苯并(a)芘对H1299细胞CYP1A1酶活力和相关基因表达的影响  

CYP1A1 activity,and expressions of CYP1A1 and related genes in H1299 cells treated with benzo(a)pyrene

作　　者：姜英[1,2] 陈曦[1,2] 饶凯敏[1,2] 杨光涛[1] 汪倩[1,2] 刘爱林[1,2] 袁晶[1,2] 

机构地区：[1]华中科技大学同济医学院公共卫生学院劳动卫生与环境卫生学系,湖北武汉430030 [2]环境与健康教育部重点实验室

出　　处：《毒理学杂志》2010年第4期253-257,共5页Journal of Toxicology

基　　金：国家自然科学基金(30671735)

摘　　要：目的研究BaP对人肺腺癌细胞(H1299)CYP1A1酶活力和CYP1A1及其相关基因表达的影响。方法用不同浓度BaP(8、16、32、64和128μmol/L)处理H1299细胞24 h。实时荧光定量PCR方法检测H1299细胞中CYP1A1、HSP90、AhR和ARNT基因的mRNA表达水平;表面等离子共振技术(surface plasmon resonance,SPR)检测CYP1A1和HSP90蛋白表达水平;荧光分光光度法测定CYP1A1酶活力(7-ethoxyresorufin-o-deethylase,EROD)。结果与溶剂对照组相比,CYP1A1和ARNT基因mRNA水平在BaP为16和64μmol/L时有统计学意义(P<0.01),AhR和HSP90在各染毒组中均有统计学意义(P<0.05)。在16、32和64μmol/L的BaP染毒组中,CYP1A1和HSP90蛋白水平高于对照组。在16、32、64和128μmol/L的BaP染毒组中均可诱导CYP1A1酶活力显著性增强(P<0.01)。结论该试验条件下,在H1299细胞,BaP可通过诱导CYP1A1与其上游转录因子AhR和ARNT的mRNA以及蛋白表达而增强CYP1A1酶活力。Objective To investigate the CYP1A1 activity and expressions of CYP1A1,AhR,ARNT and HSP90 genes in H1299 cells treated with BaP.Methods H1299 cells were treated with BaP at indicated concentrations（8,16,32,64 and 128 μmol/L）.After the treatment of 24 h,the mRNA levels of CYP1A1,HSP90,AhR and ARNT genes were detected by RT-qPCR assay;the expressions of CYP1A1 and HSP90 proteins were detected using Surface Plasmon Resonance（SPR） technique;the CYP1A1 activity was also detected by using fluorescence spectrophotometry.Results These results show that mRNA expressions were significantly enhanced for CYP1A1 and ARNT genes in Bap-treated groups at concentrations of 16 and 64μmol /L（P〈 0.01）,for AhR and Hsp90 genes in all BaP-treated groups（P〈 0.05）.We also found that higher protein expressions of CYP1A1 and HSP90 in BaP-treated groups（16,32 and 64 μmol /L）.Additionally,CYP1A1 activities were significantly increased in the BaP-treated groups except of 8 μmol /L（P〈 0.01）.Conclusion BaP induced CYP1A1 activity by the increased mRNA expres-sions of CYP1A1 and the upstream transcriptional factors including AhR and ARNT.

关 键 词：苯并(A)芘 表面等离子体共振技术 CYP1A1 AHR ARNT 

分 类 号：R114[医药卫生—卫生毒理学]