Inhibition of invasion, angiogenesis, tumor growth, and metastasis by adenovirus-mediated transfer of antisense uPAR and MMP-9 in non–small cell lung cancer cells

Abstract

Lung cancer is currently the leading cause of cancer deaths in the United States. Conventional therapeutic treatments, including surgery, chemotherapy, and radiation therapy, have achieved only limited success. The overexpression of proteases, such as urokinase-type plasminogen activator (uPA), its receptor (uPAR), and matrix metalloproteinases (MMP), is correlated with the progression of lung cancer. In the present study, we used a replication-deficient adenovirus capable of expressing antisense uPAR and antisense MMP-9 transcripts to simultaneously down-regulate uPAR and MMP-9 in H1299 cells. Ad-uPAR-MMP-9 infection of H1299 cells resulted in a dose-and time-dependent decrease of uPAR protein levels and MMP-9 activity as determined by Western blotting and gelatin zymography, respectively. Corresponding immunohistochemical analysis also showed that Ad-uPAR-MMP-9 infection inhibited uPAR and MMP-9 expression. As shown by Boyden chamber assay, Ad-uPAR-MMP-9 infection significantly decreased the invasive capacity of H1299 cells compared with mock and Ad-CMV (empty vector) -infected cells in vitro . Furthermore, Ad-uPAR-MMP-9 infection inhibited capillary-like structure formation in H1299 cells cocultured with endothelial cells in a dose-dependent manner compared with mock-and Ad-CMV-infected cells. Ad-uPAR-MMP-9 injection caused the regression of s.c. induced tumors after s.c. injection with H1299 lung cancer cells and inhibited lung metastasis in the metastatic model with A549 cells. These data suggest that Ad-uPAR-MMP-9 shows its antitumor activity against both established and early phases of lung cancer metastases by causing the destruction of the tumor vasculature. In summary, adenovirus-mediated inhibition of uPA-uPAR interaction and MMP-9 on the cell surface may be a promising anti-invasion and antimetastatic strategy for cancer gene therapy. [Mol Cancer Ther 2005;4(9):1399 -408]

Fibronectin Increases Matrix Metalloproteinase 9 Expression through Activation of c-Fos via Extracellular-regulated Kinase and Phosphatidylinositol 3-Kinase Pathways in Human Lung Carcinoma Cells

“…Homogeneous MMP-9 expression in cancer cells is an independent prognostic factor in operable NSCLC, so MMP-9 may be considered as target for adjuvant anticancer therapy in operable NSCLC using selective MMP inhibitors with high specificity for MMP-9. MMP-9 inhibitors and an adenovirus-mediated transfer of antisense MMP-9 have been shown to inhibit invasion, angiogenesis, growth, and metastasis in NSCLC cells (29,30). In human lung cancer cells, high expression of MMP-9 is associated with growth, metastasis, and progression, and blockade of MMP-9 has been shown to control lymph node metastasis and prolong the life span of lung cancer patients (29 -32).…”

Urokinase-type Plasminogen Activator Receptor (uPAR)-mediated Regulation of WNT/β-Catenin Signaling Is Enhanced in Irradiated Medulloblastoma Cells

“…Furthermore, the role of uPAR in angiogenesis, inflammation, wound repair, and tumor progression/metastasis (42)(43)(44)(45) is due to its ability to engage in multiple protein/protein interactions (46 -50). Because uPAR is devoid of a transmembrane and cytoplasmic domain, we anticipated that association of uPAR with ␤-catenin would involve other interlinking molecules.…”

Urokinase and its receptors in chronic kidney disease

“…Pharmaceutical strategies of uPAR inactivation or blockade for in vivo evaluation have included RNA interference (85), adenovirus-mediated transfer of anti-sense uPAR (138), uPAR signaling blocked with anti-sense oligonucleotides or other small molecule inhibitors (139), adenovirus-mediated delivery of an uPA/uPAR antagonist (107), inhibitory monoclonal antibodies (140), and thalidomide (141). Soluble uPAR has also been investigated as a potential inhibitor of tumor growth (142).…”

“…Soluble uPAR has also been investigated as a potential inhibitor of tumor growth (142). These intervention strategies have been shown to decrease tumor invasion/metastasis, angiogenesis and growth in breast cancer, ovarian cancer, brain tumor, gliomas, glioblastoma multiform, osteosarcoma, nonsmall cell lung cancer, prostate cancer, and melanoma (85,138,139,141,142); to reduce retinal neovascularization in retinopathy (107,143); and to suppress macrophage infiltration into the vascular wall of ApoE-deficient mice (144).…”