首页 | 本学科首页   官方微博 | 高级检索  
     

HMGB1/AMPK/m-TOR信号通路对K562细胞自噬和#br#化学治疗耐药的影响
引用本文:刘丽英,高飞,叶艳琼,陈志衡,戴云鹏,赵平,管国涛,赵明一. HMGB1/AMPK/m-TOR信号通路对K562细胞自噬和#br#化学治疗耐药的影响[J]. 中南大学学报(医学版), 2016, 41(10): 1016-1023. DOI: 10.11817/j.issn.1672-7347.2016.10.002
作者姓名:刘丽英  高飞  叶艳琼  陈志衡  戴云鹏  赵平  管国涛  赵明一
作者单位:1. 山东大学附属省立医院儿科,济南 250021;2. 广东省人民医院心外科,广州 510010;
3. 中南大学湘雅三医院儿科,长沙 410013
基金项目:国家自然科学基金(81200378,81500231);湖南省自然科学基金(2013JJ6018,2015JJ6118)。
摘    要:观察高迁移率族蛋白1(high-mobility group box 1,HMGB1)对急性髓系白血病细胞株(human leukemiacell line,K562)自噬及化学治疗(化疗)耐药的影响,并探讨其相关的分子机制。方法:体外培养K562细胞,分为化疗药物处理组、化疗药物处理对照组、HMGB1纯化蛋白预处理组、HMGB1纯化蛋白预处理对照组、HMGB1 siRNA转染组和HMGB1 siRNA转染对照组,其中化疗药物处理组又分为长春新碱(vincristine,VCR)、足叶乙甙(etoposide,VP-16)、阿糖胞苷(cytosine arabinoside,Ara-C)、阿霉素(adriamycin,ADM)和三氧化二砷(arsenic trioxide,As2O3)处理组。细胞计数试剂盒-8检测细胞活性;Western印迹检测HMGB1,微管相关蛋白1轻链3(microtubule-associate protein1light chain3,LC3),腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)和哺乳动物雷帕霉素靶点(mammalian target of rapamycin,m-TOR)磷酸化蛋白表达水平;ELISA法检测细胞上清液HMGB1蛋白含量;单丹磺酰尸胺染色和透射电镜观察细胞自噬状态。结果:与相应对照组比较,各化疗药物处理组(VCR,VP-16,Ara-C,ADM和As2O3)的细胞活性均显著降低,HMGB1蛋白表达均显著上调(均P<0.05)。与相应对照组比较,HMGB1纯化蛋白预处理组的细胞活性显著增加(P<0.05),HMGB1蛋白表达显著上调(P<0.05)。与HMGB siRNA转染对照组比较,HMGB1 siRNA转染组的细胞活性显著降低(P<0.05),HMGB1蛋白表达显著下调(P<0.05);与相应对照组比较,HMGB1纯化蛋白处理组中LC3-II表达明显增强(P<0.05),光镜下观察到自噬小体和自噬泡数量明显增加。同时,与相应对照组比较,HMGB1纯化蛋白处理组p-AMPKa的蛋白表达明显增强,而p-mTOR表达明显减弱(均P<0.05)。结论:HMGB1可能通过AMPK/m-TOR信号通路增强K562细胞自噬发挥化疗耐药性。

关 键 词:儿童白血病  高迁移率族蛋白1  自噬  化学治疗耐药  腺苷酸活化蛋白激酶  哺乳动物雷帕霉素靶
蛋白
  

Influence of HMGB1/MAPK/m-TOR signaling pathway on cell autophagy and chemotherapy resistance in K562 cells
LIU Liying,GAO Fei,YE Yanqiong,CHEN Zhiheng,DAI Yunpeng,ZHAO Ping,GUAN Guotao,ZHAO Mingyi. Influence of HMGB1/MAPK/m-TOR signaling pathway on cell autophagy and chemotherapy resistance in K562 cells[J]. Journal of Central South University. Medical sciences, 2016, 41(10): 1016-1023. DOI: 10.11817/j.issn.1672-7347.2016.10.002
Authors:LIU Liying  GAO Fei  YE Yanqiong  CHEN Zhiheng  DAI Yunpeng  ZHAO Ping  GUAN Guotao  ZHAO Mingyi
Affiliation:1. Department of Pediatrics, Shandong Provincial Hospital Affiliated to Shangdong University, Jinan 250021;
2. Department of Cardiac Surgery, Guangdong General Hospital, Guangzhou 510010;
3. Department of Pediatrics, Third Xiangya Hospital, Central South University, Changsha 410013, China
Abstract:Objective: To observe the effect of high-mobility group box 1 (HMGB1) on autophagy and chemotherapy resistance in human leukemiacell line (K562) cells, and to explore the underlying mechanisms.Methods: The K562 cells were cultured in vitro and divided into 6 groups: a chemotherapeutic group, a chemotherapeutic control group, a HMGB1 preconditioning group, a HMGB1 preconditioning control group, a HMGB1 siRNA group and a siRNA control group. The chemotherapeutic group was further divided into a vincristine (VCR) group, an etoposide (VP-16) group, a cytosine arabinoside (Ara-C) group, a adriamycin (ADM) group and a arsenic trioxide (As2O3) group. The cell activity was evaluated by cell counting kit-8. The protein levels of HMGB1, microtubule-associate protein1light chain3 (LC3), AMP-activated protein kinase (AMPK) and mammalian target of rapamycin (m-TOR) were determined by Western blotting. The level of serum HMGB1 was evaluated by enzyme-linked immunosorbent assay (ELISA). The autophagy was examined by monodansylcadaverine staining and observed under transmission electron microscopy.Results: Compared with the control group, the cell activity was significantly decreased and the level of serum HMGB1 was significantly increased in the chemotherapeutic (VCR, VP-16, Ara-C, ADM and As2O3) groups (all P<0.05). Compared with the control group, the cell activity and the level of serum HMGB1 were significantly increased in the HMGB1 preconditioning group (both P<0.05). Compared with the siRNA control group, the cell activity and the level of serum HMGB1 were significantly decreased in the HMGB1 siRNA group (both P<0.05). Compared with the control group, the expression of LC3-II and the formation of autophagic bodies were increased in the HMGB1 preconditioning group (both P<0.05), the p-AMPK expression was increased and p-mTOR expression was decreased (both P<0.05).Conclusion: HMGB1 can increase the autophagy and promote chemotherapy resistance through the pathway of AMPK/m-TOR in K562 cells.
Keywords:children leukemia  high-mobility group box 1  autophagy  chemotherapy resistance  AMP-activated protein kinase  mammalian target protein of rapamycin  
点击此处可从《中南大学学报(医学版)》浏览原始摘要信息
点击此处可从《中南大学学报(医学版)》下载全文
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号