【摘要】 目的:探讨抗凋亡蛋白Mcl-1在GCDA诱导的肝癌细胞耐药中的作用及其机制。方法:培养3种肝癌细胞系,用免疫荧光法和Western blot技术检测Mcl-1的表达;GCDA±CYC处理HepG2细胞,采用Western blot技术检测Mcl-1的半衰期变化;用抗癌药物Irinotecan与GCDA对HepG2细胞进行处理,采用MTT法和Western blot技术分别检测细胞增殖抑制率和Mcl-1的表达变化;用RNA干扰技术下调Mcl-1,检测化疗药物对HepG2细胞的敏感性。结果:Mcl-1在肝癌细胞中广泛表达;GCDA能延长Mcl-1的半衰期至6h以上,并明显减弱化疗药物对抗凋亡蛋白Mcl-1的抑制作用,降低癌细胞的药物敏感性;RNA干扰下调Mcl-1能增加癌细胞的药物敏感性。结论:胆盐(GCDA)能诱导HepG2细胞产生耐药性,其作用机制可能是通过延长Mcl-1半衰期增加其蛋白稳定性和抗凋亡作用来促使肝癌细胞抗药的。更多还原

【Abstract】 Objective: To study the role of anti-apoptotic protein Mcl-1 in bile salt(GCDA)-induced chemoresistance of hepatocellular carcinoma cells.Methods: Three HCC cell lines were cultured in CO2 incubator.Expression of Mcl-1 was analyzed by immunofluorescence and Western blot.HepG2 cells were treated by GCDA±CYC or Irinotecan±GCDA.Protein expression and cell viability were assessed by Western blot and MTT,respectively.Drug sensitivity was detected after down-regulation of Mcl-1 by RNA interference in HepG2 cells.Results: Mcl-1 protein is extensively expressed in HCC cells.GCDA prolongs half-life of Mcl-1 from 2-3h to more than 6h and reduces inhibitory action of Irinotecan to Mcl-1.Down-regulation of Mcl-1 by RNA interference can enhance drug sensitivity of HepG2 cells.Conclusion: Bile salt(GCDA) can induce chemoresistance of HepG2 cells.This occurs by increasing protein stability and anti-apoptotic activity of Mcl-1.更多还原