Extracellular
proteases have been shown to cooperatively influence matrix degradation and
tumor cell invasion through proteolytic cascades, with individual
proteases having distinct roles in
tumor growth, invasion, migration and angiogenesis. Matrix
metalloproteases (MMP)-9 and
cathepsin B have been shown to participate in the processes of
tumor growth, vascularization and invasion of
gliomas. In the present study, we used a cytomegalovirus promoter-driven
DNA template approach to induce hairpin
RNA (hpRNA)-triggered RNA interference (RNAi) to block MMP-9 and
cathepsin B gene expression with a single construct. Transfection of a plasmid vector-expressing
double-stranded RNA (dsRNA) for MMP-9 and
cathepsin B significantly inhibited MMP-9 and
cathepsin B expression and reduced the invasive behavior of SNB19,
glioblastoma cell line in
Matrigel and spheroid invasion models. Downregulation of MMP-9 and
cathepsin B using RNAi in SNB19 cells reduced cell-cell interaction of human microvascular endothelial cells, resulting in the disruption of capillary network formation in both in vitro and in vivo models. Direct intratumoral
injections of plasmid
DNA expressing hpRNA for MMP-9 and
cathepsin B significantly inhibited established
glioma tumor growth and invasion in intracranial
tumors in vivo. Further intraperitoneal (i.p.)
injections of plasmid
DNA expressing hpRNA for MMP-9 and
cathepsin B completely regressed pre-established
tumors for a long time (4 months) without any indication of these
tumor cells. For the first time, these observations demonstrate that the simultaneous RNAi-mediated targeting of MMP-9 and
cathepsin B has potential application for the treatment of human
gliomas.