Notch1 siRNA对乏氧环境骨肉瘤细胞MG-63增殖及周期的影响Effects of Notch 1 siRNA on the cell cycle and proliferation of the osteosarcoma cell line MG-63 under hypoxic conditions
李纯璞;陈元振;尚凯;郭冬梅;张开刚;
摘要(Abstract):
[目的]探讨Notch1 siRNA对乏氧环境人骨肉瘤细胞MG-63细胞增殖和周期的影响。[方法]将骨肉瘤细胞株MG-63细胞置于乏氧(37℃、1%O2、5%CO2)环境下培养作为实验组,以正常氧环境(37℃、21%O2、5%CO2)下培养作为对照组,培养72 h,应用MTT方法检测不同时间乏氧培养对MG-63细胞增殖的影响;采用Notch siRNA转染乏氧环境中的MG-63细胞,MTT方法检测细胞增殖变化;流式细胞术检测细胞周期的变化;Western blot检测细胞HIF-1a、Notch1蛋白表达变化。[结果]乏氧培养可促进MG-63细胞增殖,48 h常氧及乏氧对细胞增殖的作用可见明显差异(2.164±0.075 vs 1.421±0.086)(P<0.01)。流式细胞分析显示实验组、对照组MG-63细胞培养48 h G1/G2期细胞比例分别为(41.79±3.25)%和(53.87±2.31)%(P<0.05),乏氧可以促进细胞周期进展,Notch1 siRNA有效下调乏氧MG-63细胞Notch1蛋白表达,Notchl siRNA作用48 h后乏氧环境MG-63细胞周期阻滞于G1/G2期。Western-blot结果显示乏氧环境下MG-63细胞HIF-1a、Notch1蛋白表达增高。[结论]乏氧环境能通过促进MG-63细胞周期进展,从而明显促进细胞增殖,阻断Notch可逆转乏氧对MG-63细胞促增殖作用,提示Notchl是治疗骨肉瘤的有效分子靶点。
关键词(KeyWords): 骨肉瘤;乏氧;细胞增殖;细胞周期;Notch信号通路
基金项目(Foundation): 山东省自然科学基金(编号:ZR2012HL38、2009ZRA09006);; 山东省优秀中青年科学家科研奖励基金(编号:BS2013SF030);; 山东省医药卫生科技发展计划项目(编号:2011HW077)资助
作者(Author): 李纯璞;陈元振;尚凯;郭冬梅;张开刚;
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