Influnce Of The Interaction Between The Mon-onuclear Cells And Fibroblasts With H460 On The Expression Of MT1-MMP In Co-cultures And The Inhibition Of NS398 To MMPs

Objective:To instigative the influence of interactions between mononuclear cells and fibroblasts and lung cancer cells (H460) on MT1-MMP,MMP-2 expression in monolayer culture model and its intervention with NS398Methods:Cells cultured in monolayer culture model were divided into H460 cells group, mononuclear cells(fibroblasts) group and co-culture group of H460 cells, mononuclear cells(fibroblasts). Immunofluorescence assay and western blot were used to analyse the expression of MT1-MMP protein levels in every group. Enzyme-linked immunosorbentassay was used to show the actions of mononuclear cells on the activation of MMP-2 in every group. Cells cultured in monolayer culture model were divided into experimental group and control group. And the experimental group was also divided into two subgroups according to the concentration of the NS398(50,100,200μmol/L). The H460 cell growth inhibition rate was detected by MTT assay. Immunofluorescence assay and western blot were used to analyse the expression of RECK and MT1-MMP protein levels in H460 cells.Results:The mean fluorescence intensity of H460 cells group, mononuclear cells group and co-culture group were 0.31±0.02,0.04±0.01,0.43±0.04. As compared with mononuclear cells group, the mean fluorescence intensity of co-culture group was higher(P<0.05). The IOD of H460 cells group, mononuclear cells group and co-culture group were 0.76±0.06,0.06±0.02,1.19±0.09. As compared with mononuclear cells group, the IOD of co-culture group was higher(P<0.05). The concentration of H460 cells group, mononuclear cells group and co-culture group were 8.20±0.49μg/L,0.13±0.01μg/L.11.32±0.75μg/L. As compared with mononuclear cells group, the concentration of co-culture group was higher(P<0.05).The IOD of H460 cells group, fibroblasts group and co-culture group were: 0.74±0.01,0.71±0.02,1.33±0.06. As compared with fibroblasts group, the IOD of co-culture group was higher(P<0.05). The concentration of H460 cells group, fibroblasts group and co-culture group were 7.92±0.22μg/L,8.94±0.27μg/L, 12.62±0.72μg/L. As compared with fibroblasts group, the concentration of co-culture group was higher(P<0.05). The growth inhibition rate of control group and experimental groups (50,100,200μmol/L) were 0,0.35±0.04,0.44±0.04,0.55±0.05. As compared with control group, NS398 caused a dose-dependent increase in the growth inhibition rate in experimental groups(P<0.05). The mean fluorescence intensity of RECK of control group and experimental groups(50,100,200μmol/L) were 0.15±0.01,0.21±0.02,0.25±0.02,0.29±0.01. And the IOD of RECK of these groups were 0.55±0.06,0.76±0.08,1.17±0.07,1.35±0.07. As compared with control group, NS398 caused a dose-dependent increase in the concentration of RECK in experimental groups (P<0.05). The mean fluorescence intensity of MT1-MMP of control group and experimental groups(50,100,200μmol/L) were 0.31±0.02,0.27±0.02,0.23±0.01,0.19±0.03.The IOD of MT1-MMP of these groups were 1.93±0.06,1.73±0.07,1.13±0.08,0.76±0.09. As compared with control group, NS398 caused a dose-dependent decrease in the concentration of MT1-MMP in experimental groups (P<0.05).Conclusion:The interactions between mononuclear cells and fibroblasts and H460 cells might increase the expression of MT1-MMP and MMP-2 activation in lung carcinoma cells, which can lead to the increase of extracellularmatrix followed by acceleration of tumor cellmigration and invasion. NS398 might increase the expression of RECK and decrease the expression of MT1-MMP in lung carcinoma cells, which can lead to the decrease of extracellularmatrix followed by acceleration of tumor cellmigration and invasion. It is probably an keymechanism of NS398 regulated lung carcinoma cellmigration.