石英通过丝裂素活化蛋白激酶通路引起人胚肺成纤维细胞周期的改变  

Silica induce cell cycle changes by mitogen-activated protein kinases pathway

作　　者：贾效伟[1] 刘秉慈[1] 叶萌[1] 刘海峰[2] 张凤梅[3] 

机构地区：[1]中国疾病预防控制中心职业卫生与中毒控制所,北京100050 [2]唐山市疾病预防控制中心 [3]山东大学公共卫生学院

出　　处：《中华劳动卫生职业病杂志》2011年第7期487-491,共5页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金资助项目（30972449,30671747）.

摘　　要：目的 探讨在石英刺激的人胚肺成纤维细胞（human embryo lung fibroblasts,HELF）中细胞外信号调节蛋白激酶（ERK）、应激活化蛋白激酶（JNK）,细胞周期蛋白DI（cyclin D1）通路在石英诱导的细胞周期改变中的作用.方法 建立稳定转染转录因子AP-1荧光素酶报告基冈质粒的HELF系（HELF-AP-1）及AP-1荧光素酶报告基因质粒与丝裂素活化蛋白激酶（MAPK）显性失活突变体质粒（DN-ERK、DN-JNK和DN-p38）分别共转染的HELF系（简称DN-ERK、DN-JNK和DN-p38）.将HELF-AP-1、DN-ERK、DN-JNK和DN-p38细胞分为对照组和石英组（共8组）,各对照组不加任何处理,石英组用200 μg/ml石英处理HELF细胞24 h.用免疫印迹（Western blot）法和免疫荧光法检测cyclin D1、细胞周期蛋白依赖激酶4（CDK4）和E2F-4蛋白表达;采用显性失活突变体技术验证MAPKs信号转导通路的上下游关系及其与细胞周期的关系;用流式细胞术检测细胞周期变化.结果 HELF-AP-1＋石英组G1期细胞所占比例下降,S期细胞所占比例升高,与HELF-AP-1对照组的差异有统计学意义（P＜0.05）.抑制ERK或JNK表达后,与HELF-AP-1对照组比较,DN-ERK＋石英组和DN-JNK＋石英组G1期细胞百分比无明显变化.抑制p38后,DN-p38＋石英组G1期细胞百分率明显下降,与HELF-AP-1对照组的差异有统计学意义（P＜0.05）.与HELF-AP-1石英组比较,DN-ERK＋石英组和DN-JNK＋石英组CDK4表达降低和E2F-4表达增多,而DN-p38＋石英组CDK4表达和E2F-4表达没有改变.与HELF-AP-1＋石英组比较,DN-ERK＋石英组和DN-JNK＋石英组cyclin D1表达降低,而DN-p38＋石英组cyclin D1没有改变.结论 ERK和JNK通过cyclin D1和CDK4介导石英诱导的HELF的细胞周期改变,而石英诱导的细胞周期改变与p38无关.Objective To investigate the roles of mitogen-activated protein kinases （MAPK） on silica-induced cell cycle changes. Methods After cells were treated with 200 μg/ml silica, Western blot and Immunofluorescence assays were utilized to detect the expression of cyclin D1, CDK4 and E2F-4, Flow cytometry was used to detect cell cycle progression, the dominant negative mutants techniques were used to investigate silica-induced signal pathway and the effects of which in silica-induced cell cycle changes. Results After cells were exposed to 200 μg/ml silica 24 h, the results of present study showed the proportion of cells in G1 phases was decreased. Silica-induced cell cycle alternation was markedly impaired by stable expression of a dominant negative mutants of ERK or JNK, but not p38. It was found that ERK and JNK were involved in silica-induced cyclin D1 and CDK4 overexpression and the decreased expression of E2F-4. Conclusion ERK and JNK, but not p38, mediated silica-induced cell cycle changes in human embryo lung fibroblasts.

关 键 词：石英 成纤维细胞 细胞周期 信号传递 

分 类 号：R686[医药卫生—骨科学]