摘要
目的:探讨大黄素对人白血病K562细胞的增殖抑制与诱导凋亡作用。方法:采用MTT法检测大黄素对K562细胞的增殖抑制作用;通过Wight-Giemsa染色(W-G染色),在普通光镜下观察细胞的形态学变化;运用AnnexinV/PI双标记法检测大黄素对K562细胞的凋亡效应;采用流式细胞技术检测细胞周期变化与凋亡情况;通过比色法检测Caspase-3、8、9的相对活性,分析大黄素诱导K562细胞凋亡的信号途径。结果:大黄素能显著抑制K562细胞的增殖,作用24、48、72h后的半数抑制率浓度分别为80、50、40μmol/L;处理组细胞在普通光镜下可见典型的凋亡形态学改变;AnnexinV/PI双标记法检测到K562细胞在大黄素的诱导下出现早期凋亡并呈剂量依赖性(P<0.01);流式细胞仪结果分析发现,处理组的K562细胞被阻滞于G0/G1期,与对照组相比,凋亡率明显升高并呈现剂量依赖性(P<0.01);大黄素可触发Caspase-3、8、9的活性增高(P<0.01)。结论:大黄素能有效地抑制K562细胞的增殖并诱导其凋亡,其机制和Caspase信号途径的活化有关。
Objective: To study the mechanism of K562 leukemia ceils proliferation and apoptosis induced by emodin. Methods: Inhibitory effect of emodin on proliferation of K562 leukemia cells was assayed by MTT method. The morphologie changes of K562 cells were observed under microscop after Wight-Giemsa staining. Apoptosis was checked by Annexin V/PI. The changes of cell cycle and apoptosis were detected by flow cytometry. The activities of Caspase-3,8,9 were checked by chromatometry to analyze the apoptosis of K562 cells. Results: The proliferation of K562 leukemia cells was significantly inhibited by emodin. The IC50 of K562 cells inhibited with emodin for 24 h, 48 h and 72 h were 80,50 and 40 μ mol/L respectively. Typical morphological changes of K562 cells were observed by microscopy. The proliferation of K562 cells was obviously inhibited in dose-dependent manner. In Annexin V/PI, emodin induced K562 cells toward apoptosis in dose-dependent manner (P〈0.01). The result of flow cytometry indicated that the cell cycle was blocked in G0/G1 phase (P〈0.01). After treated with emodin, the activities of Caspase-3,8,9 were significantly higher than those in the normal control (P〈0.01 ). Conclusion: Emodin inhibits proliferation and induces apoptosis of K562 leukemia cells, and up-regulating the experession of Caspase may be one of its mechanisms.
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2009年第7期822-825,共4页
Journal of Chongqing Medical University
基金
国家自然科学基金资助项目(30300449)
国家中医药管理局资助项目(02-03ZP52)
重庆医科大学校级课题资助项目(XBYB200710
XBYB2007108)