| 慢病毒介导的CD/TK基因靶向杀伤胃癌细胞的作用 |
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| 引用本文： | 孔恒,黄宗海,陈海金,陶霖玉,齐柯.慢病毒介导的CD/TK基因靶向杀伤胃癌细胞的作用[J].第四军医大学学报,2009(22). |
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| 作者姓名： | 孔恒 黄宗海 陈海金 陶霖玉 齐柯 |
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| 作者单位： | 深圳南山区人民医院甲乳外科;南方医科大学附属珠江医院普通外科; |
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| 基金项目： | 国家“863”计划项目(2001AA217171) |
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| 摘 要： | 目的:探讨慢病毒介导的血管内皮细胞生长因子受体(KDR)启动子驱动的CD/TK双自杀基因体系(FGW-KDRP-CD/TK)抑制胃癌SGC-7901细胞的体外杀伤作用.方法:用重组慢病毒FGW-KDRP-CD/TK体外感染表达KDR的SGC-7901细胞,荧光显微镜观察其感染效率;RT-PCR,WesternBlot方法检测转基因细胞CD/TK的表达;用细胞计数法绘制细胞生长曲线;用流式细胞术观察用药后细胞周期的变化.给予前药更昔洛韦(GCV)和5-氟胞嘧啶(5-FC)处理后,采用MTT法观察该体系对SGC-7901细胞杀伤效应及其旁观者效应.结果:慢病毒的感染率随病毒滴度的增高而递增.经RT-PCR,West-ernBlot检测发现转基因细胞有目的基因及蛋白产物的表达.从细胞生长曲线可以看出已转染慢病毒SGC-7901和未转染SGC-7901细胞增殖情况相似,其差异无显著性.流式细胞术检测结果与对照相比较,随着前药剂量的增大,处于S期细胞明显减少.MTT法检测结果显示前药呈剂量依赖性抑制SGC-7901细胞生长.同时该体系存在明显的旁观者效应.结论:KDR启动子可以调控融合基因体系选择性地杀伤人胃癌SGC-...
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| 关 键 词： | 自杀基因治疗 慢病毒 KDR启动子 旁观者效应 |
Lentivirus-mediated CD/TK gene selectively kills human gastric adneocarcinoma cells |
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| KONG Heng,HUANG Zong-Hai,CHEN Hai-Jin,TAO Lin-Yu,QI Ke.Lentivirus-mediated CD/TK gene selectively kills human gastric adneocarcinoma cells[J].Journal of the Fourth Military Medical University,2009(22). |
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| Authors: | KONG Heng HUANG Zong-Hai CHEN Hai-Jin TAO Lin-Yu QI Ke |
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| Affiliation: | KONG Heng1,HUANG Zong-Hai2,CHEN Hai-Jin2,TAO Lin-Yu1,QI Ke1 1Department of Breast and Thyroid Surgery,Sixth Hospital of Shenzhen,Shenzhen 518052,China,2Department of General Surgery,Zhujiang Hospital,Southern Medical University,Guangzhou 510282 |
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| Abstract: | AIM:To study the killing effect of lentivirus-mediated CD/TK double gene controlled by kinase insert domain-containing receptor(KDR)promoter on human gastric carcinoma cells SGC-7901 in vitro.METHODS:SGC-7901 cells(with KDR expression)were transfected with FGW-KDRP-CD/TK vector in vitro.The infection efficiency in SGC-7901 cells was observed under fluorescence microscope.And the expression of CD/TK in genetically modified cells was detected by RT-PCR and Western Blot.The cell growth curve was drawn accordin... |
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| Keywords: | suicide gene therapy lentivirus KDR promoter bystander effect |
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