癌变·畸变·突变 ›› 2009, Vol. 21 ›› Issue (1): 42-045.doi: 10.3969/j.issn.1004-616x.2009.01.010

• 论著 • 上一篇    下一篇

苯并芘对HELF细胞周期及相关基因表达的影响

王智琴1,2,齐以涛1,杨 迪1,陈 倩1,周宗灿3,王 捷2,梁 震2,肖希龙1   

  1. 1.中国农业大学动物医学院药理与毒理教研室,北京 100193;2.沈阳化工研究院安评中心,辽宁 沈阳 110021;3.北京大学医学部公共卫生学院毒理教研室, 北京 100083
  • 收稿日期:2008-08-28 修回日期:2008-10-07 出版日期:2009-01-30 发布日期:2009-01-30

The Effect of Benzo[a]pyrene Exposure on Cell Cycle and Related Genes in HELF Cells

WANG Zhi-qin1,2, QI Yi-tao1, YANG Di1, CHEN Qian1, ZHOU Zong-can3, WANG Jie2, LIANG Zhen2, XIAO Xi-long1,   

  1. 1.Department of Pharmacology and Toxicology, College of Veterinary Medicine, China Agricultural University, Beijing 100193; 2. National (Shenyang) New Drug Evaluation Center, Chemical and Industrial Research Institute, Shenyang 110021; 3. Department of Toxicology,Health Science Center, Peking University, Beijing 100083
  • Received:2008-08-28 Revised:2008-10-07 Online:2009-01-30 Published:2009-01-30

摘要: 背景与目的: 探讨苯并芘(benzo[a]pyrene, BaP)影响人胚肺成纤维细胞(HELF)周期的途径和作用机制。 材料与方法: 用5 μmol/L的BaP处理HELF细胞后,采用细胞计数,流式细胞仪检测BaP对细胞生长和周期的影响,用PCR和Western-blotting检测BaP对细胞周期相关基因mRNA和蛋白的影响。 结果: BaP处理HELF细胞后,显著促进细胞的生长,在处理后第8 d时细胞数目约增加50%(P<0.01),细胞体积增大,促进细胞由G1期向S期和G2/M期的转换。 mRNA和蛋白检测结果显示:BaP处理细胞后可以促进p53、cyclin D1、CDK2、CDK4和p21 mRNA及蛋白的表达(P<0.05或P<0.01),抑制cyclin E mRNA及蛋白的表达(P均<0.01),而对cyclin A和p27 mRNA及蛋白表达没有明显影响(P均>0.05)。 结论: BaP对HELF细胞周期的调控逃脱了p53-p21的关卡作用,而通过诱导cyclin D1、CDK2和CDK4的表达来加快细胞周期的进程,该途径为cyclin A和p27非依赖型。

关键词: 苯并芘, 细胞周期, 细胞周期蛋白, 细胞周期蛋白依赖激酶, CKIs, 人胚肺成纤维细胞

Abstract: BACKGROUND AND AIM: To examine the pathway and mechanisms of benzo[a]pyrene(BaP) effects on cell cycle. MATERIALS AND METHODS: We administratered BaP to human embryonic lung fibroblasts(HELF) and monitored the changes in cell growth by cell counting, the cell size and cell cycle distribution by FACScan flow cytometer, and the mRNA and protein expression levels of cell cycle genes by RT-PCR and Western blotting. RESULTS: BaP promoted HELF cell growth, enlarged cell size, enhanced the transition of G1 phase to S and G2/M phase. The mRNA and protein expression levels of p53-related genes were detected indicating that BaP up-regulated the p53 mRNA and protein expression, and increased the expressions of cyclin D1, CDK2, CDK4 and p21, while decreasing the expresson of cyclin E. CONCLUSION: BaP affected cell cycle progression via the expressions of cyclin D1, CDK2 and CDK4, escaping from the p53-p21 mediated G1 checkpoint and was cyclinA-p27 independent.

Key words: BaP, cell cycle, cyclin, Cdk, CKIs, HELF