siRNA抑制缺氧诱导因子-1ɑ表达对人视网膜母细胞瘤细胞增殖与凋亡的影响

siRNA抑制缺氧诱导因子-1ɑ表达对人视网膜母细胞瘤细胞
增殖与凋亡的影响
郎莉莉;高玉;葛茸茸;陈建梅;景明
【期刊名称】《海军医学杂志》
【年(卷),期】2016(037)001
【摘要】目的观察siRNA抑制人视网膜母细胞瘤表达低氧诱导因子( HIF)对细胞增殖和凋亡的影响. 方法以Y-79细胞系为研究对象,分别在正常氧和低氧环境下培养,MTT法检测细胞增殖,RT-PCR和Western blot 检测HIF-1ɑ mRNA和蛋白质水平的表达变化,siRNA抑制细胞表达HIF-1ɑ,膜联蛋白V-FITC和PI双染法流式
细胞术检测细胞凋亡情况,荧光评估测定法检测半胱天冬酶活性. SPSS 16.0软件包作统计学分析,配对t检验. 结果与常氧和低氧环境培养Y79细胞比较,在0、4、8、12 h后HIF-1ɑmRNA和蛋白质数倍高表达. siRNA干扰技术处理24、48、72 h 后,常氧环境培养下的Y-79细胞表达HIF-1ɑ呈时间依赖性轻度下降趋势,低氧条件下呈显著下降. siRNA HIF-1ɑ可能是通过上调Bax/Bcl-2 比率和活化caspase-9、caspase-3途径促进Y-79细胞凋亡. 结论 siRNA HIF-1ɑ下调人视网膜母细胞瘤
表达低氧诱导因子,可抑制细胞增殖和促进细胞凋亡. 抑制HIF-1ɑ表达可能是治疗视网膜母细胞瘤的潜在新策略.%Objective To investigate the effect of hypoxia inducible factor-1ɑknocked down by small interfering RNA ( siR-NA) on
cell proliferation , apoptosis and apoptotic pathways of human Y-79 RB cells.Methods Cell proliferation , HIF-1ɑmRNA and protein levels were measured by MTT , RT-PCR and Western Blot on human Y-79 RB cells under both normoxic and hypoxic conditions , respectively .siRNA
knockdown against HIF-1ɑwas carried out to suppress the expression of HIF-1ɑ.Cell apotosis was determined by double staining cells with the annexin V-FITC and PI.Caspase activity was assessed by the fluorometric assay .Biostatistical analyses were conducted with SPSS 16.0 software package.Results A multifold increase in HIF-1ɑmRNA and protein levels were observed af-ter cells were exposed to the hypoxic environment at 0 h,4 h,8 h and 12 h.Both mRNA and protein levels of HIF-1ɑwere attenuated or abolished by transfection with siRNA H IF-ɑunder normoxic and hypoxic conditions .Futhermore,knockdown of HIF-1ɑcould enhance hy-poxia-induced slight increase of Bax/Bcl-2 ratio and activate caspase-9 and caspase-3.Conclusion This study was able to show that a knockdown of HIF-1ɑby siRNAH IF-1ɑresulted in a decrease in proliferation and induction of apoptosis in human Y-79 RB cells under both normoxic and hypoxic conditions .HIF-1ɑexpression may be a promising strategy for the treatment of human RB in the future .
【总页数】5页(P22-26)
【作者】郎莉莉;高玉;葛茸茸;陈建梅;景明
【作者单位】200120 上海,同济大学附属东方医院眼科;200120 上海,解放军第四一一医院眼科;200120 上海,解放军第四一一医院眼科;200120 上海,同济大学附属东方医院眼科;200120 上海,解放军第四一一医院眼科
【正文语种】中文
【中图分类】R774
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