TRAIL蛋白联合吉西他宾对肝癌细胞的体外杀伤作用

目的研究肿瘤坏死因子相关调亡诱导配体（TRAIL）和吉西他宾对肝癌细胞的杀伤作用及联合作用。方法将生长良好的肝癌细胞接种于96孔板，培养24h后，加入不同浓度TRAIL和吉西他宾，倒置显微镜下观察细胞形态变化；利用非同位素细胞增殖试剂盒和流式细胞仪分别检测细胞的生长抑制率和细胞凋亡。结果（1）TRAIL组、吉西他宾组、TRAIL和吉西他宾联合用药组24h后肝癌细胞几乎全部从培养瓶壁脱落；（2）TRAIL与吉西他宾对10株肝癌细胞均有不同程度的生长抑制作用。其中ch—hep-2对TRAIL及吉西他宾最敏感，其IC50分别为24．33ng／ml和328mg／ml；当TRAIL蛋白浓度为500ng／ml时，吉西他宾对ch—hep-2的生长抑制作用明显增强，IC50为065mg／ml，其细胞毒作用增强5倍，流式细胞仪检测两者作用后的细胞，均可检测到细胞调亡峰。结论TRAIL和吉西他宾协同诱导肝癌细胞发生调亡，TRAIL蛋白可明显增加吉西他宾的杀伤效应，它可作为临床肝癌化疗的辅助药物。

The antitumor effects of TNF-related apoptosis inducing ligand (TRAIL) combined with gemcitabin on human hepatocellular carcinoma cell line

Objective To investigate the antitumor effect of TNF-related apoptosis inducing ligand （TRAIL） combined with gemcitabin on human hepatocellular carcinoma cell lines in vitro. Methods Ten human hepatocellular carcinoma cell lines were treated with TRAIL, gemcitabin or both in vitro. Cytotoxicity was detected by using non-isotope proliferation kit. The cell morphology changes were observed using light and phase-contrast microscope. The apoptotic rates of the selected cell lines were measured by flow cytometry. Results All 10 tested cell lines were sensitive to TRAIL and gemcitabin in a dose-dependent manner, and cell line ch-hep-2 was the most sensitive one, the IC50 of which was 24.33ng/ml and 3.28ng/ml respectively. The IC50 of gemcitabin to ch-hep-2 was 0.65mg/ml with 500ng/ml TRAIL, which was one fifth of gemcitabin used alone. Flow cytometry showed an induced apoptosis peak when cells were treated by gemcitabin combined with TRAIL. Conclusion The results suggeste that TRAIL combined with low dose of gemcitabin can enhance their antitumor effects on human hepatocellular carcinoma cells.