论文部分内容阅读
目的:探讨SOCS3基因对缺氧大鼠肺动脉平滑肌细胞(PASMCS)原癌基因C-MYC MRNA表达及细胞增殖的影响。方法:以脂质体为载体将PEFSOCS3和PSV2NEO共转染至体外培养的PASMCS,G418筛选阳性克隆,应用免疫细胞化学法测定转染前后细胞中SOCS3蛋白表达;缺氧处理转染组和对照组PASMCS,采用半定量RT-PCR检测转染前后常氧组、缺氧培养2 H、6 H、12 H、16 H、24 H细胞C-MYC MRNA表达水平变化;[3H]-TDR掺入法检测转染前后上述各时点细胞增殖情况。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结果显示,SOCS3基因转染组细胞C-MYC MRNA表达水平在缺氧各时点均显著低于同时点对照组细胞(P<0．01);SOCS3 基因转染组细胞在常氧和缺氧各时点[3H]-TDR掺入量显著低于对照组细胞(P<0．01)。结论:SOCS3蛋白可能通过下调C-MYE基因表达从而抑制缺氧诱导的PASMCS增殖。
Objective: To investigate the effect of SOCS3 gene on the expression of proto-oncogene C-MYC MRNA and cell proliferation in pulmonary artery smooth muscle cells (PASMCS) of hypoxic rats. METHODS: PEFSOCS3 and PSV2NEO were co-transfected into PASMCS with lipofectin as carrier and positive clones were screened with G418. The expression of SOCS3 protein in cells before and after transfection was determined by immunocytochemistry. PASMCS. The expression of C-MYC MRNA was detected by semi-quantitative RT-PCR in normoxia group and H 2 H, 6 H, 12 H, 16 H, 24 H cells under hypoxia. [3H] Detection of cell proliferation at various time points before and after transfection. Results: Immunocytochemistry confirmed the stable expression of SOCS3 in transfected cells. Semi-quantitative RT-PCR results showed that the expression level of C-MYC mRNA in SOCS3 transfected group was significantly lower than that in control group at each time point (P <0.01). The amount of [3H] -TDR incorporation in SOCS3 transfection group was significantly lower than that in control cells at each time of normoxia and hypoxia (P <0.01). Conclusion: SOCS3 protein may inhibit hypoxia-induced PASMCS proliferation by down-regulating C-MYE gene expression.