Abstract
Androgen deprivation therapy causes a paradoxical elevation of matrix metalloproteinases (MMPs) including MMP-9 resulting in aggressive tumor phenotype in many patients with prostate cancer. In this study, we have evaluated a novel antisense phosphorodiamidate Morpholino oligomer (PMO) targeted against MMP-9 in models of angiogenesis and in human prostate xenograft in athymic mice. The treatment of androgen-independent DU145 human prostate cells with a 21-mer MMP-9 antisense PMO caused a dose-dependent inhibition of cell proliferation compared to scrambled or MMP-2 antisense PMO at similar concentrations. This was associated with decreases in MMP-9 expression, gelatinolytic activity and increased stability of the insulin-like growth factor-binding protein (IGFBP-3), a proapoptotic factor and MMP-9 substrate. In vitro invasion assays revealed a 40–60% inhibition of DU145 cell invasion in the presence of 25 μM MMP-9 antisense PMO. A significant decrease in endothelial cell migration and vascularization was observed in the Matrigel plug assay in mice when treated intraperitoneally with 300 μg/day MMP-9 antisense for 21 days. In the highly vascular DU145 tumor xenografts, MMP-9 inhibition caused decreased tumor growth with regression in 50% of the animals. Histological analysis revealed increased apoptosis and fibrous tissue deposits in the MMP-9 antisense-treated tumors compared to the scrambled and saline controls. No apparent toxicity or mortality was associated with the MMP-9 PMO treatment. In summary, the MMP-9 antisense PMO inhibited in vitro prostate cancer cell proliferation, invasion and in vivo angiogenesis. These data establish the feasibility of developing a site-directed, nontoxic antisense therapeutic agent for inhibiting local invasion and metastasis.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Greenlee RT, Hill-Harmon MB, Murray T, Thun M . Cancer statistics (2001). CA: Cancer J Clin. 2001;51:15–36.
Richie JP . Anti-androgens and other hormonal therapies for prostate cancer. Urology. 1999;54:15–18.
Devi GR . Prostate cancer: status of current treatments and emerging antisense-based therapies. Curr Opin Mol Ther. 2002;4:138–148.
Sadar MD, Hussain M, Bruchovsky N . Prostate cancer: molecular biology of early progression to androgen in-dependence. Endocrin Rel Cancer. 1999;6:487–502.
Overall CM . Molecular determinants of metalloproteinase substrate specificity: matrix metalloproteinase substrate binding domains, modules, and exosites. Mol Biotechnol. 2002;22:51–86.
Creemers EE, Cleutjens JP, Smits JF, Daemen MJ . Matrix metalloproteinase inhibition after myocardial infarction: a new approach to prevent heart failure? Circ Res. 2001;89:201–210 (Review).
Porter KE, Turner NA . Statins for the prevention of vein graft stenosis: a role for inhibition of matrix metalloproteinase-9. Biochem Soc Trans. 2002;30:120–126.
Lijnen HR . Plasmin and matrix metalloproteinases in vascular remodeling. Thromb Haemost. 2001;86:324–333. (Review).
Harada H, Furuya M, Ishikura H, Shindo J, Koyanagi T, Yoshiki T . Expression of matrix metalloproteinase in the fluids of renal cystic lesions. J Urol. 2002;168:19–22.
Faber-Elmann A, Sthoeger Z, Tcherniack A, Dayan M, Mozes E . Activity of matrix metalloproteinase-9 is elevated in sera of patients with systemic lupus erythematosus. Clin Exp Immunol. 2002;127:393–398.
Giannelli G, Antonaci S . Gelatinases and their inhibitors in tumor metastasis: from biological research to medical applications. Histol Histopathol. 2002;17:339–345.
Egeblad M, Werb Z . New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer. 2002;2:161–174.
Pajouh MS, Nagle RB, Breathnach R, Finch JS, Brawer MK, Bowden GT . Expression of metalloproteinase genes in human prostate cancer. J Cancer Res Clin Oncol. 1991;117:144–150.
Lokeshwar BL, Selzer MG, Block NL, Gunja-Smith Z . Secretion of matrix metalloproteinases and their inhibitors (tissue inhibitor of metalloproteinases) by human prostate in explant cultures: reduced tissue inhibitor of metalloproteinase secretion by malignant tissues. Cancer Res. 1993;53:4493–4498.
