大黄素对K562细胞抑制增殖和诱导凋亡的作用  

Effect of Emodin on Proliferation Inhibition and Apoptosis Induction in Leukemic K562 Cells

作　　者：郑志宏[1] 胡建达[1] 陈英玉[1] 连晓岚[1] 郑合勇[1] 郑静[1] 林敏辉[1] 

机构地区：[1]福建医科大学附属协和医院,福建省血液病研究所,福建福州350001

出　　处：《中国实验血液学杂志》2009年第6期1434-1438,共5页Journal of Experimental Hematology

基　　金：福建省科技项目三项费(编号2002Y048);福建省医学创新课题(编号2001-CX-02);福建省高校新世纪优秀人才支持计划(编号NCEFJ-0604);福建省教育厅资助课题(编号JA08095);福建医科大学校级资助课题(编号JS06081)

摘　　要：本研究观察中药大黄素(emodin)对人慢性髓系白血病K562细胞株的增殖及凋亡影响,探讨P210融合蛋白和caspase-3激活在其中的作用。采用MTT法、集落形成试验观察大黄素对K562增殖的影响;Annexin V FITC/PI法、DNA倍体分析及DNA凝胶电泳法检测细胞凋亡;Western blot检测大黄素作用后不同时间段P210、磷酸化P210、caspase-3前体蛋白及PARP表达水平的变化。结果表明,大黄素能抑制K562细胞增殖,作用48小时的半数抑制浓度(IC50)为38.25μmol/L;Annexin V FITC/PI法、亚二倍体峰(凋亡峰)及DNA片段化检测证实,大黄素能诱导K562细胞凋亡,并呈量效关系。大黄素作用K562细胞后P210、磷酸化P210、caspase-3前体蛋白表达水平均有不同程度下调,PARP的活性片段85kD表达增加,这些变化呈时效关系。结论:大黄素能够有效抑制K562细胞增殖并诱导其凋亡。P210磷酸化抑制,P210表达下调和caspase-3激活可能参与了该过程。The study was aimed to investigate the effects of emod/n on proliferation inhibition and apoptosis induction in human chronic myeloid leukemia cell line K562 cells, and to explore the role of P210 protein and activation of caspase 3 in these processes. K562 cells were exposed to emodin at different doses. The proliferation inhibition was detected by MTr assay and colony formation test. The ability of emodin to induce apoptosis and DNA fragmentation were examined by flow eytometry. The expressions of P210, procaspase-3 and PARP protein were determined by Western blot. The results indicated that the emodin remarkably inhibited the K562 cell proliferation, with IC50 value of 38.25 μmol/L after treatment for 48 hours. Meanwhile induced apoptosis, Annexin V-FITC positive cells, sub-G1 apoptotic peak and DNA fragmentation in K562 cells confirmed that emodin induced apoptosis in K562 cells in dose-dependent manner. Western blot results showed that emodin inhibited phosphorylation of P210 protein in K562 ceils and down-regulated the expression levels of P210. The procaspase-3 level in treated K562 cells decreased with increased expressions of PARP in time-dependent manner. It is concluded that the emodin efficiently inhibits growth and induces apoptosis of K562 cells, while the inhibition of phosphorylation of P210 protein, down-regnlation of P210 protein expression and activation of caspase-3 may be involved in these processes.

关 键 词：大黄素 K562细胞 细胞凋亡 P210融合蛋白 CASPASE-3 

分 类 号：R733.72[医药卫生—肿瘤] R979.1[医药卫生—临床医学]