TF/F Ⅶa complex induce the expression of MMP-7mRNA via P38 signal pathway in LOVO cells of colon cancer in vitro / 中国普通外科杂志

ABSTRACT

Objective To investigate the expression of matrix metalloproteinase 7(MMP-7) mRNA in LOVO cells of colon cancer induced by TF/F Ⅶ a and its signal pathway.Methods We transfected LOVO cells stably with RNAi plasmid targeting to tissue factor to get TFRNAi LOVO cells and detected efficiency of interference in TFRNAi LOVO cells based on Western blot analysis;Expression of MMP-7 was evaluated in LOVO cells treated with 100 nmol/L FⅦa in 0 h、4 h、8 h、12 h、24 h based on RT-PCR and Northern blot.Expression of MMP-7mRNA was determined in quiescent LOVO cells treated with different doses of FⅦa(0 nmol/L、10nmol/L、50 nmol/L、100 nmol/L、200 nmol/L)for 8 h based on Northern blot.Quiescent LOVO cells were treated for 0 h、4 h、8 h、12 h、16 h、24 h with 100 nmol/L FⅦa to evaluate the expression of p-P38;The expression level of MMP-7mRNA induced by 100 nmol/L FⅦa for 8 h in LOVO cells blocked by 10retool SB203580 0.5 h previously and in TFRNAi LOVO cells were measured by Northern blot.Results Northern blot analysis revealed that FⅦa markedly increased the expression of MMP-7mRNA in a time-and dose-dependent manner.Western blot analysis confirmed that FⅦa stimulates p-P38 in a time-dependent manner.SB203580 block 59.2% expression of MMP-7mRNA in LOVO cells induced by TF/FⅦa.In TFRNAi LOVO cells,the expression of MMP-7mRNA induced by TF/FⅦa was 48% less than that in normal LOVO cells.Conclusions TF/FⅦa Complex induces the expression of MMP-7mRNA in LOVO cells in vitro,possibly through P38 pathway.