南方医科大学学报

南方医科大学学报 ›› 2006, Vol. 26 ›› Issue (04): 402-405.

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AdKDR-CDglyTK融合基因系统对胃癌MGC-803细胞体的外杀伤作用

周广军； 黄宗海； 俞金龙； 厉周； 苏国强；   

  1. 南方医科大学珠江医院普外科； 南方医科大学珠江医院普外科 广东 广州 510282； 广东 广州 510282；
  • 出版日期:2006-04-20 发布日期:2006-04-20
  • 基金资助:
    国家863计划生物和现代农业科技研究基金项目(2001AA217171);广东省自然科学基金重点项目(013072)~~

In vitro killing effect of adenovirus-mediated fusion gene system driven by KDR promoter on gastric cancer cells

ZHOU Guang-jun, HUANG Zong-hai, YU Jin-long, LI Zhou, SU Guo-qiang Department of General Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China   

  1. 南方医科大学珠江医院普外科； 南方医科大学珠江医院普外科 广东 广州 510282； 广东 广州 510282；
  • Online:2006-04-20 Published:2006-04-20

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摘要： 目的探讨腺病毒介导血管内皮生长因子第二受体,亦即酪氨酸激酶受体(kinase domain-containing receptor,KDR) 启动子驱动的CDglyTK融合基因体系对人胃癌细胞株MGC-803的杀伤作用。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MGC-803细胞株,并给予不同浓度的前药5-FC和／或更昔洛韦,观察该体系对MGC-803细胞株的杀伤效应及其旁观者效应;并以流式细胞仪检测细胞周期的变化,电镜观察细胞的病变。结果所得病毒滴度为2．0×1012pfu／ml。MGC-803细胞株感染率随腺病毒滴度的递增而增加。融合基因的疗效优于任一单自杀基因(P<0．001)。将感染腺病毒的细胞与未感染细胞以不同比例混合培养,观察到该体系明显的旁观者效应。流式细胞术检测表明该体系抑制MGC-803细胞DNA的合成,表现为G1期细胞比率增多及S期细胞减少(P<0．001)。同时,电镜下可见MGC-803细胞有凋亡和坏死改变。结论腺病毒介导KDR启动子驱动的CDglyTK融合基因体系可以有效地杀伤人胃癌细胞。 

Abstract: Objective To evaluate the killing effect of adenovirus (Ad)-mediated double suicide gene driven by kinase domain-containing receptor (KDR) promoter on gastric cancer MGC-803 cells. Methods The 293 packaging cells were transfected by the plasmids pAdEasy-KDR-CDglyTK to generate infectious viruses. The gastric cancer MGC-803 cells were infected by the Ad followed by treatment with 5-FC and/or ganciclovir at different concentrations. The cell-killing effects were evaluated and the bystander effects analyzed after coculture of the cells without AdKDR-CDglyTK infection with the infected cells at different ratios. The cell cycle distribution was detected by flow cytometry and the pathological changes of the cells were observed by electron microscopy. Results The infection rate of the resultant recombinant Ad in the cells increased gradually with increment of the multiplicity of infection (MOI) of the Ads. The killing effect of CD/TK fusion gene on the MGC-803 cells was much stronger than that of either of the single suicide gene (P<0.001), and considerable bystander effect was observed. The Ad infection caused MGC-803 cell growth arrest at G1 phase with onset of apoptotic and necrotic morphologies of the cells as seen under electron microscope. Conclusions The CD/TK fusion gene system driven by the KDR promoter possesses effective killing effect on the KDR-expressing gastric cancer MGC-803 cells. 

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