Objective:Rapid detection of DNA damage caused by trace fine particulate matter(PM2.5)using recombinant human lung epithelial cells(BEAS-2B).Methods:Different concentrations of PM2.5 were prepared and applied to recombinant lung epithelial cells.The survival rate of recombinant human lung epithelial cells was detected by MTS(Internal salt method,3-(4,5-dimethylthiazole-2)-5-(3-carboxymethylphenyl)-2-(4-sulfophenyl)-2H-tetrazole internal salt method),DNA damage was detected by comet assay,GADD153 mRNA gene expression was detected by RT-PCR,and DNA damage was detected by luciferase activity under different concentrations.Results:Compared with negative control,25,125 μg/mL PM2.5 significantly decreased the activity of recombinant cells and significantly increased the DNA damage(comet assay).Compared with negative control,5,25,125 μg/mL PM2.5 significantly increased the endogenous GADD153 mRNA gene expression and luciferase activity of recombinant cells.GADD153 mRNA gene expression and luciferase activity had a good correlation(r2=0.992,P＜0.01).Conclusion:Compared with the traditional methods for DNA damage in cells,recombinant human lung epithelial cells based on GADD153-LUC can rapidly detect DNA damage caused by lower dose PM2.5.
Fine particulate matterLung epithelial cellsDNA damageGadd1 53-luc gene
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