阿霉素对肝癌细胞增殖及TRAIL受体表达影响的实验研究

Effect of Adriamycin on Cell Proliferation and Expression of TRAIL Receptors in Human Hepatoma Carcinoma Cells HepG2

[目的]研究亚毒性浓度阿霉素联合TRAIL蛋白对肝癌HepG2细胞增殖及TRAIL受体表达的影响。[方法]采用MTT法,分别检测阿霉素、TRAIL及亚毒性浓度阿霉素联合TRAIL对HepG2细胞的生长抑制率;RT-PCR和Westernblot分别检测阿霉素作用前后HepG2细胞TRAIL受体DR4、DR5、DcR1、DcR2mRNA和蛋白表达的水平。[结果]TRAIL对HepG2细胞增殖的抑制不存在浓度依赖性;阿霉素对HepG2细胞增殖的抑制作用存在浓度依赖性;亚毒性浓度阿霉素与亚毒性浓度TRAIL联用杀伤肿瘤的能力明显增强;无论在mRNA还是蛋白水平,阿霉素处理后HepG2细胞死亡受体DR4、DR5的表达水平显著增加,而阿霉素处理后诱骗受体DcR1、DcR2的表达量较阿霉素处理前明显减少。[结论]亚毒性浓度的阿霉素与亚毒性浓度的TRAIL联合应用具有协同作用,其机制可能是由于阿霉素在某种程度上增加了细胞表面死亡受体的表达,从而诱导了细胞凋亡的增加,说明TRAIL在肿瘤治疗方面存在潜在的应用价值。

[Purpose] To investigate the effect of subtoxic adriamycin and TRAIL on cell proliferation and expression of TRAIL receptors in human hepatoma carcinoma cells HepG2.[Methods] Growth inhibition rates were examined by MTT assay under treatment with TRAIL or adriamycin or with combination.The changes of TRAIL receptors （DR4,DR5,DcR1 and DcR2） in mRNA and protein level were quantified by RT-PCR and Western-blot analysis respectively.[Results] Adriamycin had concentration-dependent effect on growth inhibition of HepG2 cells,which was not observed in TRAIL.The killing ability was enhanced with the combination of subtoxic adriamycin and subtoxic TRAIL.Increase of TRAIL death receptors （DR4,DR5） in mRNA and protein level was detected after treating with adriamycin,however,significant decrease was observed in decoy receptors （DcR1,DcR2）.[Conclusions] Subtoxic concentration of adriamycin and TRAIL in combination presents synergistic effect,and the possible mechanism lies in that adriamycin may enhance TRAIL-induced apoptosis through upregulating death receptors,which means TRAIL has potential application value for cancer treatment.