Yang Liu-cheng, Huang Zong-hai, Kong Heng, Li Qiang, Chen Fei, Yu Jin-long, Li Zhou
Department of General Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2009 Jan;29(1):1-4.
To study the selective killing effects of adenovirus (Ad)-mediated double suicide gene system driven by KDR promoter (KDR-CdglyTK) on the human hepatic carcinoma cells and human umbilical vein endothelial cells (HUVECs).
KDR-expressing BEL-7402 and HUVECs and HepG2 cells that did not express KDR were infected by KDR-CdglyTK, and the infection efficiency and the expression of CdglyTK in the cells was detected by RT-PCR. The infected cells were treated with the the prodrugs 5-FC and GCV at different concentrations, and the cell-killing effects and bystander effects were evaluated by MTT method.
At the multiplicity of infection (MOI) of 100, the recombinant AdKDR-CDglyTK showed similar infection efficiency in the 3 cell lines. RT-PCR demonstrated CDglyTK expression in the recombinant adenovirus and the 3 infected cell lines. BEL-7402 and HUVECs infected by the KDR-CdglyTK, but not the HepG2 cells, were highly sensitive to the prodrugs (P<0.001). Bystander effects of the double suicide gene system were observed in the coculture of the infected and non-infected BEL-7402 and HUVECs.
The double suicide gene system driven by KDR promoter has specific killing effect on KDR-expressing hepatocellular carcinoma cells and HUVECs.
研究由KDR启动子驱动的腺病毒（Ad）介导的双自杀基因系统（KDR-CdglyTK）对人肝癌细胞和人脐静脉内皮细胞（HUVECs）的选择性杀伤作用。
用KDR-CdglyTK感染表达KDR的BEL-7402细胞以及不表达KDR的HUVECs和HepG2细胞，通过RT-PCR检测感染效率及细胞中CdglyTK的表达。用不同浓度的前体药物5-FC和GCV处理感染后的细胞，采用MTT法评估细胞杀伤作用及旁观者效应。
在感染复数（MOI）为100时，重组腺病毒AdKDR-CDglyTK在3种细胞系中显示出相似的感染效率。RT-PCR证实重组腺病毒及3种感染细胞系中均有CdglyTK表达。被KDR-CdglyTK感染的BEL-7402细胞和HUVECs对前体药物高度敏感，而HepG2细胞则不然（P<0.001）。在感染与未感染的BEL-7402细胞和HUVECs共培养中观察到双自杀基因系统的旁观者效应。
由KDR启动子驱动的双自杀基因系统对表达KDR的肝癌细胞和HUVECs具有特异性杀伤作用。
