cyclinD1/E2F通路在苯并(a)芘引起人胚肺成纤维细胞周期改变中的作用  

cyclinD1/E2F pathways involved in cell cycle changes of human embryo lung fibroblasts induced by benzo(a)pyrene

作　　者：叶萌[1] 刘秉慈[1] 史香林[2] 尤宝荣[1] 杜宏举[1] 贾效伟[1] 沈福海[1] 

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]中国科学院上海营养科学研究所

出　　处：《中华劳动卫生职业病杂志》2006年第2期67-71,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(30371206;30440420593);"973"国家重点基础研究发展规划项目(2002CB512905);美国PhilipMorrisUSAIncandbyPhilipMorrisInternational项目

摘　　要：目的探讨细胞周期蛋白D1(cyclin D1)/细胞周期蛋白依赖激酶4(CDK4)-E2F-1/4通路在苯并(a)芘[B(a)P]引起的人胚肺成纤维细胞(HELF)周期改变中的作用。方法分别用2μmol/L的B(a)P作用HELF 24 h和100μmol/L B(a)P作用3次,每次24 h,细胞具有转化细胞的部分特征(T-HELF)。用转染反义cyclin D1和CDK4质粒的HELF(A-D1,A-K4)和T-HELF(T-A-D1,T-A-K4)研究cy-clin D1/CDK4与E2F-1/4的上下游关系。用流式细胞仪和Western blot法检测细胞周期、cyclin D1、CDK4和E2F-1/4蛋白表达的改变。结果2μmol/L B(a)P作用24 h HELF中G1期细胞数明显减少。在A-D1和A-K4的HELF中,cyclin D1和CDK4表达的抑制阻断了由B(a)P引起的细胞周期从G1期进入S期的变化。B(a)P刺激后HELF中cyclin D1和E2F-1的表达明显增加。在A-D1中,E2F-1的高表达被抑制。B(a)P作用A-K4细胞后,CDK4表达明显升高,E2F-4表达明显降低。T-A-D1和T-A-K4的T-HELF多数停留在G1期。与HELF相比,T-HELF中cyclin D1的表达明显增加;与T-HELF相比,T-A-D1和T-A-K4中E2F-4表达明显增加。结论在2μmol/L B(a)P作用的HELF中,B(a)P通过cyclin D1/CDK4-E2F-1/4信号转导通路引起细胞周期的改变。在T-HELF中,B(a)P通过cyclin D1/CDK4-E2F-4信号转导通路引起细胞周期的改变。Objective To investigate the roles of cyclin D1/CDK4-E2F-1/4 pathway in cell cycle changes of human embryo lung fibroblasts （HELF） induced by two different benzo（a） pyrene [ B（a） P] treatment models. Methotls Two B（a）P treatment models： HELF were treated by 2μmol/L B（a）P for 24 hours; HELF were treated by 100 μmol/L B（a）P three times 24 hours each and provide with some characteristics of transformed cells （THELF）. Changes of cell cycle and the expression of cyclin D1, CDK4 and E2F-1/4 were checked using the flow cytomter and Western bolt analysis. Results After 24 hours 2μmL/L B（a）P treatment,the HELFs in the G1 phase was decreased. In HELF transfected with antisense cyclin D1 （A-D1） and antisense CDK4 （A-K4）, the expression of cyclin D1 and CDK4 blocked the cell cycle changes from the G1 phase to the S phase induced by B（a）P. The overexpression of cyclin D1 and E2F-1 in HELF was induced by B（a）P. The E2F-1 overexpression in A-D1 induced B（a） P was inhibited. The E2F-4 expression was decreased and the CDK4 expression was increased significantly in A-K4 after B（a）P treatment.Most of T-HELF Iransfected with antisense cyclin D1 （T-A-D1） and antisense CDK4 （T-A-K4） were retained in Ga phase. The cyclin D1 expression in T-HELF＊was increased significantly compared with that in HELF.The E2F-4 expression in T-A-D1 and T-A-K4 was increased signiticantly compared with that in T-HFZF. Conclusion B（a） P induces the cell cycle changes through cyclin D1/CDK4-E2F-1/4 pathway in HELF treated by 2μmol/L B（a）P while it induces cell cycle changes through cyclin D1/CDK4-E2F-4 pathway in T-HELF.

关 键 词：苯并(A)芘 基因 细胞周期蛋白D1 转录因子 细胞周期 

分 类 号：R131[医药卫生—劳动卫生]