Ye Meng, Liu Bing-ci, Jia Xiao-wei, Gao Ai, Jiao Shi, Zhang Feng-mei
National Institute of Occupation Health and Poison Control, Chinese Center for Disease Control and Prevention, Beijing 100050, China.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2008 Feb;26(2):72-6.
To investigate the roles of activated protein 1 (AP-1) in cell cycle changes on human embryo lung fibroblasts (HELF) induced by benzo (a) pyrene [B (a) P], and relationships between AP-1 and cyclin D1/CDK4-E2F-1/4.
Cells transfected with AP-1 luciferase reporter plasmid (AP-H) were cultured with serum-free RPMI1640 for 48 h, and treated with 2 micromol/L B (a) P for 24 h. AP-1 relative activity was detected by luciferase assay. Changes of cell cycle and the expression of cyclin D1, CDK4 and E2F-1/4 were checked using the flow cytometer and Western blot assay.
After B (a) P was treated for 24 h, the ratio of G1 phase cells (71 +/- 2)% was decreased to (48 +/- 3)% (P < 0.05), and an increase was observed in the ratio of S phase. AP-1 activity and cyclin D1/E2F-1 expression were increased significantly, but CDK4/E2F-4 expression did not change after B (a) P treatment. When AP-1 activity was inhibited by curcumin, decreases of G1 phase in response to B (a) P treatment were blocked, and overexpression of cyclin D1/E2F-1 was attenuated, but CDK4/E2F-4 expression was not changed significantly.
AP-1 is involved in B (a) P induced cell cycle changes, and is the upstream signals of cyclin D1/E2F-1, but not CDK4/E2F-4.
探讨活化蛋白1（AP-1）在苯并（a）芘[B（a）P]诱导人胚肺成纤维细胞（HELF）细胞周期变化中的作用，以及AP-1与细胞周期蛋白D1/细胞周期蛋白依赖性激酶4（CDK4）-E2F-1/4之间的关系。
将转染AP-1荧光素酶报告质粒（AP-H）的细胞用无血清RPMI1640培养48小时，并用2微摩尔/升B（a）P处理24小时。通过荧光素酶测定检测AP-1相对活性。使用流式细胞仪和蛋白质免疫印迹法检测细胞周期变化以及细胞周期蛋白D1、CDK4和E2F-1/4的表达。
B（a）P处理24小时后，G1期细胞比例由（71±2）%降至（48±3）%（P<0.05），S期比例增加。B（a）P处理后AP-1活性及细胞周期蛋白D1/E2F-1表达显著增加，但CDK4/E2F-4表达未改变。当用姜黄素抑制AP-1活性时，B（a）P处理引起的G1期降低被阻断，细胞周期蛋白D1/E2F-1的过表达减弱，但CDK4/E2F-4表达无明显变化。
AP-1参与B（a）P诱导的细胞周期变化，是细胞周期蛋白D1/E2F-1的上游信号，但不是CDK4/E2F-4的上游信号。
