白芍总苷对肝癌耐药细胞BEL-7402/ADM耐药性的逆转作用及机制探讨

摘要

Objective To observe the reversing effects of total glucosides of paeonia (TPG) on drug resistance of BEL-7402/ADM cells in hepatocellular carcinoma.Methods Human hepatocellular carcinoma ADM sensitive BEL-7402 cells (parent group) and ADM drug-resistant BEL-7402/ADM cells (drug resistance group) were cultured in vitro.BEL-7402/ADM cells were transfected with HDAC3 siRNA interference plasmid (interference group) and empty plasmid (empty plasmid group) for 24 h,respectively,and BEL-7402/ADM cells were incubated with TPG for 48 h (drug group).Different concentrations of ADM were administered to cells in the parent group,drug resistance group and drug group for 48 h respectively.Then MTT assay was used to detect the growth inhibition rates and IC50 of ADM in each group.The mRNA and protein expression of MDR1 and histone deacetylase 3 (HDAC3) was detected by RT-qPCR and Western blotting in the above grovps.Results IC50 of ADM was 0.139 nmoL/L in the parent group,8.807 nmoL/L in the drug resistance group and 4.890 nmoL/L in the drug group.Drug reverse ratio was about 1.80 times higher and the reverse transcription ratio was 45.23％ in the drug group.The relative expression of MDR1 mRNA and protein was:drug resistance group,empty plasmid group ＞ drug group ＞ interference group ＞ parent group (all P＜0.05,0.01),the relative expression of HDAC3 and protein was:drug resistance group,empty plasmid group ＞ interference group ＞ parent group,drug group (all P＜0.05,0.01).Conclusion TPG induces the down-regulated expression of MDR1 by inhibiting the HDAC3 expression,thus reversing the drug resistance of BEL-7402/ADM cells.%目的 观察白芍总苷(TPG)对肝癌耐药细胞BEL-7402/ADM耐药性的逆转作用,并探讨其作用机制.方法 体外培养人肝癌ADM敏感细胞BEL-7402(亲本组)和ADM耐药细胞BEL-7402/ADM(耐药组),将BEL-7402/ADM细胞转染HDAC3基因siRNA干扰质粒(干扰组)及空载质粒(空载组)24 h,用白芍总苷孵育BEL-7402/ADM细胞48 h(用药组).取亲本组、耐药组、用药组细胞,分别以不同浓度ADM作用48 h;MTT法测算增殖抑制率,计算各组ADM的半数抑制浓度(IC50).取亲本组、耐药组、空载组、干扰组及用药组细胞,分别采用RT-qPCR法和Western blot法检测细胞多药耐药基因MDR1和去乙酰化酶(HDAC3) mRNA和蛋白.结果 亲本组、耐药组和用药组细胞ADM的IC50分别为0.139、8.807、4.890 nmoL/L;用药组细胞对ADM耐药的逆转倍数为1.80倍,逆转率为45.23％.MDR1 mRNA及蛋白相对表达量耐药组、空载组＞用药组＞干扰组＞亲本组(P均＜0.05或0.01),HDAC3 mRNA及蛋白相对表达量耐药组、空载组＞干扰组＞亲本组、用药组(P均＜0.05或0.01).结论 白芍总苷通过抑制HDAC3的表达,引起MDR1表达下调,从而逆转BEL-7402/ADM细胞的耐药性.