| 摘要: |
| 【目的】运用siRNA沉默小鼠前体脂肪细胞3T3-L1的细胞信号负调控因子3基因(SOCS3)，探讨SOCS3在肿瘤坏死因子-α(TNF-α)诱导3T3-L1细胞凋亡中的作用。【方法】体外培养3T3-L1细胞，用0,20,40,60,80,100,150,200 ng/mL TNF-α处理细胞24 h后,观察细胞凋亡情况。利用脂质体LipofectamineTM 2000，用化学合成的SOCS3小干扰 RNA(siRNA)转染3T3-L1细胞,用100 ng/mL TNF-α刺激24 h,荧光显微镜下观察细胞凋亡的变化,RT-PCR检测SOCS3、c-myc和survivin mRNA的表达情况。【结果】SOCS3 siRNA抑制了3T3-L1细胞中SOCS3 mRNA的表达(P＜0.05)。与空白对照组相比,SOCS3 siRNA组的SOCS3 mRNA表达量极显著减少,c-myc和survivin mRNA的表达水平极显著或显著升高,而脂质体组和阴性对照组无显著差异。【结论】体外试验结果证明, 靶向抑制SOCS3基因表达可以有效抑制TNF-α诱导的3T3-L1细胞凋亡。 |
| 关键词: RNA干扰 细胞信号负调控因子3基因 TNF-α 细胞凋亡 3T3-L1前体脂肪细胞 |
| DOI： |
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| 基金项目:国家自然科学基金项目(30871785) |
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| Silencing SOCS3 expression’s inhibitition on TNF-α induced apoptosis in 3T3-L1 preadipocytes |
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| Abstract: |
| 【Objective】The experiment investigated the influence of SOCS3 siRNA on TNF-α induced apoptosis in 3T3-L1 preadipocytes.【Method】3T3-L1 cells were cultured in vitro and treated with TNF-α at the concentrations of 0,20,40,60,80,100,150,200 ng/mL for 24 h,respectively.Optical microscope was used to observe morphological changes during apoptosis.Chemically synthesized small interfering RNA (siRNA) directed against SOCS3 gene was transfected into 3T3-L1 cells by cationic liposome LipofectamineTM 2000,then the cells were treated with TNF-α at 100 ng/mL for 24 h.The cells apoptosis was evaluated by fluorescence microscope,and the expression of SOCS3,c-myc and survivin mRNA was investigated by RT-PCR.【Result】SOCS3 gene was knocked down effectively by SOCS3 siRNA in 3T3-L1 cells(P<0.05).Compared with control group,in SOCS3 siRNA group the expression of SOCS3 mRNA decreased extremely significantly,the expression of c-myc mRNA increased extremely significantly,and the expression of survivin mRNA increased remarkably,while no distinct change was detected in liposome group or negative control group.【Conclusion】These results suggest that knocking down SOCS3 gene by RNAi can effectively inhibit TNF-α induced apoptosis in 3T3-L1 cells. |
| Key words: RNA interference SOCS3 gene tumor necrosis factor α apoptosis 3T3-L1 preadipocytes |
