氮芥染毒Neuro-2a细胞生成DNA加合物的时效及量效关系研究OA

Objective To identify the biomarkers of effect in DNA damage induced by nitrogen mustard treatment.Methods Neuro-2a cells were subjected to in vitro treatment with nitrogen mustards(HN1,HN2 or HN3).The types and levels of HN-DNA adducts were quantitatively analyzed using the multiple reaction monitoring(MRM)mode coupled with ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS).Results Good linearity was observed in the nine HN-DNA adduct standards within the range of 0.02 to 100 ng/mL,with coefficients of determination above 0.99.The limits of detection(LOD)and limits of quantitation(LOQ)ranged from 0.01 to 0.02 ng/mL and from 0.02 to 0.05 ng/mL,respectively.The relative standard deviations(RSD)for intra-assay and inter-assay precision were less than 15％.Such adducts as HN-N3-adenine(AN3),HN-N7-guanine(GN7),and HN-Bis-GN7 were detected in cells treated with the three nitrogen mustards.Adduct levels increased immediately after treatment and decreased 8 to 12 h later,showing a positive correlation with the dose of treatment.Conclusion By dynamically monitoring the temporal and quantitative changes of DNA adducts in Neuro-2a cells,this study confirms their potential as effect biomarkers in nitrogen mustard treatment while providing data for investigating the repair processes and mechanisms of DNA damage induced by nitrogen mustards.