摘要
目的:研究SU11248(舒尼替尼)诱导慢性骨髓性白血病K562细胞凋亡的分子机制。方法:采用CCK-8比色检测SU11248对K562细胞增殖的影响;流式细胞术(FCM)检测K562细胞周期变化;间接免疫荧光检测凋亡相关蛋白的表达及定位;免疫印迹检测凋亡相关蛋白的表达变化。结果:CCK-8比色显示SU11248可明显抑制K562细胞增殖(P<0.05),呈现剂量和时间依赖性。FCM显示该药可阻滞K562细胞于G0/G1期。间接免疫荧光染色可见SU11248组细胞色素C(Cyto C)在胞质中呈弥散或粗块状分布,而p53、p73及NF-κB p65均主要定位于胞质,较对照组表达差异不明显。免疫印迹显示,随时间延长,SU11248组较对照组Bcl-2表达呈下降趋势,Bax和Cyto C表达呈增加趋势,同时有Caspase-9和Caspase-3的激活,而p53、p73及NF-κB p65表达变化不明显。结论:SU11248可通过阻滞细胞周期进程及激活内源性线粒体凋亡通路诱导K562细胞调亡。
Objective：To explore the molecular mechanisms of SU11248 （Sunitinib） induced apoptosis on chronic myeloid leukemia cell line K562 cell line. Methods：The effects of SU11248 on K562 cell-growth were estimated by cell counting kit 8 （CCK-8）. Flow cytometry was employed to detect the cell cycle changes. Indirect immunofluorescence was used to observe the expression and location of apoptosis related proteins in K562 cells. The expression of apoptosis related proteins were identified hy Western blot. Results： CCK-8 tests showed SU11248 could inhibit the growth of K562 cells remarkably （P 〈 0.05 ） in a time- and dose-dependent manner. Flow cytometry test indicated this drug could block the cell cycle in G0/G1 phase. Indirect immunofluorescence unfolded that cytochrome C（ Cyto C） of SU11248 group located in cytoplasm with a dispersive or block like distribution, while p73, p53 and NF-KB p65 located in cytoplasm equably with no difference. Western blot revealed that compared with the control group the expression level of Bcl-2 protein in SUl1248 group decreased, while, Bax and Cyto C increased as well as the activation of Caspase-9 and Caspase-3 with the time prolonged. But, p53, p73 and NF-KB p65 changed unconspicuously. Conelusion：SUl1248 could induce apoptosis of K562 cell remarkably, and the mechanisms could be linked to its functional role in blocking the cell cycles and activating the endogenous mitochondria pathways.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2012年第7期584-588,共5页
Chinese Journal of Immunology
基金
福建省教育厅资助项目(JA07085)
福建医科大学教授基金资助项目(JS08009)
