冬凌草甲素抗T细胞急性淋巴细胞白血病效应的实验研究  

Anti-leukemia Effect of Oridonin on T-cell Acute Lymphoblastic Leukemia

作　　者：郭勇[1] 单卿卿[1] 龚玉萍[1] 林娟[1] 杨曦[1] 

机构地区：[1]四川大学华西医院血液科,成都610041

出　　处：《四川大学学报（医学版）》2014年第6期903-907,共5页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(No.30770912);四川省科技厅社会公益项目(No.2008SZ0017);教育部回国留学人员启动金(No.20071108-18-4)资助

摘　　要：目的探讨冬凌草甲素对T细胞急性淋巴细胞白血病的抗白血病效应及其机制。方法以T细胞急性淋巴细胞白血病细胞株CEM为研究对象,应用改良MTT法测定不同浓度及不同时间冬凌草甲素对CEM细胞增殖和生存率的影响,计算72h的半数抑制浓度(IC50);显微镜下观察5、7.5、10μmol/L冬凌草甲素处理24h后CEM细胞的形态学变化;流式细胞术检测0.5、7.5、10μmol/L冬凌草甲素作用24h后CEM细胞的凋亡百分率;应用Western blot法检测7.5μmol/L冬凌草甲素作用后细胞mTOR、P70、4EBP1、RAF、ERK、STAT5信号蛋白水平,以及凋亡调节蛋白Bcl-2和Bax的变化。结果 1冬凌草甲素呈浓度及时间依赖性抑制CEM细胞增殖,作用72h的IC50值为(7.37±1.99)μmol/L;2 5、7.5、10μmol/L冬凌草甲素作用24h后CEM细胞边界不清晰,部分细胞崩解,且浓度越高,细胞形态改变越明显;3 0、5、7.5、10μmol/L冬凌草甲素作用24h后,细胞凋亡百分率分别为(4.8±2.11)%、(19.03±2.54)%、(40.27±3.31)%;(57.23±6.69)%;4冬凌草甲素明显抑制CEM细胞mTOR、P70、4EBP1、RAF、ERK、STAT5信号蛋白的活化;下调CEM细胞抗凋亡蛋白Bcl-2的表达,上调促凋亡蛋白Bax的表达。结论冬凌草甲素可能通过抑制mTOR/P70(4EBP1)、RAF/ERK、STAT5信号蛋白的活化,以及上调促凋亡蛋白Bax、下调抗凋亡蛋白Bcl-2而发挥抗白血病作用。Objective To investigate the antileukemia effect of oridonin on T-cell acute lymphoblastic leukemia cell line CEM.Methods Human T-cell acute lymphoblastic leukemia cell line CEM was cultured in vitro.The 50%inhibition concentration（IC50）of oridonin against CEM cells was examined using modified MTT assay.The cellular morphologic changes were observed using a light microscope.The percent of apoptosis of CEM cells after drug treatment was evaluated by flow cytometric analysis.The active levels of AKT/mTOR,RAF/MEK/ERK,STAT5 signaling pathways and the expression levels of Bcl-2and BAX were examined by Western blot.Results Oridonin inhibited the growth of CEM cells in time-and dose-dependent manner and the IC50 of oridonin was（7.37±1.99）μmol/L after 72 htreatment.The cellular membrane of CEM cells treated with oridonin became unsharp,some of them disintegrated.Oridonin induced apoptosis in CEM cells and the percent of apoptosis rate after 0,5,7.5,10μmol/L oridonin treatment for 24hwere（4.8±2.11）%,（19.03±2.54）%,（40.27±3.31）%and（57.23±6.69）% respectively.Oridonin inhibited activation of mTOR,P70S6,4EBP1,RAF,ERK and STAT5 signaling protein,which were constitutively activated in CEM cells,however,oridonin had no inhibitory effect on AKT kinase.Oridonin down-regulated the level of anti-apoptotic protein Bcl-2and up-regulated the expression of pro-apoptotic protein Bax.Conclusion Oridonin exerted antileukemia effect in CEM cells by inhibiting the activation of mTOR/P70/4EBP1,RAF/ERK and STAT5 signaling pathways,down-regulating the expression of Bcl-2and up-regulating the expression of BAX.

关 键 词：CEM细胞株 抗白血病效应 冬凌草甲素 AKT/MTOR RAF/MEK/ERK STAT5 

分 类 号：R733.71[医药卫生—肿瘤]