To investigate the effect of HMGB1 on inducing epithelial mesenchymal transition in human lung adenocarcinoma epithe-lial cells(A549)
Objective To investigate the effect of HMGB1 on inducing epithelial mesenchymal transition in human lung ade-nocarcinoma epithelial cells(A549).Methods A549 cells were stimulated in vitro by different concentrations of rHMGB1(0,0.5,1,2,4 μg/mL),and the relative expression of vimentin protein in each concentration group was detected by western blot assay,and the optimal in vitro action concentration of rHMGB1 was derived as 2 μg/mL.Then the experiments were divided into three groups,HMGB1 group,TGF-β group(as positive control),control group(as negative control).2 μg/mL rHMGB1,2 μg/mL TGF-β and RPMI1640 medium only were used to stimulate A549 cells in vitro,separately.And after incubation at 37℃and 5%CO2 for 48 h or 96 h,MTT assay was performed to detect the proliferation rate of each group,transwell assay was performed to detect the migra-tion ability of each group,and the relative expression of E-cadherin,vimentin and snail1 protein in each group was detected by western blot assay.Results The proliferation rate,the number of cell migration,the relative expression of intracellular snail1 and vimentin proteins were significantly increased(P<0.0001 or P<0.01)and the relative expression of E-cadherin protein was signifi-cantly decreased(P<0.0001)in A549 cells of the HMGB1group and the TGF-β group compared with the control group.Conclu-sion HMGB1 has the effect of inducing epithelial mesenchymal transformation of human lung adenocarcinoma epithelial cells A549 in vitro.
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