吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (01): 1-7.doi: 10.13481/j.1671-587x.20180101

• 基础研究 •    下一篇

As2O3联合γ分泌酶抑制剂MW167对人肝癌HepG2/ADM细胞耐药性的影响

杨莉1,2, 王雪雯1,2, 周明1,2, 张帆1,2   

  1. 1. 石河子大学医学院新疆地方病与民族高发病教育部重点实验室, 新疆 石河子 832002;
    2. 石河子大学医学院病理生理学教研室, 新疆 石河子 832002
  • 收稿日期:2017-05-22 出版日期:2018-01-28 发布日期:2018-01-24
  • 通讯作者: 王雪雯,教授,硕士研究生导师(Tel:0993-2058621,E-mail:263@shzu.edu.cn) E-mail:263@shzu.edu.cn
  • 作者简介:杨莉(1988-),女,河南省驻马店市人,在读医学硕士,主要从事肿瘤病理生理学方面的研究。
  • 基金资助:
    国家自然科学基金资助课题(81060174)

Effect of arsenic trioxide combined with γ-secretase inhibitor MW167 on drug resistance of human hepatocellular carcinoma HepG2/ADM cells

YANG Li1,2, WANG Xuewen1,2, ZHOU Ming1,2, ZHANG Fan1,2   

  1. 1. School of Medical Sciences, Shihezi University, Key Laboratory of Xinjiang Endemic Diseases and Ethnic Diseases with High Incidences, Ministry of Education, Shihezhi 832002, China;
    2. Department of Pathophysiology, School of Medical Sciences, Shihezhi University, Shihezhi 832002, China
  • Received:2017-05-22 Online:2018-01-28 Published:2018-01-24

摘要: 目的:探讨三氧化二砷(As2O3)和γ分泌酶抑制剂MW167单独及联合作用对HepG2/ADM细胞耐药性的影响,并阐明其作用机制。方法:MTT法检测不同浓度As2O3(0.25、0.50、1.00、2.00、4.00和8.00 mg·L-1)和MW167(10、20和40 μmol·L-1)处理HepG2/ADM细胞24、48和72 h后细胞的吸光度(A)值,计算细胞增殖抑制率,确定药物作用浓度和时间;将HepG2/ADM细胞分为空白组、As2O3组、MW167组、As2O3和MW167联合组;根据MTT结果选取无细胞毒剂量的As2O3、MW167干预各组细胞48 h后,FCM法检测细胞凋亡率;RT-PCR和Western blotting法分别检测各组细胞中Notch-1、Hes-1、MDR-1(P-gp)、Bcl-2和Bax mRNA及蛋白表达水平。结果:在同一浓度时,As2O3和MW167对HepG2/ADM细胞的增殖抑制率随着作用时间的延长而升高(P<0.05或P<0.01);在同一时间点,As2O3和MW167对HepG2/ADM细胞的增殖抑制率随药物浓度的增加而升高(P<0.05);确定As2O3和MW167的无毒剂量分别为0.25 mg·L-1和10 μmol·L-1,干预时间为48 h。药物干预48 h后,与空白组比较,各干预组细胞凋亡率均升高(P<0.05),其中以联合组升高最为明显(P<0.01)。与空白组比较,各干预组细胞中Notch-1、Hes-1、MDR-1(P-gp)和Bcl-2 mRNA及蛋白表达水平明显降低(P<0.05),其中以联合组降低最为明显(P<0.01);与空白组比较,各干预组Bax mRNA和蛋白表达水平明显升高(P<0.05),其中以联合组升高最为明显(P<0.01)。结论:As2O3可逆转HepG2/ADM细胞的耐药性,其作用机制可能与抑制细胞增殖、促进细胞凋亡、下调细胞中Notch-1、Hes-1、MDR-1(P-gp)和Bcl-2基因及蛋白表达水平及上调Bax基因和蛋白表达水平有关。

关键词: 细胞凋亡, 细胞增殖, Notch信号通路, 三氧化二砷, 肝肿瘤, 多药耐药

Abstract: Objective: To investigate the influence of arsenic troxide(As2O3) and γ-secretase inhibitor MW167 used alone or in combination in the drug resistance of HepG/ADM cells,and to clarify its mechanisms. Methods: The HepG2/ADM cells were treated with different concentrations of As2O3(0.25,0.50,1.00,2.00,4.00,8.00 mg·L-1) and MW167(10,20,40 μmol·L-1) for 24,48 and 72 h,and MTT method was used to detect the optical density (A) values,then the inhibitory rates of proliferation were calculated and the drug concentration and treatment time were confirmed.The HepG2/ADM cells were divided into blank group,As2O3 group, MW167 group and combination group; the non-cytotoxic doses of As2O3 and MW167 according to the results of MTT were chosen and the HepG2/ADM cells were intervened for 48 h; the apoptotic rates were detected by FCM; RT-PCR and Western blotting methods were employed to detect the mRNA and protein expression levels of Notch-1,Hes-1,MDR-1 (P-gp),Bcl-2 and Bax in the cells in various groups. Results: At the same concentration,the inhibitory rates of proliferation of HepG2/ADM cells were enhanced with the prolongation of the treatment time of As2O3 and MW167(P<0.05 or P<0.01); at the same time point,the inhibitory rates of proliferation of As2O3 and MW167 in the HepG2/ADM cells were increased with the increasing of drug concentration (P<0.05); the non-toxic doses of As2O3 and MW167 were 0.25 mg·L-1 and 10 μmol·L-1 and the intervention time was 48 h.After treatment for 48 h,compared with blank group,the apoptotic rates in various intervention groups were increased (P<0.05),especially in combination group (P<0.01); the expression levels of Notch-1,Hes-1,MDR-1 (P-gp),Bcl-2 mRNA and protein in various intervention groups were lower than those in blank group (P<0.05),especially in combination group (P<0.01);compared with blank group,the expression levels of Bax mRNA and protein in various intervention groups were increased (P<0.05),especially in combination group (P<0.01). Conclusion: As2O3 can reverse the drug resistance of HepG2/ADM cells,its mechanism may be related to inhibition of cell proliferation,promotion of apoptosis,down-regulation of the expression levels of Notch-1,Hes-1,MDR (P-gp),Bcl-2 and up-regulation of the expression levels of Bax mRNA and protein.

Key words: multidrug resistance, apoptosis, liver neoplasms, Notch signaling pathway, cell proliferation, arsenic trioxide

中图分类号: 

  • R735.7