摘要
目的建立用人类淋巴母细胞TK 6检测纺锤体毒物——长春花碱的TK基因突变试验方法,同时探讨长春花碱的遗传毒性分子机理。方法使用长春花碱0.625 ng/mL、1.250 ng/mL、2.500 ng/mL和5.000 ng/mL对TK 6细胞染毒24 h,进行TK基因突变试验。检测细胞相对存活率(RS%)、相对悬浮增长率(RTG%)、tk位点突变频率(M F)和缓慢生长集落的百分比(SC%),同时对突变集落tk位点杂合性丢失(LOH)进行分析。结果随着长春花碱浓度的增加,TK 6细胞的相对存活率和相对悬浮增长率均降低,而tk位点突变频率逐渐增加,且有剂量反应关系。tk位点杂合性分析表明长春花碱诱发的突变集落中有96.4%为LOH丢失,其中半合子LOH为39.3%,纯合子LOH为57.1%。结论长春花碱可诱导TK 6细胞tk基因突变,主要以LOH为主。
Objective To establish TK gene mutation assay using human lymphoblastoid cell line TK6 and to study the genotoxic mechanism of Vinblastine（VBL）. Methods TK6 cells were treated with Vinblastine at different concentrations （0.625 ng/mL, 1.250 ng/mL, 2.500 ng/mL and 5.000 ng/mL）for 24 h and TK gene mutation assay were experimented. Relative survival （RS%）, relative suspension growth （RSG%）, mutation frequency at tk locus and percentages of slow growth mutant （SC%） were detected and loss of heterozygosity （LOH） of mutants were analyzed. Results A decreased RS% and RSG% and an increased mutation frequency at tk locus were observed in a dose-dependent manner when the TK6 cells were treated with 0.625, 1.250, 2.500 and 5.000 ng/mL of Vinblastine respectively for 24 h. The result demenstrated that about 96.4% of Vinblastine-induced mutants were LOH. Among them, 39.3% were hemi-LOH and 57.1% were homo-LOH respectively. Conclusion TK6 cell line can be used to detect the genotoxicity of Vinblastine and LOH was the major mutation events in Vinblastine-induced mutants.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2006年第4期558-561,共4页
Journal of Sichuan University(Medical Sciences)
基金
国家自然科学基金(批准号30471473)
博士点基金(批准号20020610036)
