ICE基因转染联合化疗药物杀伤肝癌细胞的研究

Cytotoxicity of Hepatocellular Carcinoma Cells Induced by ICE Gene Transfetion in Combination with Antitumor Chemicals in vitro

目的 :研究人ICE基因转染联合化疗药物诱导体外杀伤肝癌细胞的作用。方法 :应用电穿孔法将构建成功含有目的基因的逆转录病毒载体 pLXSN hICE导入包装细胞系PA317,筛选G4 18抗性克隆 ,并将其病毒上清转入肝癌细胞株HepG2 ,DNA提取、电泳观察 ;采用3 H TDR掺入法观察、分析化疗药物卡铂对肝癌细胞株SMMC772 1及转入相应目的基因后的SMMC772 1 ICE ,SMMC772 1 反义hICE ,SMMC772 1 neo细胞株体外增殖的影响。结果 :ICE基因转染可诱导HepG2形成具有凋亡特征的梯状DNA ;在卡铂低浓度诱导下 ,与对照细胞相比SMMC772 1 hICE细胞株体外增殖明显受抑。结论 :人ICE基因转染可直接诱导肝癌细胞株HepG2凋亡 ,明显提高肝癌细胞SMMC772 1对化疗药物卡铂杀伤的敏感性 ,ICE基因转染联合化疗药物诱导极大增强了对肝癌细胞的杀伤作用 ,可能是治疗肝癌一个有前途的方案。

Objective: To investigate the cytotoxicity of ICE gene transfection in Combination with Antitumor Chemicals killing Hepatocellular Carcinoma Cells in vitro.Methods: The recombinant plasmid pLXSN-hICE was transferred to virus packing cell PA317 by electroporation method. And then the retrovirus containing human ICE cDNA generated by these PA317 cells were used to transfect human hepatocellular carcinoma cell line HepG2. The apoptosis of transferred cells were examined by gel electrophoresis. The influence of chemotherapeutic drug Carbo-platin to the proliferation of hepatocellular carcinoma cell line SMMC7721 and its derivative cells(SMMC7721-hICE,SMMC7721-antisence hICE,SMMC7721-neo)was observed by incorporation of 3H-TDR. Results: Electrophoresis of DNA displayed the apoptosis ladder of HepG2 transfected by ICE gene. The proliferation of SMMC7721-hICE was significantly suppressed in vitro induced by Carbo-alatin compared to the other three cell lines. Conclusion: ICE gene transfection could greatly increase the susceptibility of SMMC7721 cells to apoptotic cell death following chemotherapy. These findings suggest that combining ICE gene transfection with utilizing antitumor drugs would represent a novel approach for the effective treatment of hepatocellular carcinoma.