癌变·畸变·突变 ›› 2009, Vol. 21 ›› Issue (4): 271-275.doi: 10.3969/j.issn.1004-616x.2009.04.006

• 论著 • 上一篇    下一篇

p63和p73的表达与苯并(a)芘致H1299和16HBE细胞DNA损伤的关系

姜 英1;2;饶凯敏1;2;陈 曦1;2;汪 倩1;2;刘爱林1;2;袁 晶1;2;   

  1. 华中科技大学同济医学院公共卫生学院: 1.劳动卫生与环境卫生学系;2.环境与健康教育部重点实验室,武汉 430030
  • 收稿日期:2009-02-19 修回日期:2009-03-06 出版日期:2009-07-30 发布日期:2009-07-30
  • 通讯作者: 袁 晶

Associations between Expressions of p63, p73 and BaP-induced DNA Damage in H1299 and 16HBE Cells

JIANG Ying1;2;RAO Kai-min1;2;CHEN Xi1;2; WANG Qian1;2; LIU Ai-lin1;2;YUAN Jing1;2;   

  1. 1.Department of Occupational and Environmental Health; 2.The MOE Key Laboratory of Environment and Health, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
  • Received:2009-02-19 Revised:2009-03-06 Online:2009-07-30 Published:2009-07-30
  • Contact: YUAN Jing

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摘要： 背景与目的: 研究p63与p73的mRNA表达与BaP致人肺腺癌细胞(H1299)和人支气管上皮细胞(16HBE)DNA损伤的关系、 材料与方法: 分别用不同浓度BaP(8 、16 、32 、64和128 μmol/L)处理H1299和16HBE两种细胞,在4 h和12 h时,使用相应的生化检测试剂盒分别测定细胞裂解液中MDA的水平和SOD、GSH-Px的活性,用qRT-PCR方法测定处理后细胞的p53、p63、p73、mdm2和mdm4的mRNA水平;用Comet实验评价细胞DNA损伤程度、 结果: 16、32和64 μmol/L BaP处理4 h时,两种细胞MDA水平显著性升高,SOD和GSH-Px活性显著性下降(P<0.05)、用BaP处理H1299和16HBE细胞4 h和12 h时均观察到DNA损伤随浓度增加而加重,且呈剂量-效应关系(P<0.01), mdm2、mdm4 mRNA表达水平升高(P<0.01)、不过仅在12 h时p53 基因mRNA表达水平较对照组显著增加(P<0.01)、在4 h和12 h时点,仅在H1299细胞的p63和p73 mRNA表达增加(P<0.05)、 结论: 在BaP致p53缺失的H1299细胞的DNA损伤中,BaP可能通过不依赖p53信号通路激活了p63和p73 mRNA的表达、

关键词: 苯并(a)芘, DNA损伤, p63基因, p73基因

Abstract: BACKGROUND AND AIM: The associations between the changes of p63 and p73 in mRNA levels and the BaP-induced DNA damage in H1299 cells and in 16HBE cells were investigated. MATERIALS AND METHODS: Both H1299 cells and 16HBE cells were treated with BaP at various concentrations (8,16,32,64 and 128 μmol/L) for 4 h and 12 h.At the two time points, the levels of MDA、SOD and GSH-Px were measured using the test kits. The mRNA levels of p53,p63,p73,mdm2 and mdm4 genes were detected by RT-PCR assay. DNA damage in the cells were evaluated by Comet assay. RESULTS: At 4 h,enhanced MDA level was observed (P<0.05). However, levels of SOD and GSH-Px were significantly decreased(P<0.05 for both) in both kinds of cells. After 16HBE and H1299 cells were treated for 4 h and 12 h, DNA damage and mRNA expression levels of mdm2 and mdm4 genes increased in a dose-dependent manner(P<0.01 for all). But enhanced mRNA expression level of p53 was observed only at 12 h(P<0.01). In addition,significant up-regulation of p63 and p73 genes in mRNA levels at the two time points were observed in H1299 cells(P<0.05). CONCLUSION: The BaP-induced DNA damage may be associated with enhanced mRNA levels of p63 and p73 genes in the p53-null H1299 cells.

Key words: benzo(a)pyrene, DNA damage, p63 gene, p73 gene

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