【摘要】 目的:观察褪黑素对人骨肉瘤细胞系MG63增殖和凋亡的影响及其对细胞周期和Fas基因表达的调控。方法:体外培养MG63细胞,CCK8法检测不同浓度褪黑素对MG63细胞增殖的影响,通过Hoechst 33258染色观察细胞凋亡的形态学变化,流式细胞术检测细胞凋亡及细胞周期改变,实时荧光定量PCR及免疫印迹检测MG63细胞Fas基因表达情况。结果:CCK8法检测显示,褪黑素可抑制MG63细胞增殖,并具有时间和剂量依赖性;Hoechst 33258染色可见凋亡小体;流式细胞术检测显示褪黑素可使凋亡的MG63细胞明显增加,并使细胞周期阻滞在G2/M期;实时荧光定量PCR及免疫印迹分析显示经褪黑素作用后MG63细胞表达Fas蛋白明显增强。结论:褪黑素对MG63细胞的增殖有抑制作用,并诱导其发生凋亡,其机制可能与细胞周期阻滞在G2/M期及上调Fas蛋白表达有关。更多还原

【Abstract】 Objective:To investigate the effect of melatonin on proliferation and apoptosis in the human osteosarcoma cell line MG63 and the regulation on the cell cycle and Fas gene expression.Methods:MG63 cells were cultured in vitro and treated with different concentrations of melatonin.Cell proliferation was detected by CCK8.Hoechst 33258 was used to detect the morphological change of apoptotic cells.MG63 cells were observed by flow cytometry for apoptosis and cell cycle.The expression of Fas was observed by real time fluorescence quantitive PCR and Western blotting.The apoptotic body was observed by Hoechst 33258 dyeing.Results:The proliferation of MG63 cells was inhibited after treatment,which exhibited in a dose and time dependent manner.Compared with the negative control group,the apoptosis rate of MG63 cells was increased and the cell cycle was arrested at the G2/M phase.On the other side,the Fas protein expression was increased significantly.Conclusion:Melatonin can inhibit the proliferation of MG63 cells by inducing its apoptosis.The mechanism may be arresting the cell cycle at the G2/M phase and up-regulating the expression of Fas protein.更多还原