Objective: Different mechanisms of action compounds were studied based on Pig-a gene mutation test in L5178Y cells in vitro. Methods: A total of 10 compounds,including aristolochic acid I（AAI）, N-nitrosodiethylamine（NDEA）,methyl methanesulfonate（MMS）,cyclophosphamide（CP）, colchicine（COL）, mitomycin C（MMC）, saccharin, ethephon, benzopyrene[B（a）P] and 4-nitroquinoline-N-oxide（4-NQO） respectively at different concentration ranges were treated with L5178Y cells for a period and subsequently expressed 8 days．The cells were collected and incubated with APC-anti-CD45 and PE-anti-CD90.2,and cells with a phenotype as CD90-/CD45+ were recognised by flow cytometry as mutant in a total of million cells for calculating the mutation rates. Results: Mutagens AAI, NDEA, MMS, MMC, B（a）P and 4-NQO induced significant increases on the Pig-a gene mutation rates in L5178Y cells;while the results for CP as mutagen only after metabolism,euploid clastogen COL,non-genotoxic carcinogens ethephon and saccharin were negative. Conclusion: The in vitro Pig-a gene mutation assay based on L5178Y cells can effectively detect mutagens in both absence and presence the metabolic activation,and has showed high specificity and reliability．It is indispensable in future risk evaluation and underlying mechanism research of the drug induced gene mutation.
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