体内Pig-a基因突变试验高通量方法联合验证研究

doi:10.3969/j.issn.1004-616x.2013.05.016

Abstract

Abstract:

OBJECTIVE: To evaluate the reproducibility and transferability of the erythrocyte(RBC) based Pig-a mutation assay and explore the potential of combining this assay and micronucleus(MN) analyses into one study. METHODS：20 rats were randomly assigned to be treated with PBS and different doses of (10，20 and 40 mg/kg) N-ethyl-N-nitrosourea (ENU) for 3 consecutive days by oral gavage. Jugular blood samples were collected on days -1 (the day before administration)，14，and 30 and evaluated for Pig-a mutantion frequencies in whole peripheral blood RBCs and reticulocytes (RETs). Day 4 samples were scored for micronucleated reticulocyte frequencies. RESULTS： While ENU-induced Pig-a mutation frequencies were consistent with previously reported results，analysis rate and number of cells evaluated were dramatically increased. ENU also induced significant increases in MN-RET frequencies. CONCLUSION： The in vivo Pig-a mutation high-throughput assay was established successfully in rat. Results with ENU indicated that the new Pig-a scoring methodology was reproducible Pig-a mutation and MN analyses could be readily combined into one study.

Key words: N-ethyl-N-nitrosourea, Pig-a mutation, immunomagnetic separation, micronucleus, flow cytometry

ZHANG Ming，ZHOU Chang-hui，CHANG Yan，WANG Zheng，TU Hong-gang，LI Jie. Interlaboratory transferability study of the Pig-a mutation assay with immunomagnetic enrichment[J]. Carcinogenesis, Teratogenesis & Mutagenesis, doi: 10.3969/j.issn.1004-616x.2013.05.016.

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Interlaboratory transferability study of the Pig-a mutation assay with immunomagnetic enrichment

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