癌变·畸变·突变 ›› 2003, Vol. 15 ›› Issue (2): 116-119.doi: 10.3969/j.issn.1004-616x.2003.02.018

• 论文 • 上一篇    下一篇

甲基磺酸甲酯诱发WTK1细胞t k位点突变试验方法的建立

张建清1, 张立实2, 王瑞淑2   

  1. 1. 深圳市疾病预防控制中心, 广东 深圳 518020; 2. 华西医科大学公共卫生学院, 四川 成都 610041
  • 收稿日期:2002-06-17 修回日期:2002-10-11 出版日期:2003-04-30 发布日期:2003-04-30
  • 通讯作者: 张建清

INDUCTION MUTATION OF MMS AT t k LOCUS IN WTK1 HUMAN DIPLOID LYMPHOBLASTOID CELL

ZHANG Jian-qing 1, ZHANG Li-shi 2, WANG Rui-shu 2   

  1. 1. Center for Disease Control, Shenzhen 518020 , China; 2. School of Public Health, West China University of Medical Science, Chengdu 610041 , China
  • Received:2002-06-17 Revised:2002-10-11 Online:2003-04-30 Published:2003-04-30
  • Contact: ZHANG Jian-qing

摘要: 目的: 建立WTK1细胞t k基因突变试验方法,为毒理学评价和机制的研究提供实验依据。 方法: 用标准诱变剂甲基磺酸甲酯(MMS)处理WTK1细胞,检测t k位点突变率和细胞p53基因蛋白表达水平的改变。 结果: MMS可诱导WTK1细胞t k位点突变率增加,MMS的处理剂量达到10 mg•L-1时,t k位点突变率为985.2 / 10 6个细胞,并有剂量反应关系。诱发突变率约是自发突变率3~10倍。 在t k位点诱发了两种不同表型的突变集落,即正常生长突变体(t k-NG mutant)和慢生长突变体(t k-SG mutant),但以慢生长突变体为主。 MMS处理后,WTK1细胞P53蛋白的表达水平增高。 结论: MMS可以诱发WTK1细胞t k位点的突变率增加,MMS处理细胞后,P53蛋白的表达水平增高,说明WTK1细胞的P53蛋白的功能尚未丧失。该方法的建立,为进一步开展t k基因突变试验,评价环境化学物的诱变性奠定了基础。

关键词: WTK1细胞, t k基因, 突变, 甲基磺酸甲酯

Abstract: Purpose: To develop a method of t k gene mutation assay in WTK1 cell for testing gene mutation induced by chemical mutagens. Methods: t k locus mutation frequency and p53 gene protein expression level were detected after WTK1 lymphoblastoid cells were treated with methyl methanesulfonate (MMS). Results: MMS induced t k locus mutation with mutation frequency 3~10 times higher than that of spontaneous mutation frequency of WTK1 cell. Induction mutation frequency depended on dose level. t k gene mutation frequency (MF) was 985.2 / 10 6 cells at 10 mg•L-1. There were two different phenotypes of mutation colonies, namely t k-NG and t k-SG mutant colonies, but mainly t k-SG mutant colonies. The level of P53 protein expression increased after treatment of WTK1 cell with MMS. Conclusion: MMS inducted t k locus mutation in WTK1 human diploid lymphoblastoid cell, and WTK1 cell can be used in mutagenesis test at t k locus.

Key words: WTK1 cell, thymidine kinase(t k) gene, mutation, methyl methanesulfonate(MMS)