摘要
目的 :建立WTK1细胞tk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tk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tk位点突变率增加 ,MMS的处理剂量达到10mg·L-1 时 ,tk位点突变率为985.2/106 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tk位点诱发了两种不同表型的突变集落 ,即正常生长突变体(tk_NGmutant)和慢生长突变体(tk_SGmutant) ,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tk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tk基因突变试验 。
Purpose: To develop a method of t k gene mutation assay in WTK1 cell for testing gene mutation induced by c hemical mutagens. Methods: t k locus mutation frequency and p53 gene protein expression level were detected after WTK1 lymphob lastoid cells were treated with methyl methanesulfonate (MMS). R esults: 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 mutati on colonies, namely t k_NG and t k_SG mutant colonies, but main ly 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 lymphob lastoid cell, and WTK1 cell can be used in mutagenesis test at t k locus.
出处
《癌变．畸变．突变》
CAS
CSCD
2003年第2期116-119,共4页
Carcinogenesis,Teratogenesis & Mutagenesis
基金
国家自然科学基金资助项目(No.39770653)
关键词
WTK1细胞
TK基因
突变
甲基磺酸甲酯
WTK1 cell
thymidine kinase(t k) gene
mutation
m ethyl methanesulfonate(MMS)
