硝酸甘油抑制Hela细胞生长繁殖机制研究

The studies on the inhibitory effect and mechanism of glyceryl trinitrate on Hela cells proliferation

目的 研究硝酸甘油 (GTN)对肿瘤细胞生长繁殖的影响及机制。方法 以人宫颈癌细胞系 (Hela细胞 )为研究对象 ,用四甲基偶氮唑 (MTT)法检测Hela细胞的活性 ;流式细胞仪检测bcl 2蛋白 ,细胞周期 ,细胞凋亡指数 ;比色法检测亚硝酸盐、还原型谷胱甘肽 (GSH)。结果 GTN代谢产生一氧化氮 (NO)及亚硝酸盐 ,MTT试验显示Hela细胞活性明显降低 ;经 0 175mmol/LGTN处理后 ,Hela内GSH由 (43 97± 2 80 )nmol/ 10 6降至 (36 4 6± 2 32 )nmol/ 10 6细胞 ,细胞培养体系GSH由 (71 6 8±3 0 2 ) μmmol/L降至 (5 8 38± 0 81) μmol/L ;bcl 2蛋白表达下降 ,细胞凋亡指数由 1 4 7± 0 0 3提高到2 72± 0 0 89,使Hela细胞G2～M期所占比例由 3 4 7± 1 0 9增加至 12 71± 1 4 5 ,S期所占比例由4 5 95 5± 4 6 6降至 34 86 7± 3 92。结论 GTN可抑制肿瘤细胞的生长繁殖 ,其机制与GTN代谢消耗GSH和产生NO有关。

Objective To investigate the effect of glyceryl trinitrate (GTN) on proliferation of tumor cells and its mechanismMethods Human Cervical Carcinoma Cell Line(Hela) had been used as experimental model MTT assay had been used to evaluate the proliferation activity of Hela cells Bcl-2 protein, cell cycle and apoptosis index in cultured Hela cells had been analyzed by flow cytometer Nitrite and reduced glutathione (GSH) were measured with Griess and DTNB colorimetric methodResults GTN metabolized to nitric oxide (NO) and nitrite and initiated the depressing of proliferation activity in cultured Hela cells When the cultured Hela cells had been treated by 0175 mmol/L GTN,the GSH in cells decreased from (4397±280) nmol/106 to (3646±232) nmol/106 cells and the GSH in cultured medium decreased from 7168±302 to 5838±081 μmol/L,bcl-2 protein decreased and apoptosis index(from 147±003 to 272±0089) increased, cell cycle arrest in G2～M phase had been fond in Hela cells treated with GTNConclusion GTN possess the inhibitory effects on the proliferation of Hela cells and the mechanism was related with the depletion of GSH and releasing of nitric oxide