SOCS3基因转染对低氧条件下大鼠肺动脉平滑肌细胞增殖的影响  

Effects of transfected SOCS3 gene on proliferation of rat pulmonary arterial smooth muscle cells under hypoxia

作　　者：白莉[1] 余祖滨[2] 钱桂生[1] 李淑萍[1] 关崧[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,全军呼吸病研究重点实验室,重庆400037 [2]第三军医大学新桥医院胸外科,重庆400037

出　　处：《第三军医大学学报》2004年第20期1802-1805,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目 ( 39970 32 9)~~

摘　　要：目的　探讨SOCS3基因对低氧大鼠肺动脉平滑肌细胞 (PASMCs)增殖的影响。方法　以脂质体为载体将pEFSOCS3和pSV2neo共转染至体外培养的PASMCs ,应用RT PCR和免疫细胞化学法测定转染前后细胞中SOCS3mRNA和蛋白表达。低氧处理PASMCs ,采用Westernblot法检测转染前后常氧 (N)、低氧 6h(H6)、低氧 12h(H12 )PASMCs中STAT3蛋白酪氨酸磷酸化水平变化 ;流式细胞仪、3 H TdR掺入法检测转染前后各时相点细胞增殖情况。结果　RT PCR和免疫细胞化学证实转染后细胞中有SOCS3mRNA和蛋白稳定表达 ;与同时相对照组 (包括未转染组和转染空载体组 )相比 ,SOCS3基因转染组细胞中STAT3蛋白酪氨酸磷酸化水平显著下降 (P <0 0 1) ;流式细胞仪分析显示SOCS3基因转染组细胞各时相点G0 /G1期细胞比例显著增多 (P <0 0 1) ,S +G2 /M期细胞比例显著减少 (P <0 0 1) ;SOCS3基因转染组细胞在常氧和低氧条件下 3 H TdR掺入量显著低于同时相点对照组细胞 (P <0 0 1)。结论　SOCS3蛋白可能通过降低STAT3酪氨酸磷酸化水平抑制PASMCs增殖。Objective To explore the effects of transfected SOCS3 gene on the proliferation of rat pulmonary arterial smooth muscle cells (PASMCs) under hypoxia. Methods PASMCs were cotransfected with pEFSOCS3 and pSV2neo by liposome method. The expressions of SOCS3 mRNA and protein were detected by RT-PCR and immunocytochemistry, respectively. Tyrosine-phosphorylated Stat3 level was measured by Western blotting. Flow cytometric DNA analysis and 3H-TdR incorporation was used to detect the cell proliferation. Results The SOCS3 expression was confirmed by RT-PCR and Western blotting in PASMCs transfected with SOCS3 gene, respectively. Compared with that in the control group (non-transfected and pEF and psv2neo cotransfected A549) at the same hour under hypoxia, tyrosine-phosphorylated STAT3 level decreased greatly in PASMCs transfected with SOCS3 gene (P<0.01); the cells in PASMCs transfected with SOCS3 gene at G 1/G 0 phase increased and those at G 2/M+S phase decreased significantly under nomoxia and hypoxia (P<0.01). 3H-TdR incorporation in PASMCs transfected with SOCS3 gene was significantly lower than that in the control group under nomoxia and hypoxia (P<0.01). Conclusion SOCS3 protein may inhibit the proliferation of PASMCs by down-regulating the tyrosine-phosphorylated level of STAT3 protein.

关 键 词：低氧 细胞因子 信号传导 肺动脉 平滑肌细胞 增殖 

分 类 号：R322.121[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学]