矽肺患者肺泡巨噬细胞培养上清对人胚肺成纤维细胞c-myc基因表达的影响

目的 探讨矽肺患者肺泡巨噬细胞(AM)培养上清对人胚肺成纤维细胞(HEFB)c-myc基因表达的影响.方法 AM分为加尘组(SiO2,30μg/ml)和对照组,培养1、2、6、12、24和36 h,收集培养上清.Ⅲ代HEFB用质量分数为0.5%的血清培养液培养48 h使大部分细胞处于静止期后,加人SiO2刺激6 h的AM培养上清(S6组),用反转录一聚合酶链反应(RT-PCR)及Western blot方法观察c-mycmRNA和蛋白的时程变化.然后将各组AM上清与静止状态的HEFB孵育2和7 h,观察c-myc mRNA和蛋白表达的变化.结果 HEFB在静止状态时c-myc mRNA和蛋白的表达为0,在对照组中,培养不同时间的AM上清刺激了c-myc mRNA及蛋白的表达,以AM培养12 h的卜清作用最明显,比值分别为0.749±0.088和0.759±0.101.加尘组中各上清也刺激了c-myc mRNA和蛋白的表达,但作用高峰提前,以SiO2刺激6 h的上清作用最明显,比值分别0.982±0.147和0.978±0.141.与同期对照相比,SiO2刺激1、2和6 h的AM上清诱导mRNA和蛋白表达增多,差异有统计学意义(P＜0.05或P＜0.01).结论 SiO2激活的AM培养上清可促进HEFB c-myc基因的表达.

Effect of silicosis alveolar macrophages on expression of c-myc in human embryo lung fibroblast in vitro

Objective To study the effect of silicosis alveolar macrophages(AM)restimulated by SiO2 on expression of c-myc oncogene in human embryo lung fibroblasts.Methods The brochoalveolar lavage of silicosis patients was collected.AMs were divided into 2 groups:(1)SiO2:AMS were stimulated with SiO2(30μg/ml)for 1,2,6,12,24 and 36 h;(2)control:treated for the same time without SiO2.Fibroblasts were cul tured with different AMS supematants for 2 h or 7 h respectively.The expression of c-myc mRNA was determined bv RT-PCR and protein bv Western Blot.Resuits There was no c-myc expression when fibroblasts were static.The supernatants in the S6 group stimulated expression of c-myc mRNA and protein,with the peak expression at 2 h and 7 h respectively.In the control group,AMS supernatants cultured in different time stimulated expression of c-myc mRNA and protein with the most evident expression at 12 h.The ratios were 0.749±0.088 and 0.759±0.101 respectively.Compared with control in the same period,c-myc mRNA and protein expression were significantly stronger treated with the supernatants in which AMs were stimulated for 1 h.2 h and 6 h by SiO,(P＜0.05 or P＜0.01).Conclusion AMs stimulated with SiO2 has the ability to induce c-myc oncogene expression in human embryo lung fibroblasts.