细胞外组蛋白调控TLR4/NF-κB途径对膀胱癌细胞上皮间质转化和侵袭的影响

Effect of Extracellular Histone on Epithelial Mesenchymal Transformation and Invasion of Bladder Cancer Cells by Regulation of TLR4/NF-κB Pathway

目的研究细胞外组蛋白调控Toll样受体4(Toll like receptor 4,TLR4)/核因子-κB(nuclear factor-κB,NF-κB)途径对膀胱癌细胞上皮间质转化(epithelial mesenchymal transformation,EMT)和侵袭的影响。方法培养膀胱癌细胞株T24并分为对照组、不同浓度组蛋白H3(50、100、200μg/mL)组、溶剂对照(体积分数0.1%DMSO)组、组蛋白H3(200μg/mL)+溶剂对照(体积分数0.1%DMSO)组、组蛋白H3(200μg/mL)+NF-κB抑制剂Bay11-7082(10μmol/L)组。分组给药24 h后检测细胞侵袭数目及细胞中E-钙粘蛋白(E-cadherin)、N-钙粘蛋白(N-cadherin)、波形蛋白(Vimentin)、TLR4、磷酸化P65 NF-κB(p-P65 NF-κB)的蛋白表达水平。结果不同浓度组蛋白H3组的细胞侵袭数目及N-cadherin、Vimentin、TLR4、p-P65 NF-κB的表达水平高于对照组,E-cadherin的蛋白表达水平低于对照组(P<0.05)且组蛋白H3浓度越高,细胞侵袭数目及蛋白表达水平的变化越显著;组蛋白H3+Bay11-7082组的细胞侵袭数目及N-cadherin、Vimentin、p-P65 NF-κB的表达水平低于组蛋白H3+溶剂对照组,E-cadherin的蛋白表达水平高于组蛋白H3+溶剂对照组(P<0.05)。结论细胞外组蛋白促进膀胱癌细胞EMT及侵袭,这一促进作用与激活TLR4/NF-κB途径有关。

Objective To investigate the effect of extracellular histone on epithelial mesenchymal transformation(EMT)and invasion of bladder cancer cells by regulating toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)pathway.Methods T24 bladder cancer cells were cultured and divided into 7 groups:control group,histone H3 groups with different concentrations(50,100,200μg/mL),solvent control(0.1%DMSO)group,histone H3(200μg/mL)+solvent control(0.1%DMSO)group,and histone H3(200μg/mL)+NF-κB inhibitor Bay11-7082(10μmol/L)group.The number of invasion cells and the protein expression levels of E-cadherin,N-cadherin,Vimentin,TLR4 and phosphorylated P65 NF-κB(p-P65 NF-κB)were detected 24 h after administration.Results The number of invasion cells and the expression levels of N-cadherin,Vimentin,TLR4 and p-P65 NF-κB in the histone H3 groups were higher than those in the control group,and the protein expression level of E-cadherin was lower than that in the control group(P<0.05),the number of invasion cells and the expression levels of those proteins were changed with the concentration of histone H3 treatment in a dose-dependent manner.The number of cell invasion and the protein expression levels of N-cadherin,Vimentin and p-P65 NF-κB in the histone H3+Bay11-7082 group were lower than those in the histone H3+solvent control group,and the expression level of E-cadherin was higher than that in the histone H3+solvent control group(P<0.05).Conclusion Extracellular histone promotes EMT and invasion of bladder cancer cells,which is related to the activation of TLR4/NF-κB pathway.