血管内皮生长因子受体启动子驱动重组腺病毒双自杀基因系统选择性杀伤大肠癌细胞

南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (05): 524-527.

血管内皮生长因子受体启动子驱动重组腺病毒双自杀基因系统选择性杀伤大肠癌细胞

林荣凯¹, 黄宗海¹, 苏国强¹, 柯志勇², 陈建雄¹, 周俊杰¹

1. 南方医科大学珠江医院普外科, 广东, 广州, 510282;
2. 南方医科大学细胞生物教研室, 广东, 广州, 510515

出版日期:2005-05-20 发布日期:2005-05-20
基金资助:
收稿日期:2005-1-25。
基金项目:国家863计划项目(2001AA217171);广东省自然科学基金重点项目(013072)
作者简介:林荣凯(1976- ),男,南方医科大学珠江医院在读硕士研究生,主治医师,电话:020-61643211,E-mail:lrk7618401@163.com

Adenovirus-mediated CDglyTK fusion gene system driven by KDR promoter selectively kills colorectal cancer cells

LIN Rong-kai¹, HUANG Zong-hai¹, SU Guo-qiang¹, KE Zhi-yong², CHEN Jian-xiong¹, ZHOU Jin-jie¹

1. 南方医科大学珠江医院普外科, 广东, 广州, 510282;
2. 南方医科大学细胞生物教研室, 广东, 广州, 510515

Online:2005-05-20 Published:2005-05-20

摘要/Abstract

摘要： 目的探讨血管内皮生长因子受体(KDR)启动子驱动重组腺病毒CDglyTK融合基因体系对结直肠癌细胞株LOVO及人脐血管内皮细胞株ECV30的选择性杀伤作用.方法将质粒pAdEasy-KDR-CDglyTK在293细胞内包装、扩增后,体外感染表达KDR的LoVo、ECV30细胞和对照组不表达KDR的LS17T细胞,并给予不同浓度的前药GCV(ganciclovir)和/或5-FC(5-fluorocytosine),观察该体系对细胞株的杀伤效应.结果制备的病毒滴度为2.0×1012pfu/ml.3种细胞的感染率相似,且感染率随腺病毒滴度的递增而增加,当MOI为200时,所有细胞株均接近100%感染.以MOI为100的重组体分别感染各细胞株,发现其对前药的敏感性不同:表达KDR的LoVo和ECV30细胞对前药具有较高的敏感性,且二者敏感性无显著差异(P>0.1);与前二者相比,LS17T细胞对前药不敏感(P<0.001).同时,CDglyTK双自杀基因的疗效优于任一单自杀基因(P<0.001).流式细胞术检测表明该体系抑制LoVo细胞DNA的合成,表现为S期细胞比率增多及G1期细胞减少(P<0.001).结论 KDR基因启动子调控的CDglyTK融合基因体系可选择性杀伤结直肠癌LoVo细胞和血管内皮细胞.

Abstract: Objective To observe the selective killing effect of adenovirus (Ad)-mediated double suicide gene driven by kinase domain-containing receptor(KDR) promoter on human colorectal cancer LoVo cells and human umbilical vein endothelial ECV304 cells.Methods The plasmid pAdEasy-KDR-CDglyTK was transfected into 293 packaging cells for amplification of the infectious Ad and used to infect the KDR-producing cells (ECV304 and LoVo) and the KDR-nonproducing cells (LS174T) respectively. The three cells were treated with the prodrugs 5-flurocytosine (5-FC) and ganciclovir (GCV) at different concentrations after infection. The killing effects of the fusion gene system on the cells were evaluated. The distribution of cell cycle was detected by flow cytometry.Results The infection rates of the recombinant Ad were similar among the 3 cells, gradually increasing with the increment of multiplicity of infection (MOI) and reaching 100% with the MOI of 200. The LoVo cells and ECV304 cells infected with Ad-KDR-CDglyTK were highly sensitive to both of the prodrugs (P>0.1), whereas the infected LS174T cells failed to exhibit similar sensitivity (P<0.001). The killing effect of CD/TK fusion gene on the target cells was much stronger than that of either suicide gene (P<0.001). The cell cycle of LoVo cells was arrested at G1 phase. Conclusion The CD/TK fusion gene system driven by KDR promoter can selectively kill KDR-expressing human colorectal cancer LoVo cells and endothelial cells.

中图分类号:

R730.59

林荣凯¹, 黄宗海¹, 苏国强¹, 柯志勇², 陈建雄¹, 周俊杰¹. 血管内皮生长因子受体启动子驱动重组腺病毒双自杀基因系统选择性杀伤大肠癌细胞[J]. 南方医科大学学报, 2005, 25(05): 524-527.

LIN Rong-kai¹, HUANG Zong-hai¹, SU Guo-qiang¹, KE Zhi-yong², CHEN Jian-xiong¹, ZHOU Jin-jie¹. Adenovirus-mediated CDglyTK fusion gene system driven by KDR promoter selectively kills colorectal cancer cells[J]. Journal of Southern Medical University, 2005, 25(05): 524-527.

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