苦参素逆转人肝癌细胞株HepG2/ADM多药耐药的作用  

Oxymatrine reverses multidrug resistance of humanhepatocellular carcinoma HepG2/ADM cells

作　　者：王炜[1] 黄赞松[2,3] 周喜汉[2,3] 覃月秋[2,3] 

机构地区：[1]桂林医学院,广西壮族自治区桂林市541000 [2]右江民族医学院消化疾病研究所 [3]右江民族医学院附属医院消化内科,广西壮族自治区百色市533000

出　　处：《世界华人消化杂志》2014年第10期1409-1416,共8页World Chinese Journal of Digestology

基　　金：广西自然科学基金资助项目;No.桂科自0542119;广西医疗卫生重点科研基金资助项目;No.桂卫重200887;广西中医药管理局基金资助项目;No.桂卫中gzzc0955~~

摘　　要：目的:探讨苦参素(oxymatrine,OM)体外逆转人肝癌耐药细胞株多药耐药的作用及其机制.方法:MTT法检测耐药细胞株HepG2/阿霉素(adriamycin,ADM)多药耐药性及苦参素干预前后耐药细胞药敏性的变化;流式细胞术检测苦参素对耐药细胞细胞凋亡及细胞膜表面P糖蛋白(P-glycoprotein,P-gp)表达的影响;实时荧光定量PCR法检测苦参素作用前后耐药细胞内三磷酸腺苷结合运转蛋白B1(ATP-binding cassette,sub-family B member1,ABCB1)基因的表达的变化.结果:当苦参素浓度为0.5 mg/mL时对HepG2及HepG2/ADM 2种细胞株的生长抑制率均<10%,无明显毒性作用.HepG2/ADM细胞株对ADM、顺铂(cisplatin,CDDP)、5-氟尿嘧啶(5-fluorouracil,5-FU)3种化疗药物表现出不同程度的耐药性,其耐药指数分别为27.52、5.50、4.39.无细胞毒性的0.5 mg/mL浓度的苦参素能够部分逆转HepG2/ADM对上述3种化疗药物的耐药性,其逆转倍数分别为3.68、2.82、2.03倍.苦参素作用HepG2/ADM细胞24、48、72 h后,各组细胞早期细胞凋亡率较各阴性对照组均有所提高且随OM作用时间的延长而逐渐增加(1.77%±0.25%vs 0.50%±0.10%,2.53%±0.15%vs 0.63%±0.06%,3.83%±0.42%vs 0.66%±0.06%,P<0.05).苦参素作用48 h后与阴性对照组比较HepG2/ADM耐药细胞株ABCB1基因表达下调,其比率为0.49倍;耐药细胞株细胞膜表面P-gp的表达下调(21.50%±1.08%vs 40.83%±0.35%,P<0.01).结论:苦参素能够在体外逆转人肝癌耐药细胞株HepG2/ADM的多药耐药性,其作用机制可能与其提高耐药细胞早期细胞凋亡率,下调耐药相关基因ABCB1及其蛋白表达产物P-gp的表达有关.AIM： To investigate whether oxymatrine （OM） reverses the multidrug resistance of hepatocellular carcinoma cell line HepG2/adriamycin （ADM） in vitro and to explore the potential mechanisms involved. METHODS： MTT assay was used to test the sensitivity of HepG2 cells, HepG2/ADM cells and oxymatrine-treated HepG2/ADM cells to chemotherapeutics and oxymatrine. Flow cytometry was used to detect the apoptosis and the protein levels of p-glycoprotein （P-gp） in HepG2 cells, HepG2/ADM cells and oxymatrine-treated HepG2/ADM cells. Real-time fluorescence quantitative PCR was used to determine the expression of multidrug resistance related gene ATP-binding cassette, sub-family B member 1 （ABCB1） in HepG2/ADM cells and oxymatrine- treated HepG2/ADM cells. RESULTS： Oxymatrine at concentrations lower than 0.5 mg/mL had no apparent cytotoxic effects on HepG2/ADM cells. HepG2/ADM cells showed cross-resistance to ADM, cisplatin （CDDP）, and 5-fluorouracil （5-FU）, and the resis- tance indexes were 27.52, 5.50 and 4.39, respectively. Oxymatrine could partly reverse the resis- tance of HepG2/ADM cells to ADM, CDDP and 5-FU by 3.68, 2.82 and 2.03 times, respectively. After addition of oxymatrine for 24, 48 and 72 h, the rates of early apoptosis of HepG2/ADM cells were increased gradually compared with the control group （1.77% ± 0.25% vs 0.50% ± 0.10%, 2.53% ± 0.15% vs 0.63% ± 0.06%, 3.83% ± 0.42% vs 0.66% ± 0.06%, P 〈 0.05 for all）. Oxymatrine down-regulated the expression levels of multidrug resistance-related genes ABCB1 and P-gp compared with the control group （21.50% ± 1.08% vs 40.83% ± 0.35%, P 〈 0.01）. CONCLUSION： Oxymatrine can reverse multidrug resistance of HepG2/ADM cells in vitro possibly via mechanisms related to down-regulation of ABCB1 and P-gp and promotion of cell early apoptosis.

关 键 词：苦参素 多药耐药 肝癌 逆转 

分 类 号：R735.7[医药卫生—肿瘤]