氟诱导成骨细胞MC3T3-E1自噬与凋亡及其相互作用分析

目的主要研究氟诱导成骨细胞MC3T3-E1发生自噬、凋亡及两者之间的关系。方法利用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)比色法检测氟对成骨细胞MC3T3-E1的增殖活性影响,探寻氟引起凋亡的浓度;流式细胞术检测细胞凋亡;western blot免疫印记技术检测凋亡相蛋白;利用western blot免疫印记技术检测Beclin 1蛋白及LC3蛋白表达水平及变化。结果氟能明显抑制成骨细胞MC3T3-E1增殖,诱发细胞凋亡,其抑制作用呈时间、剂量依赖性;氟在促进细胞凋亡的同时也促进自噬产生;3-甲基腺素(3-MA)抑制自噬后,氟诱导的凋亡进一步增多。结论氟明显抑制MC3T3-E1细胞增殖,同时诱发细胞凋亡和自噬;氟诱导的自噬部分拮抗其诱导的凋亡。

An analysis on fluorine-induced autophagy and apoptosis of MC3T3-E1 and their interaction

Objectives: To explore Fluorine-induced apoptosis and autophagy of MC3T3-E1 cells and the relationship between them.Methods To explore the concentration of fluorine that caused cell apoptosis the influence of fluorine on proliferative vitality of MC3T3-E1 was assessed by MTT assay.The apoptosis of MC3T3-E1 was detected by flow cytometry;The expression of apoptosic protein was determined by Western Blot;The LC3 and the Beclin1 protein level and variation were analyzed by western blot assay.Results Fluorine inhibited the proliferation of MC3T3-E1 cells dose and time dependently and simultaneously induced apoptosis and autophagy;Inhibition of autophagy by 3-MA promotes fluorine-induced apoptosis.Conclusion Fluorine can inhibit cell proliferation and induce apoptosis and autophagy,but autophagy partly atagonizes apoptosis.