Rigosertib对白血病细胞系HEL和K562的抗肿瘤作用及机制研究  

Anti-Tumor Effect of Rigosertib on HEL and K562 Cells and Its Related Mechanism

作　　者：许峰[1] 常春康[1] 李晓[1] 贺琪[1] 吴凌云[1] 张征[1] 

机构地区：[1]上海交通大学附属第六人民医院血液科,上海200233

出　　处：《中国实验血液学杂志》2017年第5期1362-1366,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目(81270584;81300389;81470291)

摘　　要：目的:探讨Rigosertib对白血病细胞系HEL和K562细胞的凋亡、增殖和细胞周期的影响及其作用机制。方法:Rigosertib梯度浓度作用于HEL和K562细胞,采用Annexin V/PI双染流式细胞术检测细胞凋亡,WST-1法绘制细胞增殖曲线,PI染色流式细胞术检测细胞周期,流式细胞术胞内染色检测胞内信号通路蛋白表达。结果:Rigosertib明显诱导HEL和K562细胞凋亡且呈现时间依赖性(r=0.997)和剂量依赖性(r=0.987);以低剂量Rigosertib浓度(0.5μmol/L)分别作用于HEL和K562细胞6-54 h,明显抑制HEL和K562细胞增殖,诱导HEL和K562细胞阻滞于 G_0/M期, G_1期细胞比例明显降低。流式细胞术分析显示,Rigosertib激活胞内凋亡蛋白cleaved caspase 3和cleaved PARP蛋白的表达,同时抑制增殖蛋白BCL-2和Cyclin D1蛋白的表达。此外Rigosertib明显抑制AKT、磷酸化AKT(S473)和磷酸化GSK-3α/β(S21/9)的表达。结论:Rigosertib诱导白血病细胞系HEL和K562细胞凋亡,增殖抑制和细胞周期阻滞,其抗肿瘤作用可能通过抑制AKT-GSK信号通路而实现的。本研究为rigosertib进一步应用于临床前研究提供了实验依据。Objective： To investigate the effects of rigosertib on the apoptosis,proliferation and cell cycle of HEL and K562 cells. Methods： The HEL and K562 cells were treated with different concentration of rigosertib at different time points,the cell apoptosis,proliferation and cycle were determined by using flow cytometry with Annexin V/PI double staining,WST-1 method and 7-AAD assay,respectively. Intracellular signaling proteins were detected by flow cytometry（FCM）. Results： Rigosertib induced obvious apoptosis in HEL and K562 cells,and the apoptotic effect was both time-dependent and dose-dependent manner（ P〈0.05）. The low dose of rigosertib inhibited obviously the proliferation of HEL and K562 cells after treatment from 6 to 54 h,Rigosertib arrested HEL and K562 cells into G2/M phase. In addition,Rigosertib obviously increased the expression of apoptosis-related proteins such as cleaved caspase 3 and PARP,and reduced the proliferation-related proteins such as BCL-2 and Cyclin D1. Rigosetib inhibited the activation of AKT-GSK signaling through decreasing the expression of AKT,pAKT（Ser473） and GSK-3α/β（S21/9）.Conclusion： Rigosertib inhibites proliferation,induces apoptosis and cell cycle arrest in G2/M phase of HEL and K562 cells. This agent may have potential application prospect in leukemia therapy.

关 键 词：rigosertib HEL细胞系 K562细胞系 细胞增殖 细胞凋亡 细胞周期 AKT-GSK信号通路 

分 类 号：R733.7[医药卫生—肿瘤] R979.1[医药卫生—临床医学]