Wang Ya-nan, Shuai Pei-qiang, Guo Ya-jie, Zhang Li-shi
Department of Nutrition and Food Hygiene, West China School of Public Health, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2006 Jul;37(4):558-61.
To establish TK gene mutation assay using human lymphoblastoid cell line TK6 and to study the genotoxic mechanism of Vinblastine (VBL).
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.
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.
TK6 cell line can be used to detect the genotoxicity of Vinblastine and LOH was the major mutation events in Vinblastine-induced mutants.
建立利用人淋巴母细胞系TK6的TK基因突变检测方法，并研究长春碱（VBL）的遗传毒性机制。
用不同浓度（0.625 ng/mL、1.250 ng/mL、2.500 ng/mL和5.000 ng/mL）的长春碱处理TK6细胞24小时，并进行TK基因突变检测。检测相对存活率（RS%）、相对悬浮生长率（RSG%）、tk位点的突变频率和缓慢生长突变体的百分比（SC%），并分析突变体的杂合性缺失（LOH）。
当TK6细胞分别用0.625、1.250、2.500和5.000 ng/mL的长春碱处理24小时时，观察到RS%和RSG%降低，tk位点的突变频率呈剂量依赖性增加。结果表明，约96.4%的长春碱诱导突变体为LOH。其中，半合子LOH占39.3%，纯合子LOH占57.1%。
TK6细胞系可用于检测长春碱的遗传毒性，LOH是长春碱诱导突变体中的主要突变事件。
