摘要
目的研究去甲泽拉木醛(demethylzeylasteral,ZST93)在体外抑制慢性髓系白血病(chronic myeloid leukemia,CML)细胞的增殖作用,并初步探讨其可能的作用机制。方法以K562细胞为研究对象,采用CCK-8、细胞生长曲线和倒置显微镜检测ZST93对K562细胞增殖抑制作用;细胞转染和Western blot分析细胞自噬;PI染色、Annexin V-FITC/PI和流式细胞术检测细胞周期和细胞凋亡。结果ZST93对K562细胞株的生长呈剂量和时间依赖性的抑制作用,IC_(50)值为2.59μmol·L^(-1),使细胞周期阻滞于G_(1)期。自噬检测中发现ZST93可诱导GFP-LC3积累、LC3-Ⅰ转化为LC3-Ⅱ以及p62表达水平降低。ZST93通过调控自噬激活caspase-8,诱导外部凋亡信号通路,促使caspase-9、caspase-3和PARP剪切而被激活,针对caspase-3的抑制剂Z-DEVD-FMK能够降低ZST93诱导的K562细胞凋亡。结论ZST93可有效抑制K562细胞增殖,促进细胞周期阻滞、细胞凋亡和自噬激活,可能与自噬激活/caspase-8/caspase-3凋亡信号通路有关。
Aim To evaluate the effect of ZST93 on the proliferation in human chronic myeloid leukemia(CML)cells(K562)and explore the possible mechanism.Methods MTT assay,cell growth curve and inverted microscope were used to investigate the effect of ZST93 on proliferation of K562 cells.Cell transfection and Western blot were performed to detect the autophagy,while PI staining,Annexin V-FITC/PI and flow cytometry were conducted to determine cell apoptosis and its anticancer mechanism.Results ZST93 could significantly inhibit the proliferation of K562(IC_(50)=2.59μmol·L^(-1))and induce cell cycle arrest at G_(1)-phase in a dose-and time-dependent manner.Also,through leading to accumulation of GFP-LC3,transition into LC3-Ⅱfrom LC3-Ⅰ,and decrease of p62 expression,ZST93 induced autophagy initiation and autophagic flux.Furthermore,ZST93 induced extrinsic apoptotic pathway by activating caspase-8,and further promoted the cleavage of apoptosis related proteins including caspase-9,caspase-3 and PARP.Moreover,Z-DEVD-FMK,the specific inhibitor of caspase-3,could dramatically reduce the apoptosis induced by ZST93.Taken together,ZST93 could effectively inhibit CML cells,arrest cell cycle at G1-phase,induce cell apoptosis and initiate autophagy.Conclusions The potential mechanism may be related to the regulation of autophagy intiation/caspase-8/caspase-3 signaling pathway,which provides a new idea and theoretical basis for the treatment of CML.
作者
张亚军
黄玖红
何刘军
胡春生
杨东林
陈中祝
唐典勇
ZHANG Ya-jun;HUANG Jiu-hong;HE Liu-jun;HU Chun-sheng;YANG Dong-lin;CHEN Zhong-zhu;TANG Dian-yong(College of Pharmacy,National & Local Joint Engineering Research Center of Targeted and Innovative Therapeutics,Chongqing Key Laboratory of Kinase Modulators as Innovative Medicine, Chongqing University of Arts and Sciences, yongchuan Chongqing 402160, China;College of Pharmaceutical Sciences and Chinese Medicine, Southwest University, Beibei Chongqing 400715, China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2022年第2期215-222,共8页
Chinese Pharmacological Bulletin
基金
重庆市科技局面上项目(No cstc2018jcyjAX0219,cstc2020jcyj-msxmX0733,cstc2020jcyj-msxmX0595)
重庆市教委科学技术研究重点项目(No KJZD-K20200130)。
