Xu Feng, Chang Chun-Kang, Li Xiao, He Qi, Wu Ling-Yun, Zhang Zheng
Department of Hematology, Sixth People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai 200233, China.
Department of Hematology, Sixth People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai 200233, China. E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2017 Oct;25(5):1362-1366. doi: 10.7534/j.issn.1009-2137.2017.05.014.
To investigate the effects of rigosertib on the apoptosis, proliferation and cell cycle of HEL and K562 cells.
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).
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 G/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).
Rigosertib inhibites proliferation, induces apoptosis and cell cycle arrest in G/M phase of HEL and K562 cells. This agent may have potential application prospect in leukemia therapy.
研究瑞戈非尼对HEL细胞和K562细胞凋亡、增殖及细胞周期的影响。
在不同时间点用不同浓度的瑞戈非尼处理HEL细胞和K562细胞，分别采用Annexin V/PI双染流式细胞术、WST-1法和7-AAD检测法测定细胞凋亡、增殖及细胞周期。通过流式细胞术（FCM）检测细胞内信号蛋白。
瑞戈非尼诱导HEL细胞和K562细胞明显凋亡，且凋亡效应呈时间和剂量依赖性（P<0.05）。低剂量瑞戈非尼在处理6至54小时后明显抑制HEL细胞和K562细胞的增殖，瑞戈非尼使HEL细胞和K562细胞停滞于G/M期。此外，瑞戈非尼明显增加凋亡相关蛋白如裂解的半胱天冬酶3和PARP的表达，并降低增殖相关蛋白如BCL-2和细胞周期蛋白D1的表达。瑞戈非尼通过降低AKT、pAKT(Ser473)和GSK-3α/β (S21/9)的表达抑制AKT-GSK信号通路的激活。
瑞戈非尼抑制HEL细胞和K562细胞的增殖，诱导凋亡并使细胞周期停滞于G/M期。该药物在白血病治疗中可能具有潜在的应用前景。
