Zhao Su-Rong, Duan Hai-Feng, Zhang Pei, Liu Hao, Jiang Chen-Chen, Jiang Zhi-Wen
Anhui Engineering Technology Research Center of Biochemical Pharmaceuticals, Anhui Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2013 Apr;21(2):351-5. doi: 10.7534/j.issn.1009-2137.2013.02.019.
This study was purposed to investigate the effects of 2-deoxy-D-glucose (2-DG) on sensitizing HL-60 cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis and its possible mechanism. The proliferative inhibition of HL-60 cells treated with different concentrations of 2-DG and TRAIL was measured by MTT assay. The cells were treated with 2-DG, TRAIL, and 2-DG combined with TRAIL at the concentration < IC50 value, i.e. 10 mmol/L for 2-DG and 100 ng/ml for TRAIL. Apoptosis was analyzed by flow cytometry with PI staining; the expression of RIP1, GRP78, and PARP was analyzed by Western blot; the activity of caspase-3 was detected by special detection kit. The results showed that the combined treatment of HL-60 cells for 48 h induced an apoptotic rate of (45.1 ± 4.3)%, which was significantly higher than that of treated with 2-DG or TRAIL alone; at the same time, the combined treatment potentiated the expression of GRP78 and caspase-3 activity, and down-regulated the expression of RIP1. It is concluded that 2-DG can sensitize HL-60 cells to TRAIL-induced apoptosis, which may be correlated with excessive endoplasmic reticulum stress response, down-regulation of RIP1, and increase of caspase-3 activity.
本研究旨在探讨2-脱氧-D-葡萄糖(2-DG)对HL-60细胞致敏以使其对肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导的凋亡产生影响及其可能机制。采用MTT法检测不同浓度的2-DG和TRAIL处理的HL-60细胞的增殖抑制情况。细胞分别用2-DG、TRAIL以及浓度低于IC50值(即2-DG为10 mmol/L,TRAIL为100 ng/ml)的2-DG与TRAIL联合处理。采用PI染色通过流式细胞术分析凋亡情况;通过蛋白质免疫印迹法分析RIP1、GRP78和PARP的表达;使用专用检测试剂盒检测caspase-3的活性。结果显示,HL-60细胞联合处理48小时诱导的凋亡率为(45.1±4.3)%,显著高于单独使用2-DG或TRAIL处理组;同时,联合处理增强了GRP78的表达和caspase-3的活性,并下调了RIP1的表达。结论是,2-DG可使HL-60细胞对TRAIL诱导的凋亡致敏,这可能与内质网应激反应过度、RIP1下调以及caspase-3活性增加有关。