Analysis of spontaneous, gamma ray-and ethylnitrosourea-induced hprt mutants in HL-60 cells with multiplex PCR
Analysis of spontaneous, gamma ray-and ethylnitrosourea-induced hprt mutants in HL-60 cells with multiplex PCR作者机构：Department of Healath Toxicology Preventive Medical College Third Military Medical University Chongqing 400038 China.
出 版 物：《World Journal of Gastroenterology》 (世界胃肠病学杂志（英文版）)
年 卷 期：2003年第9卷第3期
页 面：578-583页
核心收录：
学科分类：1002[医学-临床医学] 100214[医学-肿瘤学] 10[医学]
基 金：the National Natural Science Foundation of China No.39970650
主 题：人次黄嘌呤鸟嘌呤转磷酸糖基酶 HL-60细胞 γ射线 多重PCR 基因突变 分子生物学
摘 要：AIM: To explore the molecular spectra and mechanism of human hypoxanthine guanine phosphoribosyl transferase (hprt) gene mutation induced by ethyluitrosourea (ENU) and 60Co γ-***: Independent human promyelocytic leukemia cells (HL-60) mutants at the hprt locus were isolated from untreated, ethyluitrosourea (ENU) and 60Co γ-ray-exposed cells, respectively, and verified by two-way screening. The genetic changes underlying the mutation were determined by multiplex polymerase chain reaction (PCR) amplification and electrophoresis ***: With dosage increased, survival rate of plated cell reduced (in the group with dosage of ENU with 100-200μg/ml, P0.01; in the group with dosage of 60Co γ-ray with 2-4 Gy, P0.05) and mutational frequency increased (in the group of ENU 12.5-200.0 μg/ml, P0.05; in the group of 60Co γ-ray with 1-4 Gy, P0.05) significantly. In the 13spontaneous mutants analyzed, 92.3 % of mutant clones did not show any change in number or size of exon, a single exon was lost in 7.7 %, and no evidence indicated total gene deletion occurred in nine hprt exons. However, deletions were found in 79.7 % of ENU-induced mutations (62.5-89.4 %,P0.01) and in 61.7 % of gamma-ray-induced mutations (28.6-76.5 %, P0.01). There were deletion mutations in all 9 exons of hprt gene and the most of induced mutations were chain deletion with multiplex exons (97.9 % in gammaray-induced mutants, 88.1% in ENU-induced mutants).CONCLUSION: The spectra of spontaneous mutations differs completely from that induced by EUN or 60Co γ-*** both ENU and γ-ray can cause destruction of genetic structure, mechanism of mutagenesis between them may be different.
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