【作者】 胡大林； 林忠宁； 刘移民； 袁建辉； 纪卫东； 方道奎； 涂晓志； 庄志雄；

【Author】 HU Da-lin,LIN Zhong-ning,LIUYi-ming,et al (1.Department of preventive medicine,Medical college,Foshan University,Foshan,China.528000; 2.Department of preventive medicine,School of Public Health,Sun Yat-san University,Guangzhou,China, 510080;3.Shenzhen Center for Disease Control and Prevention,518020;4.Guangzhou Occupational Disease Hospital,Guangzhou,China,510420)

【机构】 佛山科学技术学院医学院； 中山大学公共卫生学院预防医学系； 广州市职业病防治院； 深圳市疾病预防控制中心；

【摘要】 目的探讨氢醌(HQ)对人DNA聚合酶β(polβ)基因低表达细胞的毒作用。方法以人支气管上皮细胞(HBE)为靶细胞,运用载体介导的RNA干扰技术抑制polβ基因的表达,构建polβ低表达细胞株(HBE-β~-);以不同浓度的HQ(0、10、20、40、80、120μmol/L)对HBE-β~-细胞、转染了空载体的细胞(HBE-pEGFP-C1)及正常对照组细胞(HBE)进行染毒;光学显微镜下观察染毒细胞的形态学改变;以MTT法测定HQ的细胞毒性;以彗星实验(Comet assay)测定DNA的损伤程度。结果光镜下可见染毒细胞出现不同程度的收缩和漂浮现象;经不同浓度HQ染毒的HBE-β~-细胞OD值均显著低于两种对照组细胞(P<0.05),两对照组细胞OD值差异不明显(P>0.05),三组细胞的OD值与HQ浓度之间呈现一定的剂量反应关系;Comet assay以彗星全长(Total length)均数±SD来评价,3组细胞DNA均有损伤,且与HQ浓度之间存在一定的剂量反应关系;HBE-β~-细胞的DNA彗星全长均数与两对照组有显著差异(P<0.05),而两对照组彗星全长均数之间无明显差异(P>0.05)。结论在体外实验条件下,HQ具有明显的细胞毒性,DNA polβ低表达可增强HQ对DNA分子的损伤程度。更多还原

【Abstract】 Objective To explore the toxicity of hydroquinone(HQ) to human bronchial epithelial cells(HBE) when DNA polymerase beta(po1β)was knocked down by RNA interference(RNAi).Methods Technology of vector-mediated RNAi was used to knock-down po1βin HBE,then cell line of‘HBE-β’ was obtained.The control groups included HBE and the cell line of‘HBE-pEGFP-C1’ of which transfection of the carrier of‘pEGFP-C1’’was success.Various concentration of HQ(0、10、20、40、80、120μmol/L)was separately added to the up related three cell groups.Morphological characteristics of each group were analyzed under light microscope.MTT assay was used to analyze the cytotoxicity of hydroquinone.The degree of DNA damage was analyzed by Comet assay.Results Cells in three groups shrinked and floated when HQ was added into the culture medium.The OD value of experiment group was transparently lower than that of the two control groups(P<0.05).While there was no obviously differences between the two control groups(P>0.05).Dose response relation was found between OD value and HQ in three groups.Mean±SD of total length of DNA comet acted as index,the obviously DNA damage was found in three groups,and there was dose response relation between DNA damage and HQ,at the same time,there was obviously difference in the degree of DNA damage among experiment group and two control groups(P<0.05),while there no transparent difference between the two control groups(P>0.05).Conclusion Obviously cytotoxicity of HQ was found under the experiment condition in vitro,the enhancement of cytotoxicity of HQ to DNA molecule was found when DNA poipgene was knocked-down.更多还原