中文摘要：

目的 探讨铜蓝蛋白（Ceruloplasmin,Cp）在石英诱导的人胚肺成纤维细胞（HELF）中PI3K/PTEN信号通路改变中的作用.方法 建立稳定转染空载体pGenesil1.1的HELF细胞及pGenesil 1.1与shRNA PTEN质粒共转染的HELF细胞（简称PT）,复苏转染p85显性失活突变体质粒的HELF细胞（简称DN-p85）.实验中石英浓度为100 μg/ml,Cp浓度为10、20、30 μg/ml,Cp在石英作用1h后加入.在HELF、PT和DN-p85 3个不同细胞系中,分别设立对照组、石英染毒组、石英＋不同浓度Cp组.用噻唑蓝（MTT）比色法观察分别抑制PTEN和p85后,Cp对石英诱导的HELF细胞增殖的影响.蛋白免疫印迹（Western blot）实验检测PTEN、p85、p110、AKT及AKT308位点和AKT473位点磷酸化水平、总ERK、JNK及其磷酸化水平的变化.结果 抑制PTEN后,100 μgml石英与30 μg/ml Cp共同诱导的p85蛋白水平增加被明显抑制（P＜0.05）,p110蛋白水平无变化（P＞0.05）.抑制p85后,石英以及石英＋不同浓度Cp诱导的细胞增殖加快被抑制;100 μg/ml石英与30 μg/ml Cp共同诱导的AKT308位及AKT473位磷酸化水平、总ERK、磷酸化ERK和磷酸化JNK水平的增加低于相应的对照组（P＜0.05）,但是总JNK水平的变化没有统计学差异（P＞0.05）.结论 Cp通过PI3K/AKT/MAPK信号通路参与石英诱导的HELF细胞增殖,PTEN可以影响PI3K调节亚单位p85蛋白水平.

英文摘要：

Objective To investigate the roles of ceruloplasmin （Cp） in PI3K/PTEN cell signaling pathway change in human embryonic lung fibroblasts （HELFs） induced by silica.Methods HELFs transfected with pGenesil1.1 plasmid and pGenesil1.1 with PTEN shRNA （PT） plasmid were successfully established.100 μg/ml silica and different concentrations of Cp （10、20、30 μg/ml） were used in this experiment and Cp were treated cells after exposed to silica for 1 h.Three different cell lines （including HELFs,PT and cells were transfected with p85 dominant negative mutant plasmid （DN-p85）） were divided into control groups,silica groups and silica＋different concentrations of Cp groups.MTT assay was used to dctect the effects of Cp on silica-induced cell proliferation after inhibiting PTEN and p85.When supressing the expression of PTEN and p85,western blot assay was performed to detect the levels of p85,pll0,AKT308,AKT473 and ERK,JNK and their phosphorylated levels.Results After inhibition of PTEN,the high levels of p85 induced by 100 μg/ml silica with 30 μg/ml Cp were markedly decreased （P〈0.05）.When suppressing p85,the increased cell proliferation was not observed.And the high levels of AKT308,AKT473,ERK and phosphorylated JNK and ERK stimulated by 100 μg/ml silica with 30 μg/ml Cp were decreased （P〈0.05）.Conclusion Cp could further strengthened silica-induced cell proliferation by PI3K/AKT/MAPK cell signaling pathway,of which the level of p85 was regulated by PTEN.