The expression of matrix metalloproteinase in pancreatic cancer cell line treated by antisense oligonucleotides of heparitinase in vitro / 中华内分泌外科杂志

ABSTRACT

Objective To investigate the expression of matrix metalloproteinase-2 (MMP-2)and matrix metalloproteinase-9(MMP-9) in MIA PaCa-2 cells blocked by AS-ODN cultured in hypoxia.Methods Heparanase(Hpa) expression of MIA PaCa-2 cells was blocked by AS-ODN and cultured in hypoxia.The expression of MMP-2 and MMP-9 mRNA and proteins in cell lysate was evaluated by RT-PCR and Western blot respectively,and the enzymatic activities of MMP-2 and MMP-9 in supernatants were detected by gelatinase activity assay.Results Hypoxia stimulated mRNA and protein expression of MMP-9 in cultured MIA PaCa-2 cells and elevated at 6h,12 h(P ＜0.05)and 24 h(P ＜ 0.01).When Hpa expression was inhibited by AS-ODN,the expression of MMP-9 mRNA and protein as well as the gelatinase activity in supernatant decreased dramatically at 12 h and 24 h,especially at 24h(P ＜0.01),however,no significant difference of MMP-2 expression and gelatinase activity was observed after AS-ODN transfection.(P ＞ 0.05).Conclusion In hypoxia,MMP-9 expression,either mRNA or protein in cultured MIA PaCa-2 cells,increased gradually accompanied with elevated gelatinase activities.When the heparanase expression was inhibited,the MMP-9 mRNA and protein,as well as the gelatinase B activity in supernatant,were decreased dramatically at 12h and 24h,however,no significantly differences of MMP-2 expression and gelatinase A activity were observed after the AS-ODN transfection.