摘要
目的观察以人Midkine(MK)启动子调控单纯疱疹病毒胸苷激酶基因(HSV-TK)/丙氧鸟苷(GCV)自杀基因系统在体外对人肝癌细胞的杀伤效应。方法以含有人MK启动子调控的HSV-TK基因的重组腺病毒分别感染体外培养的甲胎蛋白(AFP)阳性人肝癌细胞BEL-7402和AFP阴性人肝癌细胞SMMC-7721,RT-PCR法检测HSV-TK基因在上述两种细胞中的转录表达,观察GCV对人肝癌细胞的杀伤作用。结果GCV在体外对重组腺病毒感染的BEL-7402及SMMC-7721均有明显的杀伤作用,又以前者为著。相同的病毒滴度,其杀伤作用随着GCV浓度的增高而增强。均具有旁观者效应。结论在体外表现HSV-TK基因的AFP阳性及阴性人肝癌细胞均可被人MK启动子调控的自杀基因HSV-TK杀伤。
Objective To observe and examine the killing effect of the herpes simplex virus thymidine kinase/ganciclovir （HSV-TIC/GCV） suicide gene system transferred by adenovirus on human HCC in vitro. Methods The AFP- positive human HCC cell line BEL-7402 and AFP-negative human HCC cell line SMMC-7721 were infected with the recombinant adenoviral vector containing HSV-TK gene derived by human midkine promoter. The intracellular transcription of this recombinant was detected by RT-PCR, and the effect of GCV on killing the human HCC cells was observed using microscope and MTT. Results In vitro, GCV exerts remarkable killing effect and the ＂bystander effect＂ on AFP-positive BEL-7402 as well as AFP-negative cells SMMC-7721. The killing effect was enhanced following to the GCV concentration preconditioned with same recombinant adenovirus. Conclusion Both of AFP-positive and AFP-negative human HCC cells with HSV-tk gene expressing can be killed by suicide gene HSV-TK which is controlled by human midkine promoter.
出处
《山东医药》
CAS
北大核心
2009年第20期29-32,共4页
Shandong Medical Journal
