PM2.5对支气管上皮细胞DNA损伤作用研究  

PM2.5 on DNA damage in human bronchial epithelial cells

作　　者：王冰玉 郑凯 徐新云[2] 谢红卫[1] 于军晖[1] 龙鼎新[1] WANG Bingyu;ZHENG Kai;XU Xinyun;XIE Hongwei;YU Junhui;LONG Dingxin(School of Public Health,University of South China,Hengyang 421001,Hunan;Institute of Environment and Health,Shenzhen Center for Disease Control and Prevention,Shenzhen 518055,Guangdong,China)

机构地区：[1]南华大学公共卫生学院,湖南衡阳421001 [2]深圳市疾病预防控制中心环境与健康所,广东深圳518055

出　　处：《癌变．畸变．突变》2019年第6期469-473,497,共6页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：深圳市科技研发基础研究学科布局项目(JCYJ20170413101713324)

摘　　要：目的:探讨大气细颗粒物(PM2.5)染毒对人支气管上皮细胞(HBE)DNA损伤的作用。方法:分别用8、20、50μg/mL的PM2.5水溶液染毒HBE细胞24 h后,单细胞凝胶电泳实验(SCGE)检测DNA损伤情况。10和50μg/L的PM2.5水溶液染毒HBE细胞,以未染毒细胞作为阴性对照组,10μmol/L的Cr6+水溶液为阳性对照组,实时荧光定量PCR(qPCR)检测DNA损伤修复基因hOGG1、hMTH1的mRNA表达水平的变化,Western blot检测hOGG1、hMTH1蛋白表达变化。结果:单细胞凝胶电泳检测8、20和50μg/mL PM2.5水溶液染毒组HBE细胞的尾部DNA含量、尾长、尾距较阴性对照组明显增加(P<0.05或P<0.01)。qPCR结果显示,与阴性对照组比较,HBE细胞hOGG1 mRNA表达水平在10和50μg/mL PM2.5水溶液染毒以及阳性对照Cr6+水溶液染毒后分别升高75.0%、132.0%、214.0%;hMTH1 mRNA分别升高61.0%、144.0%、75.0%。Western blot结果显示,与阴性对照组比较,H BE细胞hOGG1蛋白表达水平在10和50μg/mL PM2.5水溶液染毒以及Cr6+水溶液染毒后分别升高47.6%、64.0%、47.0%;hMTH1蛋白分别升高20.5%、49.8%、20.9%。结论:PM2.5水溶液染毒对HBE细胞DNA具有明显的损伤作用,并引起HBE细胞DNA损伤修复基因hOGG1、hMTH1表达水平升高。OBJECTIVE:To investigate the effect of PM2.5 on DNA damage in human bronchial epithelial cells(HBE).METHODS:HBE cells were exposed to PM2.5(0,8,20,50μg/mL)for 24 h and DNA damage was detected using the single cell gel electrophoresis(SCGE)assay.For another investigation,cells were treated with 10 and 50μg/mL,untreated cells were used as negative controland cells treated with 10μmol/L Cr6+were used as positive control.mRNA expressions of DNA damage repair genes,including hOGG1 and hMTH1,were detected using real-time quantitative PCR(qPCR)and protein expressions using Western blot.RESULTS:SCGE data show that the tail DNA content,tail length,tail distance significantly increased in the treated groups compared with the control group(P<0.05 or P<0.01).The qPCR data show that,compared with the control group,expression of hOGG1 mRNA increased by 75.0%,132.0%,214.0%after exposure to PM2.5 at 10,50μg/mL and positive control Cr6+,respectively.Expression of hMTH1 mRNA increased by 61.0%,144.0%and 75.0%,respectively.The Western blot data show that,compared with the control group,expression of hOGG1 protein increased by 47.6%,64.0%,47.0%after exposure to PM2.5 at 10,50μg/mL and positive control Cr6+,respectively.Expression of hMTH1 protein increased by 20.5%,49.8%,20.9%,respectively.CONCLUSION:Our investigation shows that PM2.5 induced DNA strand breaks which caused dose-dependent increase of expression in DNA damage repair genes.

关 键 词：大气细颗粒物 人支气管上皮细胞 DNA损伤 单细胞凝胶电泳实验 

分 类 号：R122.7[医药卫生—环境卫生学]