2-脱氧葡萄糖可增强TRAIL诱导的口腔癌细胞凋亡的敏感性

Glycosylation inhibitor 2-deoxy-D-glucose sensitizes oral cancer cells to TRAIL-induced apoptosis

目的探讨糖基化抑制剂2-脱氧葡萄糖(2-DG)对肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导肿瘤细胞凋亡作用的影响。方法 MTT法检测不同浓度(0、0.625、1.25、2.5、5、10 mmol/L)2-DG及不同浓度2-DG与TRAIL(200 ng/ml)合用对口腔癌细胞KB的增殖抑制作用。集落克隆法检测2-DG及TRAIL对口腔癌细胞KB的增殖抑制作用。溴化丙啶(PI)单染法检测2-DG(5 mmol/L)对TRAIL诱导口腔癌细胞KB凋亡的影响;Western blot检测2-DG(5 mmol/L)处理口腔癌细胞KB不同时间(0、6、16、24 h),DR5的表达以及联合TRAIL处理后Caspase-3的表达。结果 5 mmol/L 2-DG作用于口腔癌细胞KB 24、48、72 h细胞存活率分别为75.25%、69.06%、59.19%,但24 h细胞凋亡率仅为15.9%。5 mmol/L 2-DG与TRAIL联合作用于口腔癌细胞KB 24 h的凋亡率为72.5%,高于TRAIL本身诱导凋亡率45.3%,并且2-DG可增强TRAIL抑制口腔癌细胞KB的集落克隆形成的作用。2-DG上调DR5的表达并且增强Caspase-3的激活。结论 2-DG能增强TRAIL诱导口腔癌细胞的凋亡,其机制可能是上调DR5的表达及增强Caspase-3的激活。

Objective To investigate the effect of 2-deoxy-D-glucose（2-DG） in enhancing the sensitivity of oral cancer cells to tumor necrosis factor-related apoptosis-inducing ligand（TRAIL）-induced apoptosis.Methods The oral cancer cell line KB was incubated in the presence of different concentrations（0,0.625,1.25,2.5,5,and 10 mmol/L） of 2-DG with or without TRAIL（200 ng/ml）.The cell viability was measured using MTT assay and cell apoptosis was detected using flow cytometry with propidium iodide（PI） staining.KB cells treated with 5 mmol/L 2-DG with or without TRAIL for 0,6,16,or 24 h were examined with Western blotting for protein expressions of death receptor 5（DR5） and caspase-3.Results Treatment of the cells with 5 mmol/L 2-DG for 24,48 and 72 h resulted in a cell viability of 25.25%,69.06%,and 59.19%,respectively.Combined treatment with 5 mmol/L 2-DG with TRAIL for 24 significantly enhanced the cell apoptotic rate（72.5%） as compared to the rate induced by TRAIL alone（45.3%） and by 2-DG（15.9%） alone.2-DG treatment markedly up-regulated DR5 and caspase-3 expression and enhanced the inhibitory effect of TRAIL on cell colony formation.Conclusion 2-DG sensitizes oral cancer cells to TRAILinduced apoptosis by up-regulating DR5 and caspase-3 expressions.