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双氢青蒿素对人骨肉瘤细胞株143B增殖和凋亡的作用
引用本文:刘跃亮,罗进勇,张彦,何通川,曾照芳.双氢青蒿素对人骨肉瘤细胞株143B增殖和凋亡的作用[J].中国药理学通报,2012,28(12):1719-1723.
作者姓名:刘跃亮  罗进勇  张彦  何通川  曾照芳
作者单位:1. 重庆医科大学检验医学院,临床检验诊断学教育部重点实验室,重庆,400016
2. 美国芝加哥大学医学中心分子肿瘤学研究室,芝加哥,60637
基金项目:国家自然科学基金资助项目
摘    要:目的观察双氢青蒿素(dihydroarteminin,DHA)对人骨肉瘤细胞143B的增殖和凋亡的影响以及可能的机制。方法体外培养人骨肉瘤细胞株143B;MTT比色法和克隆形成实验检测不同浓度DHA对骨肉瘤细胞存活与克隆形成能力的影响。Hoechst 33258染色法观察细胞凋亡的形态变化。构建β-Catenin荧光素酶报告基因(β-Catenin-Luc,p-Top)检测DHA作用143B后细胞内β-Catenin活性变化。不同浓度的DHA作用后,Western blot检测与细胞增殖(如PC-NA、Cyclin D1、c-Myc)、凋亡(如Bad、Bcl-2、Caspase-3)密切相关的标志物蛋白质表达变化。结果 DHA作用于143B细胞24 h后,MTT结果显示143B细胞的增殖活性受到明显抑制,且其克隆形成能力减弱(P<0.05),在DHA浓度达35μmol.L-1时,抑制效率最明显。Western blot结果显示PC-NA表达下调;而促凋亡蛋白Bad和Caspase-3表达上调,Bcl-2蛋白表达明显减弱。结论双氢青蒿素具有较明显的抑制人骨肉瘤细胞的增殖且促进其凋亡的作用,可能通过下调细胞增殖相关蛋白PCNA、Bcl-2和上调促凋亡蛋白Bad和Caspase-3,启动凋亡程序,致143B细胞发生凋亡。

关 键 词:双氢青蒿素  人骨肉瘤细胞  细胞增殖  细胞凋亡  Wnt信号通路  β-Catenin

Role of dihydroartemisinin(DHA) in proliferation and apoptosis of human osteosarcoma 143B cells
LIU Yue-liang , LUO Jin-yong , ZHANG Yan , HE Tong-chuan , ZENG Zhao-fang.Role of dihydroartemisinin(DHA) in proliferation and apoptosis of human osteosarcoma 143B cells[J].Chinese Pharmacological Bulletin,2012,28(12):1719-1723.
Authors:LIU Yue-liang  LUO Jin-yong  ZHANG Yan  HE Tong-chuan  ZENG Zhao-fang
Institution:1(1.College of Laboratory Medicine Chongqing Medical University,Key Laboratory of Clinical Laboratory Diagnostics of Ministry of Education,Chongqing 400016,China;2.Cancer Research Center the University of Chicago,60637 USA)
Abstract:Aim To observe the effect of dihydroartemisinin(DHA) on proliferation and apoptosis of human osteosarcoma 143B cells and the possible mechanism.Methods Human osteosarcoma cell line 143B was cultured in vitro;MTT 3-(4,5) the-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide,MTT] colorimetric assay and clone formation assay were applied to detect the role of DHA in the survival and growth status of human osteosarcoma cells.Hoechst 33258 cell staining was used for apoptosis detection.The luciferase reporter gene,β-Catenin-Luc(p-Top) was constructed to detect β-Catenin activity after DHA's role in 143B cells.The marker protein expression closely related to tumor cell proliferation,such as PCNA,Cyclin D1,C-Myc,and apoptosis,such as Bad,Bcl-2,Caspase-3,was detected by Western blot.Results 24 hours after DHA played role in 142B,MTT results showed that DHA could inhibit the proliferation of the 143B cells,and colony formation diminished(P<0.05),the inhibition efficiency increased significantly at a concentration of 35 μmol·L-1DHA.143B cells was significantly enhanced(P<0.05),and the PCNA expression was lowered,and cell proliferation was inhibited,pro-apoptotic protein Bad and caspase-3 expression was significantly enhanced,while Bcl-2 protein expression decreased significantly.Conclusions Dihydroartemisinin significantly inhibits the proliferation of human osteosarcoma 143B cells and promote apoptosis,which may be associated with the downregulation of cell proliferation-associated proteins,the PCNA and Bcl-2,and the upregulation of pro-apoptotic protein,Bad and Caspase-3.
Keywords:dihydroarteminin(DHA)  human-osteosarcoma  proliferation  apoptosis  Wnt signaling pathway  β-Catenin
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