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

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

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36h，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48h使大部分细胞处于静止期后，加入SiO2刺激6h的AM培养上清（S6组），用反转录-聚合酶链反应（RT—PCR）及Westemblot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7h，观察c—mycmRNA和蛋白表达的变化。结果HEFB在静止状态时c—mycmRNA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—mycmRNA及蛋白的表达，以AM培养12h的上清作用最明显，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—mycmRNA和蛋白的表达，但作用高峰提前，以SiO2刺激6h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6h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HEFBc—myc基因的表达。

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（30 μg/ml） for 1,2,6,12,24 and 36 h; （2）control：treated for the same time without SiO2. Fibroblasts were cultured with different AMs supernatants for 2 h or 7 h respectively. The expression of c-myc mRNA was determined by RT-PCR and protein by Western Blot. Results 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 SiO2 （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.