Stearns ME, Wang M . Type IV collagenase (M(r) 72,000) expression in human prostate: benign and malignant tissue. Cancer Res. 1993;53:878–883.
Sternlicht MD, Werb Z . How matrix metalloproteinases regulate cell behavior. Annu Rev Cell Dev Biol. 2001;17:463–516.
Rajah R, Valentinis B, Cohen P . IGFBP-3 induces apoptosis and mediates the effects of transforming growth factor-b1 on programmed cell death through a p53- and IGF-independent mechanism. J Biol Chem. 1997;272:12181–12188.
Devi GR, Sprenger C, Graham D, Plymate S, Rosenfeld RG . Overexpression of IGFBP-3 induces cell growth inhibition and early apoptosis in malignant human prostate cancer cells. Prostate 2002;53:200–210.
Cohen P, Peehl D, Graves H, Rosenfeld R . Biological effects of prostate specific antigen (PSA) as an IGF binding protein-3 (IGFBP-3) protease. J Endocrinol. 1994;142:407–415.
Smith GL, Doherty AP, Mitchell H, Hanham IW, Christmas TJ, Epstein RJ . Inverse relation between prostate-specific antigen and insulin-like growth factor-binding protein 3 in bone metastases and serum of patients with prostate cancer. Lancet. 1999;354:2053–2054.
Shariat SF, Lamb DJ, Kattan MW et al. Association of preoperative plasma levels of insulin-like growth factor 1 and insulin-like growth factor binding proteins-2 and -3 with prostate cancer invasion, progression, and metastasis. J Clin Oncol. 2002;20:833–841.
Zucker S, Cao J, Chen WT . Critical appraisal of the use of matrix metalloproteinase inhibitors in cancer treatment. Oncogene. 2000;19:6642–6650.
Overall CM, Lopez-Otin C . Strategies for MMP inhibition in cancer: innovations for the post-trial era. Nat Rev Cancer. 2002;2:657–672.
Summerton J, Weller D . Morpholino antisense oligomers: design, preparation, and properties. Antisense Nucleic Acids Drug Dev. 1997;7:187–195.
Hudziak RM, Summerton J, Weller DD, Iversen PL . Antiproliferative effects of steric blocking phosphorodiamidate morpholino antisense agents directed against c-myc. Antisense Nucleic Acids Drug Dev. 2000;10:163–176.
Devi GR, Oldenkamp JR, London CA, Iversen PL . Inhibition of human chorionic gonadotropin beta subunit modulates the mitogenic effect of c-myc in human prostate cancer cells. Prostate 2002;53:200–210.
Attiga FA, Fernandez PM, Weeraratna AT, Manyak MJ, Patierno SR . Inhibitors of prostaglandin synthesis inhibit human prostate tumor cell invasiveness and reduce the release of matrix metalloproteinases. Cancer Res. 2000;60:4629–4637.
Smith JP, Verderame MF, Zagon IS . Antisense oligonucleotides to gastrin inhibit growth of human pancreatic cancer. Cancer Lett. 1999;135:107–112.
Sauter ER, Takemoto R, Litwin S, Herlyn M . p53 alone or in combination with antisense cyclin D1 induces apoptosis and reduces tumor size in human melanoma. Cancer Gene Ther. 2002;9:807–812.
Iversen PL, Arora V, Acker AJ, Mason D, Devi GR . Efficacy of antisense Morpholino oligomer targeted to c-myc in prostate cancer xenograft murine model and a phase I safety study in humans. Clin Cancer Res. 2003;9:2510–2519.
Desai SB, Libutti SK . Tumor angiogenesis and endothelial cell modulatory factors. J Immunother. 1999;22:186–211.
Folkman J . Angiogenesis-dependent diseases. Semin Oncol. 2001;28:536–542.
Lokeshwar BL . MMP inhibition in prostate cancer. Ann NY Acad Sci. 1999;878:271–289 (Review).
Manes S, Llorente M, Lacalle RA, Gomez-Mouton C, Kremer L, Mira E, Martinez- AC . The matrix metalloproteinase-9 regulates the insulin-like growth factor-triggered autocrine response in DU-145 carcinoma cells. J Biol Chem. 1999;274:6935–6945.
Angelloz-Nicoud P, Binoux M . Autocrine regulation of cell proliferation by the insulin-like growth factor (IGF) and IGF binding protein-3 protease system in a human prostate carcinoma cell line (PC-3). Endocrinology. 1995;136:5485–5492.
