全反式视黄酸阻断苯并(a)芘致成纤维细胞周期改变的通路  

Inhibition of the pathway of benzo(a)pyrene-induced cell cycle changes by all-trans retinoic acid in lung fibroblast

作　　者：贾效伟[1] 刘秉慈[1] 史香林[2] 高艾[1] 尤宝荣[1] 叶萌[1] 沈福海[1] 杜宏举[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]中国科学院上海生命科学研究院营养科学研究所

出　　处：《中华劳动卫生职业病杂志》2005年第5期329-332,共4页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金资助项目(30028019;30371206);"973"国家重点基础研究发展规划资助项目(2002CB512906);香港泰山公德会资助项目

摘　　要：目的观察全反式视黄酸(ATRA)逆转苯并(a)芘诱导的人胚肺成纤维细胞(HELF)细胞周期紊乱过程中细胞周期素D1(cyclinD1)、细胞周期素依赖性激酶K4(CDK4)、转录因子E2F1和E2F4的作用。方法用0.1μmol/LATRA预处理细胞后,再用2μmol/L苯并(a)芘作用细胞,用Westernblotting方法测定其蛋白表达水平;建立稳定转染了反义cyclinD1质粒和反义CDK4质粒的细胞系,应用反义技术证明上下游关系;用流式细胞技术测定细胞周期。结果2μmol/L苯并(a)芘作用于HELF24h后,cyclinD1、E2F1蛋白表达水平明显增高,CDK4和E2F4蛋白表达水平无明显变化;用0.1μmol/LATRA预处理24h后,cyclinD1、E2F1蛋白表达水平明显下降;与相同作用的对照组相比,苯并(a)芘刺激反义cyclinD1或反义CDK4细胞后E2F1表达增加的幅度明显降低;与相同作用的对照组相比ATRA预处理的反义cyclinD1或反义CDK4细胞中,苯并(a)芘诱导的E2F1过表达水平降低的幅度明显降低;流式细胞术测定结果显示,苯并(a)芘诱导细胞从G1期进入S期,ATRA通过聚积G1期细胞而阻滞苯并(a)芘诱导的细胞周期进程。结论ATRA通过cyclinD1/E2F1途径阻断苯并(a)芘诱导的HELF的细胞周期改变。Objective To investigate the reverse effect of all-trans retinoic acid（ATRA） on Benzo（a） pyrene （B（a）P）-induced cyclin D1 ,CDK4,E2F-1 and E2F-4 expression and cell cycle progression in human embryo lung fibroblast（HELF）. Methods After HELF cells was treated with ATRA,they were exposed to 2μmoL/L of B（a）P. Western blotting was employed to detect protein expression level;the RNA transfection techniques was used to investigate ATRA-induced signal pathway;flow cytometry was used to detect cell cycle progression. Result After treatment with 2μmoL/L B（a）P for 24 h,the expression of cyclin D1 and E2F-1 were both increased signitlcantly in HELF;the expression of E2F-4 and CDK4 were not changed markedly;pretreatment with 0.1μmoL/L ATRA for 24 h could efficiently decrease B（a） P-induced overexpression of cyclin D1 and E2F-1 ;stimulation to antisense cyclin D1 or antisense CDK4 by B（a） P could significantly impair E2F-1 up-regulation; pretreatment with ATRA, cells with antisense cyclin D1 or antisense CDK4 showed a less decrease in B（a）P-induced overexpression of E2F-1 compared to similarly treated control cells;flow cytometry analysis showed B（a） P promoted cell cycle progression from G1 phase to S phase,while pretreatment with ATRA could inhibit B（a）P-induced cell cycle progression by an accumulation of cells in the GI phase. Conclusion ATRA could block B（a）P-induced cell cycle pro- motion through cyclin D1/E2F-1 pathway in HELF.

关 键 词：全反式视黄酸 苯并(A)芘 细胞周期素D1 转录因子 细胞周期 细胞周期素依赖性激酶 苯并(A)芘 人胚肺成纤维细胞 全反式视黄酸 转录因子E2F-1 阻断 cyclin 蛋白表达水平 BLOTTING 

分 类 号：R73-3[医药卫生—肿瘤]