XIAP抑制剂增强依托泊苷对急性淋巴细胞白血病6T⁃CEM细胞凋亡诱导作用的研究

XIAP inhibitor enhances apoptosis induction of etoposide on acute lymphoblastic leukemia 6T⁃CEM cells

目的研究XIAP抑制剂AT-406联合依托泊苷对急性淋巴细胞白血病6T-CEM细胞凋亡的诱导作用及分子机制.方法体外培养6T-CEM细胞,加入不同浓度(0.1、1、10μmol/L)的XIAP抑制剂AT-406和20 nM依托泊苷处理6T-CEM细胞,CCK8分析药物对细胞生长抑制率;PE-Annexin-V/7-AAD双染检测XIAP抑制剂AT-406联合依托泊苷对6T-CEM细胞凋亡的影响;比色法分析XIAP抑制剂AT-406联合依托泊苷处理的6T-CEM细胞中Caspase-3酶活性的影响,Western Blot分析凋亡相关蛋白Cleaved-Caspase-3表达水平.结果不同浓度XIAP抑制剂AT-406处理6T-CEM细胞后明显抑制细胞的增殖,AT-406增强依托泊苷对细胞的抑制作用;流式细胞术检测细胞凋亡,XIAP抑制剂AT-406增强可以依托泊苷对6T-CEM细胞凋亡的作用诱导,差异具有统计学意义(P<0.05);AT-406联合依托泊苷增强6T-CEM细胞中Caspase-3酶活性;Western Blot实验结果显示XIAP抑制剂AT-406处理的6T-CEM细胞中,细胞凋亡执行蛋白Cleaved-Caspase-3表达明显升高,差异具有统计学意义(P<0.05).结论XIAP抑制剂AT-406增强依托泊苷对急性淋巴白血病6T-CEM细胞凋亡的诱导作用,激活细胞凋亡途径.

Objective To study the effect and molecular mechanism of XIAP inhibitor AT⁃406 combined with etoposide on apoptosis of 6T⁃CEM cells in acute lymphoblastic leukemia.Methods 6T⁃CEM cells were cultured in vitro and treated with different concentrations of XIAP inhibitors AT⁃406 and 20nM etoposide,CCK8 was used to analyze the growth inhibitory rate of 6T⁃CEM cells.PE⁃Annexin⁃V/7⁃AAD double staining was used to detect the effect of XIAP inhibitor AT⁃406 combined with etoposide on the apoptosis of 6T⁃CEM cells.The enzymatic activity of Caspase⁃3 in 6T⁃CEM cells treated with XIAP inhibitor AT⁃406 combined with etoposide was analyzed by colorimetric assay,and the effects on the expression of apoptosis⁃related protein Cleaved⁃Caspase⁃3 was analyzed by Western Blot.Results Different concentrations of XIAP inhibitor AT⁃406 significantly inhibited the proliferation of 6T⁃CEM cells,AT⁃406 enhanced the inhibitory effect of etoposide on cells;Flow cytometry was used to detect apoptosis,and enhanced XIAP inhibitor AT⁃406 could induce the apoptosis of 6T⁃CEM cells,the difference was statistically significant(P<0.05).Combination of etoposide enhanced the activity of Caspase⁃3 in 6T⁃CEM cells.Western blot results showed that the expression of Cleaved⁃Caspase⁃3 in 6T⁃CEM cells treated with XIAP inhibitor AT⁃406 increased significantly(P<0.05).Conclusions XIAP inhibitor AT⁃406 enhances the apoptotic induction of etoposide in 6T⁃CEM cells of acute lymphoblastic leukemia and activates the apoptotic pathway.