SOCS 3基因对缺氧肺动脉平滑肌细胞增殖及原癌基因mRNA表达的影响  

Effects of cytokine signaling 3 suppressors gene on c-fos and c-jun mRNA expression and proliferation of pulmonary arterial smooth muscle cells in rat under hypoxia

作　　者：白莉[1] 余祖滨[1] 钱频[1] 钱桂生[1] 关崧[1] 李淑萍[1] 

机构地区：[1]第三军医大学新桥医院全军呼吸内科研究所,重庆400037

出　　处：《中华内科杂志》2005年第1期42-45,共4页Chinese Journal of Internal Medicine

基　　金：国家自然科学基金资助项目 (3 9970 3 2 9)

摘　　要：目的　探讨细胞因子信号负调控因子 (SOCS) 3基因对缺氧大鼠肺动脉平滑肌细胞(PASMCs)原癌基因c fos、c junmRNA表达及细胞增殖的影响。 方法　实验分 pEFSOCS3和pSV2neo共转染组 (A组 )、未转染组 (B组 )、pEF和 pSV2neo共转染组 (C组 )。以脂质体为载体将pEFSOCS3和pSV2neo共转染至体外培养的PASMCs中 ,用免疫细胞化学法测转染前后细胞中SOCS3蛋白表达 ;用半定量RT PCR检测转染前后常氧、缺氧 2、4、6、8、12h细胞c fos、c junmRNA表达水平 ;流式细胞仪测转染前后上述各时相点细胞周期的变化。结果　免疫细胞化学法证实转染后细胞中有SOCS3稳定表达。半定量RT PCR结果 ,B、C组缺氧 2hc fosmRNA表达水平达高峰 ,4h下降 ,6h降至正常 ,8h再次达高峰 ,12h下降 ;与B组同时相点相比 ,A组缺氧 2、8hc fosmRNA表达水平低 (P <0 0 1)。B、C组缺氧 2hc junmRNA表达水平升高 ,6h达高峰 ;与B组同时相点相比 ,A组缺氧 2、4、6、8hc jun表达水平低 (P <0 0 1)。B、C组缺氧 6h即出现G0 /G1期细胞比例减少 ,S +G2 /M期细胞比例增加 (P <0 0 1) ;与B组同时相点相比 ,A组G0 /G1期细胞比例增多 ,S +G2 /M期细胞比例减少 (P <0 0 1)。结论　缺氧可能通过诱导PASMCsc fos、c jun表达而促进PASMCs增殖 ;SOCS3蛋白可能通?Objective To explore the effects of suppressors of cytokine signaling (SOCS)3 gene on expression of c fos, c jun mRNA and proliferation of rat pulmonary arterial smooth muscle cells(PASMCs) under hypoxia.Methods PASMCs were co transfected with pEFSOCS3 and pSV2neo by liposome, and then expression of SOCS3 protein was detected by immunocytochemistry. After PASMCs were exposed to normoxic and hypoxia at various time points respectively,expression of c fos and c jun mRNA was assessed by semi quantitive RT PCR. Flow cytometric DNA analysis was used to detect cell cycles.Results Expression of SOCS3 protein was confirmed by Western blot in PASMCs transfected with SOCS3 gene. The c fos mRNA level in control cells peaked at 2 h of hypoxia and declined at 4 h, then peaked at 8 h secondly and declined at 12 h. C fos mRNA level in SOCS3 gene transfected cells at 2 h and 8 h exposed to hypoxia was lower than that in control cells at the same time points respectively ( P <0 01). The c jun mRNA level increased at 2 h after exposure of control cells to hypoxia, peaked at 6 h of hypoxia and declined to the basal levels at 12 h. C jun mRNA level of SOCS3 gene transfected cells at 2 h,4 h,6 h,8 h under hypoxia was lower than that in controls cells at the same time points. Compared with control PASMCs, cells in transfected with SOCS3 gene at G 1/G 0 phase increased and those at S+G 2/M phase decreased under normoxic and hypoxia( P <0 01). Conclusion Hypoxia induced expression of c fos and c jun genes, which might play an vital role in the early stage of PASMCs proliferation.SOCS3 protein may inhibit proliferation of PASMCs by lowering the tyrosine phosphorylated level of STAT3 protein under hypoxia.

关 键 词：缺氧 MRNA表达 SOCS c-jun c-fos 共转染 肺动脉平滑肌细胞 半定量RT-PCR 负调控因子 增殖 

分 类 号：R346[医药卫生—基础医学]