苯并(a)芘作用下人胚肺细胞JWA基因的表达及其与DNA损伤修复的关系

目的研究苯并(a)芘诱导人胚肺(ccc-HPF-1)细胞DNA损伤修复中基因JWA和热休克蛋白(hsp70)的表达规律,探讨JWA基因参与细胞DNA损伤修复的作用和可能机制。方法建立ccc-HPF-1细胞DNA损伤模型,在S9活化状态下分别以10～100μmol／L苯并(a)芘处理细胞3h收获细胞或再恢复24h,用碱性单细胞凝胶电泳鉴定DNA损伤和修复情况,免疫印迹方法检测JWA和hsp70的表达水平。结果在DNA损伤模型中,苯并(a)芘活跃地调节JWA的表达,50μmol／L和100μmol／L处理组JWA和hsp70明显上调,在恢复期内10～100μmol／L处理组两者都处于较高的表达水平并且JWA和hsp70的表达规律几乎完全一致。结论JWA是一个有效的环境应答基因,活跃地参与细胞应答与DNA切除修复有关的信号通路。

Relationship between JWA expression and DNA damage-repair in human embryonic lung cells by benzo(a) pyrene

OBJECTIVE: To investigate the expressions of JWA gene and heat shock protein 70 (hsp70) in human embryonic lung (cccHPF-1) cells after exposure to activated benzo(a)pyrene (B(a)P), and to explore the role and the possible mechanism of JWA gene involved in B(a)P-induced DNA damage and repair. METHODS: The models of DNA damage of ccc-HPF-1 cells were established by treatment of cells with B(a)P plus S9 at 10 to 100 micromol/L for 3 hours with or without 24 hours recovery for DNA repairing. The DNA damage was detected by single cell gel electrophoresis (SCGE) assay (comet assay). And the immuno-blotting assay was used for detecting expressions of JWA and hsp70. RESULTS: JWA expression was actively modulated by B(a)P exposure. The expressions of both JWA and hsp70 were increased greatly at 50 micromol/L and 100 micromol/L B(a)P treated cells and maintained at over expressed levels treated by 10-100 micromol/L B(a)P during the restored time. In addition, JWA expression pattern was similar to that of hsp70. CONCLUSION: JWA is determined in this study by its functioning as an effective environmental responsive gene and actively participating in the signal pathways of DNA damage and repair which might be associated with excision repair.