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  

Reversing effects of total glucosides of paeony on drug resistance of BEL-7402/ADM cells

作　　者：胡礼仪[1] 侯永彬[1] 余莉华[1] 米永华[1] 王科[1] 刘北忠[1] HU Liyi HOU Yongbin YU Lihua MI Yonghua WANG Ke LIU Beizhong(Yongchuan Hospital Affiliated to Chongqing Medical University, Chongqing 402160, China)

机构地区：[1]重庆医科大学附属永川医院,重庆402160

出　　处：《山东医药》2017年第10期1-4,共4页Shandong Medical Journal

基　　金：重庆医科大学附属永川医院引进人才资助项目(YJYJ201306)

摘　　要：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(IC_(50))。取亲本组、耐药组、空载组、干扰组及用药组细胞,分别采用RT-q PCR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IC_(50)分别为0.139、8.807、4.890 nmo L/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。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-7dO2/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 ICso of ADM ih each group. The mRNA and protein expression of MDR1 and histone deaeetylase 3 （ HDAC3 ） was detected by RT-qPCR and Western blotting in the above grovps. Results ICs0 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.

关 键 词：肝癌 白芍总苷 耐药 SIRNA干扰 半数抑制浓度 多药耐药基因 去乙酰化酶 

分 类 号：R735.7[医药卫生—肿瘤]