was constructed. The vector was transferred into SKOV3 cells. The stable transfectants were screened and analyzed by RT-PCR and Western blot analysis for the target gene expression. The killing efficacy of 5-FC on the transfected cells was determined by MTT assays and flow cytometry. Results Correct construction of pEGFP-N1-Fcy：： Fur was identified by nucleotide sequencing. About 60% transfected cells displayed green fluorescence after 24 hours＇ transfection. RT-PCR and Western blotting indicated that the transfected cells expressed Fcy：： Fur gene. MTT and flow cytometry showed 5-FC kill- ed transfected SKOV3 cells. Conclusion Fusion suicide gene therapy system Fcy：： Fur/5-FC has a significant efficacy to kill human ovarian cancer cell line SKOV3.

关 键 词：自杀基因 Fcy∶∶Fur 基因转染 稳定表达 SKOV3 

分 类 号：R394-33[医药卫生—医学遗传学] R73-362[医药卫生—基础医学]