HSV-TK/GCV系统对人肺癌细胞株的杀伤效应  

The effect of herpes simplex virus thymidine kinase/ganciclovir system for human lung cancer line

作　　者：宋毅[1] 刘兆雄 黄光琦[1] 肖林[1] 

机构地区：[1]四川大学华西医院,四川成都610041 [2]德阳市药品检验所,四川德阳618000

出　　处：《华西药学杂志》2004年第3期165-167,共3页West China Journal of Pharmaceutical Sciences

摘　　要：目的　观察单纯疱疹病毒胸苷激酶基因HSV -TK/GCV系统对人肺癌细胞H4 4 6的体外杀伤作用。方法　利用脂质体(lipofectin)将重组逆转录病毒载体pLXSN -TK导入包装细胞PA317,G4 18筛选至抗性克隆出现 ,收获病毒液。加入聚凝胺感染H4 4 6细胞 ,并筛选出含TK基因的H4 4 6细胞。观察GCV对H4 4 6和不同比例混合的H4 4 6、H4 4 6 /TK的杀伤作用及旁观者效应。结果　经电镜及PCR检测证实TK基因已导入PA317、H4 4 6细胞。H4 4 6 /TK细胞对GCV的敏感性明显高于H4 4 6细胞 ;随着TK+ 细胞比例的增加 ,存活率进一步减低 ,提示GCV不仅杀死TK+ 细胞 ,也杀死混合培养的未转导TK基因的H4 4 6细胞 ,表明HSV -TK基因转导的H4 4 6细胞具有明显的旁观者效应。结论　HSV -TK/GCV系统赋予人肺癌细胞对GCV的高敏感性及旁观者效应。OBJECTIVE To study the effect of herpes simplex virus thymidine kinase/ganciclovir system killing human lung cancer cell H446.METHODS Recombinant retrovial vector-pLXSN-HSV-TK was tansduced into paking cell line PA317.Lipofectin transfection techniques,recombinant retroviral was obtained and applied to infect H446 cells.The killing effects and the bystander effect on GCV on H446/TK cell alone or a mixture of H446/TK and H446 cells in different ratio were observed.RESULTS Transduction of TK gene into PA317 and H446 cell lines were validated by PCR detection and electron microscope.The H446/TK cells were more sensitive to GCV than H446 cells.The survival rate of H446+H446/TK mixed cells decreased with the increase of H446/TK ratio in mixed cells.It showed that TK/GCV system could kill not only TK + cells but also TK - cells.The phenomenon of “bystander effect” was observed.CONCLUSION H446 cells are sensitive to HSV-TK system and have significant bystander effects.

关 键 词：HSV-TK GCV系统 肺癌细胞 杀伤效应 丙氨基鸟苷 旁观者效应 

分 类 号：R734.2[医药卫生—肿瘤]