Ad-hTERTp-单纯疱疹病毒胸苷激酶／丙氧鸟苷系统联合奥沙利铂对人类肝癌细胞的影响

Effect of Ad-hTERTp-HSV-TK/GCV system combined with oxaliplatin on human hepato carcinoma cells

目的研究腺病毒介导的Ad—hTERTp一单纯疱疹病毒胸苷激酶／丙氧鸟苷（Ad-hTERTp—HSV—TK／GCV）自杀基因系统联合奥沙利铂对人类肝癌细胞HepG2的体外杀伤作用。方法将带有hTERT启动子驱动HSV—TK基因的重组复制缺陷型腺病毒Ad-hTERTp-HSV-TK作为载体。感染复数（MOI）为100转染HepG2细胞，观察不同浓度的Ad-hTERTp—HSV-TK／GCV、奥沙利铂及二者联合对HepG2细胞生长的影响。用锥虫蓝活细胞拒染法、四甲基偶氮唑盐比色（MTF）法检测各干预组肝癌细胞生长的抑制率。结果3组HepG2细胞的生长均不同程度被抑制，且随药物浓度的增加抑制作用显著增强。Ad—hTERTp—HSV。TK／GCV联合奥沙利铂组抑制率最高（86．63％），与Ad—hTERTp—HSV．TK／GCV组抑制率（72．12％）及奥沙利铂组抑制率（59．41％）相比差异均有统计学意义（P〈0．05）。结论Ad—hTRTq：Tp—HSV—TK／GCV联合奥沙利铂可显著增强对HepG2细胞的杀伤作用，增加靶向性的同时可以降低药物浓度，对临床用药有指导意义。

Objective To evaluate the lethal effect of adenovirus-mediated Ad-hTERTp-HSV-TK/GCV suicide gene system in combination with oxaliplatin （L-OHP） on human hepato carcinoma cell line HepG2 in vitro. Methods It was used that the replication-defective adenovirus earring the HSV-TK gene under control of the hTERT promoter to transfer the HepG2 cells in vitro. Human hepato carcinoma cell line HepG2 was transfected with MOI=100. It was studied that the grow inhibitory effct with Ad-hTERTp-HSV-TK/GCV therapy, oxaliplatin, Ad-hTERTp-HSV-TK/GCV therapy in combination with oxaliplatin through different drug concentration on human hepato carcinoma cell line HepG2. The growth inhibition rate of HepG2 cells was determined by trypan blue exclusion assay and MTT. Results The growth of HepG2 cells Ad-hTERTp-HSV- TK/GCV, oxaliplatin, and combination of Ad-hTERTp-HSV-TK/GCV and oxaliplatin was significantly slower. The more concentration the greater inhibition of cell growth. The growth inhibition rate of combined Ad- hTERTp-HSV-TK/GCV with oxaliplatin was 86.63 %. The growth inhibition rate of Ad-hTERTp-HSV-TK/GCV was 72.12 % compared to the combined therapy （P =0.023）. The growth inhibition rate of oxaliplatin was 59.41% compared to the combined therapy （P =0.019）. Combination of Ad-hTERTp-HSV-TFUGCV and oxaliplatin resulted in greater inhibition of cell growth compared with TK gene and L-OHP （P 〈0.05）. Conclusion HSV-TK/GCV in combination with L-OHP can enhance the lethal effect of suicide gene therapy against HepG2 cells. It is more targetable than the function of single drug therapy. Also, it could reduce drug level and plays an important role on the future＇s clinical medication.