HyTK基因转导联合丙氧鸟苷对鼠膀胱癌细胞的体外杀伤作用

目的　探索HyTK基因转导丙氧鸟苷 (ganciclovir,GCV)对鼠膀胱癌细胞的影响。方法　脂质体介导法将PL(HyTK)SN质粒转染PA3 17细胞 ,潮霉素B筛选 ,直到出现抗性克隆 ,扩增培养 ,NIH3T3细胞测定病毒滴度 ,用逆转录病毒上清液感染T73 9细胞 ,潮霉素B筛选 ,直到出现抗性克隆 ,扩增培养 ,命名为T73 9 TK。RT PCR对T73 9 TK细胞进行鉴定。然后观察其在体外膀胱癌细胞的杀伤作用。结果　RT PCR证实HyTK基因已插入T73 9细胞并有其mRNA的表达 ,经不同浓度GCV处理后的T73 9 TK细胞均表现出有杀伤作用 ,而T73 9细胞在各个剂量的GCV作用下生长均良好。结论　GCV对转导了HyTK基因的T73 9细胞有显著杀伤作用

The killing effects of ganciclovir on HyTK gene-transduced murine bladder cancer cell line T739 cells in vitro

Objective To investigate the effects of the HyTK gene transduction on murine bladder carcinoma cells in vitro . Methods The retroviral vector plasmid PL(HyTK)SN was transfected into packaging cell line PA317 using the liposome method. The obtained recombinant retroviral supernatant fluid was used to infect murine bladder cancer cell line T739 cells. B resistant colonies, named as T739 TK, were obtained after hygromycin B selection. The integration and expression of HyTK gene in T739 TK cells were identified by RT PCR. The in vitro killing effects of GCV on T739 TK cells were observed. Results RT PCR showed that HyTK gene was inserted into T739 cells successfully and there was mRNA expression of HyTK gene. In vitro , T739 TK cells expressed the killing effects after treatment with GCV of different concentrations. However, T739 cells grew well at different doses of GCV. Conclusion GCV has obvious killing effect on HyTK gene transfected T739 cells in vitro .