POLYMERASE CHAIN REACTION-BASED DELETION SCREENING OF BLEOMYCIN-INDUCED 6-THIOGUANINE-RESISTANT MUTANTS IN CHINESE-HAMSTER OVARY CELLS - THE EFFECTS OF AN INHIBITOR AND A MIMIC OF SUPEROXIDE-DISMUTASE

被引:23
作者
AN, J
HSIE, AW
机构
[1] Department of Preventive Medicine and Community Health, The University of Texas Medical Branch, Galveston
来源
MUTATION RESEARCH | 1993年 / 289卷 / 02期
关键词
CHO CELLS; REACTIVE OXYGEN SPECIES; BLEOMYCIN; SUPEROXIDE DISMUTASE; DELETION; THIOGUANINE-RESISTANT MUTANT;
D O I
10.1016/0027-5107(93)90072-N
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Bleomycin-induced 6-thioguanine-resistant mutants pretreated with or without TRIEN (triethylenetetramine), a superoxide dismutase (SOD) inhibitor, or TEMPOL (4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl), an SOD mimic, were analyzed by polymerase chain reaction (PCR)-based deletion screening in a Chinese hamster ovary (CHO) clone K1-BH4 and its derivative AS52 cells. As we proposed earlier, TRIEN would decrease and TEMPOL would increase the intracellular level of hydroxyl radical leading to a higher and lower recovery of deletion mutants. We found that the proportion of the deletion mutants induced by bleomycin at the hypoxanthine-guanine phosphoribosyltransferase (hprt) locus in K1-BH4 cells was 45.5% (25/55). The proportion of deletion HPRT- mutants induced by bleomycin pretreated with TRIEN was 31.0% (9/29) and with TEMPOL was 50.0% (14/28). The proportion of deletion mutants induced by bleomycin on the xanthine-guanine phosphoribosyltransferase (gpt) gene in AS52 cells was 61.0% (36/59). The proportion of deletion GPT- mutants induced by bleomycin pretreated with TRIEN was 56.8% (21/37) and with TEMPOL was 61.4% (27/44). The trend of the change of the proportion of bleomycin-induced deletion mutants as affected by TRIEN and by TEMPOL provides molecular evidence for the involvement of reactive oxygen species (ROS) in bleomycin mutagenesis in mammalian cells, in which deletion is a major type of induced mutation.
引用
收藏
页码:215 / 222
页数:8
相关论文
共 25 条