8-Bromo-7-Methoxychrysin Targets NF-?B And FoxM1 To Inhibit Lung Cancer Stemness Of H460 Cells Induced By Pro-inflammatory Factors

Objective:The purpose of this study was to investigate whether BrMC(8-bromo-7-methoxy-Chrysin)inhibits lung cancer stemness of H460 cells induced by inflammatory factors(TGF-? combined with TNF-?)and its potential mechanism.Methods:1.Human non-small cell lung cancer H460 cells cultured in 10% fetal bovine serum DMEM medium,establishment of lung cancer stem-like cells of NCI-H460 cell line chronically exposed to TGF-? induced by inflammatory factor TNF-?.2.Lung cancer stem-like cells induced by inflammatory factors were treated with different concentrations of BrMC.3.Combination of BrMC with NF-?B inhibitor(SN50)or FoxM1(thiostrepton,THI)inhibitor can synergistically inhibit the expression of NF-kappa B and FoxM1 and stem cell-like characteristics in NCI-H460 lung cancer stem-like cells induced by inflammatory factors4.The expression levels of NF-kappa B and FoxM1,stem cell markers(CD133,CD44)and pluripotent transcription factors(Bmi1,Oct4)were detected by Western blotting.The tumorigenicity in vitro was tested by spherical formation test and soft agar colony formation test.Results:1.Inflammatory factor TNF-alpha induces higher stem cell characteristics andincreases the expression of NF-kappa B and FoxM1 in NCI-H460 cells exposed to TGF-beta chronically.2.BrMC inhibits the characteristics of stem cells induced by inflammatory factors and the expression of NF-kappa B and FoxM1 in a dose-dependent manner.3.BrMC combined with NF-?B inhibitor(SN50)SN50 or FoxM1 inhibitor(THI,thiostrepton)can synergistically inhibit the expression of NF-kappa B and FoxM1 and the stem cell-like characteristics of NCI-H460 lung cancer stem-like cells induced by inflammatory factors.4.Overexpression of NF-kappa B or FoxM1 can attenuate the expression of NF-kappa B and FoxM1 and stem cell-like characteristics in NCI-H460 lung cancer stem-like cells by BrMR.Conclusion:1.NCI-H460 cells chronically exposed to TGF-beta induced by inflammatory factor TNF-alpha have the characteristics of tumor stem-like cells.2.BrMC inhibited lung cancer stemness,as validated by enhanced self-renewal ability,higher in vitro tumorigenicity,and increased expression of CD133,CD44,Bmi1 and Oct4 in H460 cells administered TNF-? after prolonged induction by TGF-?,in a concentration-dependent manner.