β-Naphthoflavone Can Inhibit Luciferase Activity

Objective:To investigate the effect of β-Naphthoflavone on the firefly luciferase activity.Methods:A549,HepG2,HeLa,MCF-7,Bel-7402,16HBE cells were transfected with GCLC5'-upstream regulatory sequence driven PGL3-enhancer-Luciferase reporter vector(PL45)and treated with β-NF.The dual-luciferase reporter assay system was used to analyze the effect of β-Naphthoflavone on the expression of GCLC gene.Western blot was used to detect the change of protein level.A549 and HepG2 cells were transfected with the eukaryotic expression vector pRC/CMV2-luc+ and treated with β-NF.The dual-luciferase reporter assay system was used to analyze the effect of β-Naphthoflavone on the firefly luciferase activity.A549 and HepG2 cells were transfected with the PL45 vector,cells were lysed,then treated with β-NF for 25 min to analyze the effect of β-NF on the firefly luciferase activity.Results:In all the cells,the relative luciferase activity of β-NF treatment group was significantly lower compared with that in DMSO control group(P0.01).The result of western bot showed that the expression of GCLC were higher inβ-NF treatment group than that in DMSO control group.In A549 and HepG2 cells,after transfected with pRC/CMV2-luc+ vector,the relative luciferase activity of β-NF treatment group was lower than that in DMSO control group(P0.01).A549 and HepG2 cells were transfected with PL45 vector,then lysed,and the relative luciferase activity in β-NF treatment group was lower than that in DMSO control group(P0.01).Conclusion:β-Naphthoflavone directly inhibits Firefly Luciferase activity.