中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (15): 2831-2834.doi: 10.3969/j.issn.1673-8225.2012.15.038

• 组织构建基础实验 basic experiments in tissue construction • 上一篇    下一篇

互隔交链孢酚对NIH/3T3细胞中DNA聚合酶β的致突变作用*★

朱  涵，樊红琨，章  茜   

  1. 郑州大学医学院生理学教研室，河南省郑州市 450052
  • 收稿日期:2011-12-02 修回日期:2012-01-10 出版日期:2012-04-08 发布日期:2012-04-08
  • 通讯作者: 章茜，博士，教授，博士生导师，郑州大学医学院生理学教研室，河南省郑州市 450052 qianzhang@zzu.edu.cn
  • 作者简介:朱涵★，女，1977年生，河南省镇平县人，汉族，硕士，2004年郑州大学医学院毕业，讲师，主要从事肿瘤发病机理研究工作。 zhuhan@zzu.edu.cn
  • 基金资助:

    教育部科学技术研究重点项目(204084)，项目名称：DNA聚合酶β在互隔交链孢霉诱发癌变中的表达及修复作用的研究。

Mutation effect of alternariol on DNA polymerase beta in NIH/3T3 cell line

Zhu Han, Fan Hong-kun, Zhang Qian   

  1. Department of Physiology, Medical College of Zhengzhou University, Zhengzhou  450052, Henan Province, China
  • Received:2011-12-02 Revised:2012-01-10 Online:2012-04-08 Published:2012-04-08
  • Contact: author: Zhang Qian, Doctor, Professor, Doctoral supervisor, Department of Physiology, Medical College of Zhengzhou University, Zhengzhou 450052, Henan Province, China qianzhang@zzu.edu.cn
  • About author:Zhu Han★, Master, Lecturer, Department of Physiology, Medical College of Zhengzhou University, Zhengzhou 450052, Henan Province, China zhuhan@zzu.edu.cn
  • Supported by:

    Ministry of Education Key Project of Science and Technology, No.204084*

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摘要：

背景：近年来研究发现食管癌细胞中存在DNA聚合酶β的突变。
目的：观察互隔交链孢酚对NIH/3T3细胞中DNA聚合酶β基因序列的影响，研究互隔交链孢酚致细胞变异的机制。
方法：用不同浓度(2，4，6，8 μmol/L)的互隔交链孢酚作用于NIH/3T3细胞，并设置对照组。用RT-PCR法从细胞总RNA中扩增出DNA聚合酶β基因cDNA序列，应用T-A克隆技术，克隆至pGEM-T载体后进行测序，分析其序列变化。
结果与结论：2 μmol/L互隔交链孢酚处理后的细胞内DNA聚合酶β基因序列无任何变化。而4 μmol/L组中DNA聚合酶β基因有1个位点发生突变，8 μmol/L组和16 μmol/L组中各有2个位点发生突变，此3组的突变存在相同的位点。结果证实，互隔交链孢酚可导致NIH/3T3细胞中的DNA聚合酶β发生点突变，互隔交链孢酚浓度越高，点突变的数量越多，且存在突变活跃位点。  
                       

关键词: 互隔交链孢酚, DNA聚合酶&beta, NIH/3T3细胞, 克隆, 突变, 组织工程

Abstract:

BACKGROUND: The mutation of DNA polymerase β (DNA pol β) has been found in esophageal carcinoma cells.
OBJECTIVE: To analyze the effect of alternariol on the sequence of DNA pol β in NIH/3T3 cell line and to study the mutation mechanism of alternariol
METHODS: The NIH/3T3 cell line was injected with different concentrations of alternariol (2, 4, 6, 8 μmol/L) and the control group was set. DNA pol β gene was amplified from the total RNA of the cells using reverse transcription-PCR, then cloned with pGEM-T vector and sequenced with T-A cloning technique, and the sequence was analyzed.
RESULTS AND CONCLUSION: The sequence of DNA pol β was not changed in 2 μmol/L alternariol group, and there was one mutation point in 4 μmol/L alternariol group and two mutation points in 8 and 16 μmol/L alternariol group. There was a same mutation point in these three groups. Alternariol can induce point mutation of DNA pol β gene in NIH/3T3 cell line. Higher concentration of alternariol, more points of DNA pol β gene mutation. The active mutation point exists.
 

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