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目的探讨HSV-TK基因/GCV系统对人膀胱癌细胞的杀伤作用.方法以潮霉素磷酸转移酶和HSV-HK的融合基因(HyTK)作为目的基因,利用阳离子脂质体将其转入膀胱癌细胞株,检测其表达及GCV对转基因细胞的杀伤作用。结果RT-PCR检测证实HyTK在转基因细胞的表达,MTT法测定示GCV对EJ/HyTK细胞有明显的杀伤作用;其IC50值为2.4mg/L,而对其亲代EJ细胞无明显毒性(P<005)。按不同比例混合EJ/HyTK细胞和亲代EJ细胞,发现GCV不仅能杀伤转入TK基因的细胞,而且对其周围未转基因的膀胱细胞存在旁观者效应。结论HSV-TK基因/GCV系统可作为膀胱癌基因治疗的一种有效方法。
Objective To investigate the killing effect of HSV-TK gene / GCV system on human bladder cancer cells. Methods The hygromycin phosphotransferase (HSV-HK) fusion gene (HyTK) was used as the target gene and transfected into bladder cancer cell lines by cationic liposomes. The expression of HSV-HSV-HSV-HK fusion gene and the killing effect of GCV on the transgenic cells were detected. Results The expression of HyTK in transgenic cells was confirmed by RT-PCR. The cytotoxicity of GCV on EJ / HyTK cells was observed by MTT assay. The IC50 value was 2.4 mg / L, but no obvious toxicity to EJ cells (P < 005). The EJ / HyTK cells and the parental EJ cells were mixed in different proportions. It was found that GCV not only killed the cells transfected with the TK gene, but also had bystander effect on the non-transgenic bladder cells around it. Conclusion The HSV-TK gene / GCV system can be an effective method for gene therapy of bladder cancer.