CD/5-FC系统对小鼠胃癌细胞杀伤作用的研究  

Suicide Gene Therapy for Murine Gastric Carcinoma Cell Line MFC with CD/5-FC System in Vitro

作　　者：姬舒荣[1] 刘晖[1] 张奕[2] 顾琴龙[2] 刘炳亚[2] 朱正纲[2] 林言箴[2] 

机构地区：[1]同济大学附属同济医院,200065 [2]上海第二医科大学瑞金医院,200025

出　　处：《实用癌症杂志》2002年第5期471-474,共4页The Practical Journal of Cancer

基　　金：上海市自然科学基金 (No :98ZB1 4 0 2 8);卫生部基金(No :98- 1 - 31 2 )资助

摘　　要：目的 探讨CD5-FC自杀基因系统对小鼠胃癌细胞的体外杀伤作用及其机制。方法以逆转录病毒载体介导CD基因转导小鼠胃癌细胞株MFC,采用MTT法测定CD5-FC系统的体外杀伤作用及其“旁观者”效应;利用Hoechst33258荧光染色法、琼脂糖凝胶电泳法检测凋亡细胞,同时采用半定量RT-PCR技术检测凋亡相关基因的表达。结果转导CD基因可使MFC对5-FC高度敏感,并且显示出强大的“旁观者”效应。经5-FC作用后,荧光染色可见到典型的凋亡细胞核形态学改变,琼脂糖凝胶电泳出现典型的DNA梯度,同时发现凋亡相关基因bcl-2基因表达下调,而bax与caspase-3基因表达上调。结论CD5-FC自杀基因系统对小鼠胃癌细胞MFC具有强大的杀伤作用。Objective To investigate the killing effects and its mechanisms of the CD/5 FC Enzyme/Prodrug System on murine gastric cancer cell line MFC.Methods MFC was infected by the recombinant retrovirus carrying CD gene,MTT assay was used to detect the cytotoxic effects and 'bystander effects' of the CD/5 FC system.Apoptic cells were detected by morphological changes after Hoeschst 33 258 dye staining and DNA ladder pattern,Apoptosis associated genes such as bcl 2?bax and caspase 3 were analyzed semiquantitatively with RT PCR.Results MFC cells transduced with CD gene exhibited high sensitivity to 5 FC,especially showed the pronounced 'bystander effects'.The cells stained with Hoeschst 33 258 exhibited nuclear condensation and fragmentation,mono and oligonucleosomal length DNA fragmentation 'Ladder',the hallmark of apoptosis,could be verified by agrose gel electrophoresis.Semi quantitative RT PCR analysis revealed that the expression of bax and caspase 3 genes were upregulated significantly,while the bcl 2 gene expression was downregulated.Conclusion CD/5 FC Enzyme/Prodrug system exerts a potent cytotoxic effects on the MFC,the death of MFC induced by the CD/5 FC system in vitro is mediated via apoptosis.

关 键 词：CD/5-FC系统 小鼠 胃癌 细胞杀伤作用 自杀基因系统 作用机理 胞嘧啶脱胺酶 

分 类 号：R735.2[医药卫生—肿瘤]