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Published:
Jan 12, 2017Keywords:
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Main Article Content
Effect of dose and dosing rate on the mutagenesis of nitric oxide in supF shuttle vector
Abstract
Purpose: To determine how the dose and rate of NO• treatment affects mutagenic responses.
Methods: Shuttle vector pSP189 was used to determine the genotoxicity resulting from in vitro exposure to NO• using three delivery methods (reactor and Transwell co-culture systems, and NO• donor sodium nitroprusside), followed by plasmid replication in bacteria MBL50 and human AD293 cells.
Results: When exposed to preformed 100% NO• for 3 h or 1% NO• for 35 h using a reactor system, a cumulative dose of 1260 μM × min reduced AD293 cell viability by 46 and 18% and increased mutation frequencies (MFs) 1.9- and 5.3-fold higher than argon control, respectively. Roughly 5-fold increase in MF of the supF gene of AD293 cells co-cultivated with macrophages stimulated with IFN-γ/LPS was also observed. When AD293 cells were treated by SNP, DNA strand breaks were induced and MFs were increased in a dose-dependent manner.
Conclusion: These results provide important clues to how dose and dosing rate of introducing NO• may contribute to potential genotoxicity resulting from NO• formation in vivo.
Keywords: AD293 cells, Delivery method, Genotoxicity, Nitric oxide, supF Gene of pSP189 shuttle vector
