Inhibition of cathepsin B and MMP-9 gene expression in glioblastoma cell line via RNA interference reduces tumor cell invasion, tumor growth and angiogenesis

Abstract

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 vectorexpressing 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 (ip) 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.

RNA Interference-directed Knockdown of Urokinase Plasminogen Activator and Urokinase Plasminogen Activator Receptor Inhibits Prostate Cancer Cell Invasion, Survival, and Tumorigenicity in Vivo

“…Several laboratories are currently testing the feasibility of RNAi-mediated gene silencing as a novel tool for arresting tumor growth and killing cancer cells, and the initial results are promising (35,40,54,55). Our recent studies demonstrate the potential of simultaneous inhibition of two genes using a plasmid-based siRNA system (40,57). In this study, we have used plasmid-based RNAi to target uPA and uPAR genes, which are markedly overexpressed in a wide variety of human cancers, including prostate cancer, singly and simultaneously.…”

CXCL12/CXCR4 Transactivates HER2 in Lipid Rafts of Prostate Cancer Cells and Promotes Growth of Metastatic Deposits in Bone

“…Furthermore, these data suggest that CXCL12/CXCR4 transactivation of HER2 is an upstream event of Akt activation in prostate cancer cells. Several studies further suggest that MMP-9 plays a key role in tumor cell migration and invasion by remodeling the matrix; this fact is further supported by MMP-9 knockdown studies (41)(42)(43). In prostate cancer bone metastasis, MMP-9 plays a key role in the initial establishment of metastatic formation by fostering the tumor cell -induced pathophysiologic matrix remodeling and subsequent expansion of tumor deposits in bone tissue (42).…”

The mannose 6‐phosphate/insulin‐like growth factor II receptor restricts the tumourigenicity and invasiveness of squamous cell carcinoma cells

“…45,46 Hence, our results are in good agreement with recent in vivo studies, which provided strong evidence that cysteine cathepsins participate in tumour invasion. [47][48][49] Therefore, it is possible that M6P/IGF2R restricts tumour invasion not only by dampening the biological activity of IGF-II, but also by blocking the pericellular accumulation of matrix-degrading proteinases such as cysteine cathepsins.…”