慢病毒介导的双自杀基因对大肠癌细胞的靶向杀伤作用
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陈海金, 黄宗海, 苏国强, 吴爱国, 俞金龙
大肠癌;KDR启动子;自杀基因;慢病毒,陈海金,黄宗海,苏国强,吴爱国,俞金龙,陈海金,通讯作者,Targetedkillingeffectoflen
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陈海金, 黄宗海, 苏国强, 吴爱国, 俞金龙, 南方医科大学珠江医院普外科 广东省广州市 510280
陈海金, 2004年南方医科大学博士生, 主要从事大肠癌基因治疗方面研究.
广东省自然科学基金资助项目, No. 013072
通讯作者: 黄宗海, 510280, 广东省广州市, 珠江医院普外科. drhuangzh@yahoo.com.cn
电话: 020-31396133 传真: 020-61643213
收稿日期: 2006-03-12 接受日期: 2006-05-08
Targeted killing effect of lentivirus-mediated CD/TK suicide genes on colorectal carcinoma cells
Hai-Jin Chen, Zong-Hai Huang, Guo-Qiang Su, Ai-Guo Wu, Jin-Long Yu
Hai-Jin Chen, Zong-Hai Huang, Guo-Qiang Su, Ai-Guo Wu, Jin-Long Yu, Department of General Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, Guangdong Province, China
Supported by the Natural Science Foundation of Guangdong Province, No. 013072
Correspondence to: Zong-Hai Huang, Department of General Surgery, Zhujiang Hospital, Guangzhou 510280, Guangdong Province, China. drhuangzh@yahoo.com.cn
Received: 2006-03-12 Accepted: 2006-05-08
Abstract
AIM: To investigate the killing effect of lentivirus-mediated double suicide genes under the regulation of kinase domain-containing receptor (KDR) promoter targeted on colorectal carcinoma cells.
METHODS: 293FT packaging cells were transfected by the constructed plasmids pLenti6/V5-D-KDR-CDglyTK and pLenti6/V5-D-GFP. After blasticidin selection and cell cloning, the infectious viruses were generated. Then SW620 (with KDR expression) and LS174T cells (without KDR expression) were transfected with the obtained viruses by lipofectamin 2000. The transfection efficacy was evaluated by the fluorecence microscopy. The expression of CDglyTK was detected by reverse transcription-polymerase chain reaction (RT-PCR). After treatment with 5-FC and GCV, the killing effects and bystander effect of CD/TK suicide genes on the two kinds of cell lines were assessed.
RESULTS: The transfection efficacy was not significantly different between SW620 and LS174T cells, and elevated with the increase of virus titer ......
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