腺病毒介导HSV-TK/GCV系统治疗舌癌的实验研究

目的　探讨腺病毒介导单纯疱疹病毒胸苷激酶基因 (herpessimplexvirusthymidinekinase,HSV TK) /丙氧鸟苷 (ganciclovir,GCV)自杀基因系统对舌鳞癌细胞系的治疗作用。方法　RT PCR检测重组腺病毒感染Tca8113舌鳞癌细胞系后TK基因的表达情况 ;MTT法检测HSV TK/GCV系统对舌癌细胞的杀伤作用及旁观者效应。结果　AdCMVHSV TK可在舌癌细胞中表达TK基因并具有高转染率 ;HSV TK/GCV系统对舌癌细胞具有杀伤作用 ,杀伤呈时间和剂量依赖性 ;但该系统的旁观者效应差。结论　HSV TK/GCV自杀基因系统对舌鳞癌细胞系具有一定的杀伤作用

The adenovirus-mediated HSV-TK/GCV suicide gene system in the treatment of tongue carcinoma cell line

OBJECTIVE: To study the effects of replication-defective recombinant adenovirus-mediated herpes simplex virus thymidine kinase (AdCMV HSV-TK)/ganciclovir (GCV) suicide gene system on tongue carcinoma cell line. METHODS: Replication-defective recombinant adenovirus containing CMV promoter were propagated in 293 cells, purified by cesium chloride gradient and titrated by tissue culture infectious doses50 (TCID50) methods. Infectivity test was used to determine the efficiency of adenoviral transduction into tongue carcinoma cell line (Tca8113 cell line) and RT-PCR was applied to detect the expression of TK gene after infecting Tca8113 cells. The killing effects and bystander effects of HSV-TK/GCV system on tongue carcinoma cells were detected with MTT assay. RESULTS: The adenoviral titration was about 2 x 10(10) plaque-forming units(PFU)/ml. The efficiency of adenoviral transduction into tongue carcinoma cells was more than 70 percent when multiplicity of infection (MOI) was more than 20. The recombinant adenovirus expressed TK gene after infecting Tca8113 cells in vitro. Adenovirus-mediated HSV-TK/GCV suicide gene system had killing effects on Tca8113 cells and these effects were related with time and GCV dose. The killing effects were poor after treated for 3 days and increased for 5 and 7 days. There had a good killing effect only when MOI was more than 20. When MOI was 80, GCV 5 x 10(-4) mol/L and treatment for 7 days, the viability of cells is only 4%. The bystander effect of this system on Tca8113 cells is poor. CONCLUSIONS: Only when most of the tongue carcinoma cells were transferred by AdCMV HSV-TK, HSV-TK/GCV suicide gene system has good killing effect on tongue carcinoma cell line.