蟾酥活性成分下调极光激酶表达并促进肝癌细胞周期阻滞的机制研究  

Mechanism of active ingredients of Venenum Bufonis in down-regulating expression of aurora kinase and promoting cell cycle arrest in human liver cancer HepG2 cells

作　　者：王佳宝 徐忠伟[2] 王志美 代二庆[3] 徐瑞成[2,4] 

机构地区：[1]锦州医科大学中国人民武装警察部队后勤学院附属医院研究生培养基地,天津300162 [2]中国人民武装警察部队后勤学院,天津300309 [3]中国人民武装警察部队后勤学院附属医院军人医疗保健中心,天津300162 [4]天津市职业与环境危害生物标志物重点实验室,天津300309

出　　处：《中草药》2017年第18期3796-3801,共6页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81673651;81273552;81273745);武警后勤学院博士启动金项目(WHB201501)

摘　　要：目的探讨极光激酶(AURK)在肝癌中的作用,以及蟾酥活性成分华蟾毒配基和蟾蜍灵下调AURK表达和抑制肝癌Hep G2细胞增殖的机制。方法 Kaplan-Meier生存函数法分析极光激酶A(AURKA)和极光激酶B(AURKB)m RNA表达水平与肝癌患者生存期的关系;MTT法检测Hep G2细胞增殖,流式细胞术检测细胞周期变化,蛋白免疫印迹法检测AURKA、AURKB、Xklp2靶向蛋白(TPX2)、染色体结构维持蛋白2(SMC2)、DNA拓扑异构酶2(TOP2A)和细胞周期蛋白依赖性激酶1(CDK1)的表达水平。结果 Kaplan-Meier生存分析显示AURKA和AURKB m RNA表达水平与肝癌患者生存期呈现显著负相关;华蟾毒配基和蟾蜍灵可抑制Hep G2细胞生长,抑制效应呈现时间和浓度依赖性;华蟾毒配基和蟾蜍灵均引起细胞周期G2/M期阻滞,下调AURKA、AURKB、TPX2、SMC2、TOP2A和CDK1蛋白表达(P<0.05)。结论 AURKA和AURKB m RNA表达水平与患者生存期呈现显著负相关。华蟾毒配基和蟾蜍灵能有效促使AURKA和AURKB表达下调,调控有丝分裂相关分子,引起Hep G2细胞周期阻滞,从而抑制其增殖。Objective To investigate the function of aurora kinase（AURK） in liver cancer and the mechanism of cinobufagin and bufalin-induced liver cancer HepG2 cells growth inhibition by down-regulating AURK family. Methods Kaplan-Meier survival method analyzed the relationship between m RNA expression levels of AURKA and AURKB and survival periods. The viability and cell cycle of HepG2 cells were detected by MTT method and flow cytometry. Western blotting analyzed the expression levels of spindle-associated protein AURKA, AURKB, TPX2, SMC2, TOP2 A, and cyclin-dependent kinase CDK1. Results Kaplan-Meier survival analysis presented a significantly negative correlation between mR NA expression levels of AURKA and AURKB and survival periods. Cinobufagin and bufalin inhibited the growth of HepG2 cells in a time-and dose-dependent manner, and induced the cell cycle G2/M phase arrest. They all down-regulated the expression of AURKA, AURKB, TPX2, SMC2, TOP2 A, and CDK1（P〈0.05）. Conclusion There is a significantly negative correlation between m RNA expression levels of AURKA and AURKB and survivalperiods. Cinobufagin and bufalin could induce HepG2 cells growth inhibition and cell cycle arrest by down-regulating the expression of AURKA and AURKB and other mitosis-regulating proteins.

关 键 词：蟾酥 华蟾毒配基 蟾蜍灵 HEP G2细胞 极光激酶A 极光激酶B 肝癌 

分 类 号：R285.5[医药卫生—中药学]