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奉水东, 李华文, 王小利, 凌宏艳, 徐顺清. 氮芥所致DNA损伤分子标志物探讨[J]. 中国公共卫生, 2005, 21(2): 149-151.
引用本文: 奉水东, 李华文, 王小利, 凌宏艳, 徐顺清. 氮芥所致DNA损伤分子标志物探讨[J]. 中国公共卫生, 2005, 21(2): 149-151.
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FENG Shuidong, LI Huawen, WANG Xiaoli, . Study on molecular biomarker of DNA lesion induced by nitrogen mustard[J]. Chinese Journal of Public Health, 2005, 21(2): 149-151.
Citation: FENG Shuidong, LI Huawen, WANG Xiaoli, . Study on molecular biomarker of DNA lesion induced by nitrogen mustard[J]. Chinese Journal of Public Health, 2005, 21(2): 149-151.
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氮芥所致DNA损伤分子标志物探讨

Study on molecular biomarker of DNA lesion induced by nitrogen mustard

    摘要
  • 摘要:
      目的   利用烷化剂的毒理机制, 评价生长抑制和DNA损伤(Growth arrest and DNA damage, Gadd)基因的表达作为DNA损伤的分子标志物的可行性。
      方法   通过烷化剂类药物氮芥诱导人肝瘤HepG2细胞, 运用RT-PCR的方法检测Gadd基因的表达水平, 同时佐以MTT试验、单细胞凝胶电泳试验来分析氮芥对细胞的毒性作用和DNA损伤程度。
      结果   氮芥能诱导Gadd基因不同程度的表达, 但随着浓度和时间的增加有由表达增强逐渐转为抑制的趋势; 氮芥对人肝瘤HepG2细胞能产生明显的DNA损伤作用, 有浓度依赖关系(rs=0.591, P < 0.01);氮芥对细胞有毒性作用, 不同浓度的氮芥(0.5~40μg/L)使细胞存活率从97%下降到52%。
      结论   Gadd基因的表达可以作为判断DNA损伤的比较敏感的分子标志物。

     

    Abstract:
      Objective   To assess the feasibility of expression level of Growth arrest and DNA damage (Gadd) genes function as molecular biomar ker of DNA lesion induced by nitrogen mustard, according to the toxicity mechanism of alkylating agents.
      Methods   The human liver tumor HepG2 cells were first treated with nitrogen mustard, the mRNA level of Gadd45, Gadd53, Gadd34 genes were determined by RT-PCR.Meanwhile the single cell gel electrophoresis (comet assay) was applied to determine the DNA damage induced by varying doses of nitrogen mustard and the 3-(4, 5-methylthiozol-2-yl)-2, 5-diphenylte-trazolium bromide (MTT) metabolic viability assay was performed to assess the cytotoxicity of nitrogen mustard.
      Results   Treated with 1 ng/μl nitrogen mustard for different time, expression of Gadd genes increased from 3h to 24h and then decreased gradually.Treated with 1 ng/μl and 5 ng/μl nitrogen mustard respectively for 24 h, expression of Gadd genes had a tendency with from rising to inhibiting.With the concentration of nitrogen mustard increased, damaged cells increased (rs=0.591, P < 0.01).Treatment with nitrogen mustard of varying concentration (0.5 ng-40 ng/μl) decreased cell viability from 97% to 52% respectively.
      Conclusion   It is possible of expression level of Gadd genes to function as molecular biomarker of DNA damage induced by nitrogen mustard.

     

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