南方医科大学学报

南方医科大学学报 ›› 2006, Vol. 26 ›› Issue (08): 1092-1095.

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低剂量的氢醌诱导细胞损伤耐受差异表达基因的研究

卫秦芝； 庄志雄；   

  1. 南方医科大学公共卫生与热带医学学院毒理学系； 深圳市疾病预防控制中心 广东广州510515； 广东深圳518020；
  • 出版日期:2006-08-20 发布日期:2006-08-20
  • 基金资助:
    国家科技攻关973资助项目(2002CB512904)~~

Differentially express genes in human embryonic lung fibroblasts with damage tolerance induced by low-dose hydroquinone

WEI Qin-zhi1, ZHUANG Zhi-xiong2 1Department of Health Toxicology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, China; 2Disease Control and Prevention Center of Shenzhen, Shenzhen 518020, China   

  1. 南方医科大学公共卫生与热带医学学院毒理学系； 深圳市疾病预防控制中心 广东广州510515； 广东深圳518020；
  • Online:2006-08-20 Published:2006-08-20

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摘要： 目的探讨低剂量氢醌(HQ)诱导人胚肺成纤维细胞(HLF)损伤耐受差异表达的基因。方法参照本实验室用MTT法所得的HQ对HLF毒性的剂量效应关系,选择100pmol/L的HQ为低剂量,100μmol/L的HQ为高剂量,按以下方式对细胞进行染毒:空白对照、低剂量、高剂量、损伤耐受组(先用低剂量预刺激24h,再用高剂量刺激6h,然后用荧光差异显示PCR寻找不同方式刺激差异表达的基因,并对差异表达基因进行克隆、鉴定同源性比较。结果按方法所示的方式处理细胞,处理完毕后提取各组细胞总RNA,进行荧光差异显示PCR,找到33个差异条带。对其中的8个差异条带进行克隆鉴定同源性比较,发现其中1个已知基因,7个未知基因。结论用荧光差异显示PCR方法寻找HQ诱导HLF损伤耐受过程差异表达的基因,通过鉴定基因,为进一步研究低剂量HQ诱导HLF损伤耐受的机制提供科学依据。 

Abstract: Objective To observe the differentially expressed genes in human embryonic lung fibroblasts (HELF) induced by small-dose hydroquinone (HQ) using fluorescence differential display-PCR (DD-PCR). Methods According to the dose-effect relation of HQ toxicity we established previously, HQ dose that did not induce observed cell damage or proliferation arrest was defined as low dose (100 pmol/L), and that causing obvious cell damage as the high dose (100 μmmol/L). The cells were then treated with low or high dose of HQ, or exposed to high-dose HQ following pretreatment with low-dose HQ for some time, respectively. Fluorescence DD-PCR was performed and 33 differentially expressed genes were identified in the cells with different treatments, and 8 of the identified genes were amplified, cloned, sequenced and blasted. Results Seven of the 8 amplified genes were unknown genes, and the left one was identified as a known gene highly homologous to that encoding Homo sapiens Rap1 interacting factor 1 (RIF1). Conclusion Low-dose HQ can induce damage tolerance in HELF, and identification of the differentially expressed genes may provide valuable sight into the mechanism of HQ-induced damage tolerance. 

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