Devi GR, Yang DY, Rosenfeld RG, Oh Y . Differential effects of IGFBP-3 and its proteolytic fragments on ligand binding, IGFIR signaling and cell surface association. Endocrinology. 2000;141:4171–4179.
Devi GR, Oh Y, Rosenfeld RG . Effect of IGFBP-3 on IGF- and IGF-analog-induced IGFIR signaling. Growth Horm. IGF Res. 2001;11:231–239.
Summerton J, Weller D . Antisense properties of morpholino oligomers. Nucleosides Nucleotides. 1997;16:889–898.
Summerton J . Morpholino antisense oligomers: the case for an RNase H-independent structural type. Biochim Biophys Acta. 1999;1489:141–158.
Iversen PL . PMO: favorable properties for sequence-specific inactivation. Curr Opin Mol Ther. 2001;3:235–238.
Giles RV, Spiller DG, Clark RE, Tidd DM . Antisense morpholino oligonucleotide analog induces missplicing of c-myc mRNA. Antisense Nucleic Acids Drug Dev. 1999;9:213–220.
Ghosh C, Stein D, Weller D, Iversen PL . Evaluation of antisense mechanisms of action. Methods Enzymol. 2000;313:135.
Knapp DC, Mata JE, Reddy MT, Devi GR, Iversen PL . Resistance to chemotherapeutic drugs overcome by c-Myc inhibition in Lewis lung carcinoma murine model. Anti-Cancer Drugs. 2003;14:39–47.
Arora V, Knapp DC, Reddy MT, Weller DD, Iversen PL . Bioavailability and efficacy of antisense morpholino oligomers targeted to c-Myc and cytochrome P-450 3A2 following oral administration in rats. J Pharm Sci. 2002;91:1009–1017.
Arora V, Hannah TL, Iversen PL, Brand R . Transdermal use of phosphorodiamidate Morpholino oligomer AVI-4472 inhibits cytochrome P450 3A2 activity in male rats. Pharm Res. 2002;19:1465–1470.
Henry SP, Novotny W, Leeds J, Auletta C, Kornbrust DJ . Inhibition of coagulation by a phosphorothioate oligonucleotide. Antisense Nucleic Acids Drug Dev. 1997;7:503–510.
Iversen PL, Cornish KG, Iversen LJ, Mata JE, Bylund DB . Bolus intravenous injection of phosphorothioate oligonucleotides causes hypotension by acting as alpha1-adrenergic receptor antagonists. Toxicol Appl Pharm. 1999;160:289–296.
Levin AA . A review of the issues in the pharmacokinetics and toxicology of phosphorothioate antisense oligonucleotides. Biochem Biophys Acta. 1999;1489:69–84.
Acknowledgements
This work was supported by a grant from Department of Defense-DAMD17-01-1-0017 (GRD) and funds from AVI BioPharma. We thank Melissa Cate and Andrew Kroecker for technical assistance; Dr Ojerio (Oregon State University) for expert advice on animal care issues; and Anita Sonn (Oregon State University) for assistance in preparation of slides. We also acknowledge the technical assistance provided by the synthesis and purification group at AVI BioPharma.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
London, C., Sekhon, H., Arora, V. et al. A novel antisense inhibitor of MMP-9 attenuates angiogenesis, human prostate cancer cell invasion and tumorigenicity. Cancer Gene Ther 10, 823–832 (2003). https://doi.org/10.1038/sj.cgt.7700642
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.cgt.7700642
Keywords
This article is cited by
-
The effects of resveratrol on the expression of VEGF, TGF-β, and MMP-9 in endometrial stromal cells of women with endometriosis
Scientific Reports (2021)
-
Prognostic significance of the tumour-adjacent tissue in head and neck cancers
Tumor Biology (2015)
-
Prostate cancer chemoprevention in men of African descent: current state of the art and opportunities for future research
Cancer Causes & Control (2013)
-
Oligonucleotide suppression of bcl-2 in LNCaP cells is compensated by increased androgen sensitivity, p53 and oncogene activity, and suppressed caspase-3
Medical Oncology (2013)
-
Bone morphogenetic protein 4 (BMP4) is required for migration and invasion of breast cancer
Molecular and Cellular Biochemistry (2012)