热疗联合阿霉素和TRAIL对肝癌HepG2细胞凋亡的影响

目的:观察热疗（hyperthermia，HT）联合亚浓度阿霉素(adriamycin,ADM)和肿瘤坏死因子相关凋亡诱导配体（tumor necrosis factor related apoptosis inducing ligand,TRAIL）对肝癌HepG2细胞凋亡的影响，以确定联合方案是否具有协同作用。方法:采用CCK8法检测细胞的活性；Hoechst 33258染色实验检测细胞的形态变化；流式细胞仪检测细胞的凋亡；Western blot实验检测细胞内蛋白的表达水平。结果:HT联合ADM和TRAIL可明显抑制肝癌HepG2细胞的活性（P＜0.05），促进细胞染色质浓缩、核碎裂。流式细胞检测结果表明，HT联合ADM和TRAIL可促进细胞的早期凋亡（P＜0.001）和晚期凋亡（P＜0.01）。Western blot实验进一步证明，HT联合ADM和TRAIL通过上调DR4、DR5、caspase-3和caspase-8蛋白的表达来促进细胞的凋亡（P＜0.05）。结论:HT联合ADM和TRAIL具有协同作用，可明显增加细胞凋亡的敏感性，促进肝癌细胞的凋亡。

The study on apoptosis of hepatoma carcinoma cell HepG2 treated by hyperthermia combined with adriamycin and TRAIL

Objective:To observe the effects of hyperthermia（HT) combined with low concentration of adriamycin（ADM) and tumor necrosis factor related apoptosis inducing ligand（TRAIL) on the apoptosis of hepatoma carcinoma cell HepG2,and to determine whether the joint programme has a synergistic effect.Methods:Cell activity was detected by CCK8.The morphological changes of cells were detected by Hoechst 33258 staining.Flow cytometer assay detected the cells apoptosis.The protein expression were detected by Western blot assay.Results:HT combined with ADM and TRAIL significantly inhibited the activity of hepatoma carcinoma cell HepG2（P＜0.05) and promoted cell chromatin condensation and nuclear fragmentation.Flow cytometer assay results showed that HT combined with ADM and TRAIL can promote cells early apoptosis（P＜0.001) and late apoptosis（P＜0.01).Western blot assay further proved that the HT combined with ADM and TRAIL through up-regulation of DR4,DR5,caspase-3 and caspase-8 protein expression to promote cell apoptosis（P＜0.05).Conclusion:HT combined with ADM and TRAIL have synergistic effect.It can significantly increase the sensitivity of apoptosis and promote apoptosis of hepatoma carcinoma cell.