Downregulation of matrix metalloproteinase-2 (MMP-2) utilizing adenovirus-mediated transfer of small interfering RNA (siRNA) in a novel spinal metastatic melanoma model

  • Authors: Andrew J. Tsung, Odysseas Kargiotis, Chandramu Chetty, Sajani S. Lakka, Meena Gujrati, Daniel G. Spomar, Dzung H. Dinh, Jasti S. Rao
  • View Affiliations

  • Published online on: Saturday, March 1, 2008
  • Pages: 557-564
  • DOI: 10.3892/ijo.32.3.557

Abstract

Matrix metalloproteinases (MMPs) comprise a class of secreted zinc-dependent endopeptidases implicated in the metastatic potential of tumor cells due to their ability to degrade the extracellular matrix (ECM) and basement membrane. Matrix metalloproteinase-2 (MMP-2) has been detected in high levels and correlates with invasiveness in human melanoma. We have studied the effect of adenovirus-mediated transfer of small interfering RNA (siRNA) against MMP-2 in the human melanoma cell line A2058. The delivery of these double-stranded RNA molecules represents an efficient technology in silencing disease-causing genes with known sequences at the post-transcriptional level. siRNA against MMP-2 mRNA (Ad-MMP-2) was found to decrease MMP-2 protein expression and activity in melanoma cells as demonstrated by western blotting and gelatin zymography. Furthermore, infection of cells with Ad-MMP-2 inhibited cellular migration and invasion as indicated by spheroid and matrigel assays. We also observed dose-dependent suppression of vascular network formation in an angiogenesis assay. Finally, we developed a nude mouse spinal metastatic model to investigate the local effects of tumor metastasis. Intravenous tail vein injection with Ad-MMP-2 on days 5, 9 and 11 after tumor implantation resulted in complete retention of neurological function as compared to control and scrambled vector (Ad-SV)-treated groups that showed complete paraplegia by day 14±2 days. Hematoxylin and eosin staining revealed decreased tumor size in the Ad-MMP-2-treated animals. This novel experimental model revealed that adenoviral-mediated transfer of RNA interference against MMP-2 results in the retention of neurological function and significantly inhibited tumor growth.
Journal Cover

March 2008
Volume 32 Issue 3

Print ISSN: 1019-6439
Online ISSN:1791-2423

2012 Impact Factor: 2.657
Ranked #31/196 Oncology
(total number of cites)

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
APA
Tsung, A., Kargiotis, O., Chetty, C., Lakka, S., Gujrati, M., Spomar, D., Dinh, D., & Rao, J. (2008). Downregulation of matrix metalloproteinase-2 (MMP-2) utilizing adenovirus-mediated transfer of small interfering RNA (siRNA) in a novel spinal metastatic melanoma model. International Journal of Oncology, 32(3), 557-564.
MLA
Tsung, Kargiotis, Chetty, Lakka, Gujrati, Spomar, Dinh, and Jasti Rao. "Downregulation of matrix metalloproteinase-2 (MMP-2) utilizing adenovirus-mediated transfer of small interfering RNA (siRNA) in a novel spinal metastatic melanoma model." International Journal of Oncology International Journal of Oncology 32.3 (2008): 557-564.
Chicago
Tsung, Kargiotis, Chetty, Lakka, Gujrati, Spomar, Dinh, and Jasti Rao. "Downregulation of matrix metalloproteinase-2 (MMP-2) utilizing adenovirus-mediated transfer of small interfering RNA (siRNA) in a novel spinal metastatic melanoma model." International Journal of Oncology International Journal of Oncology 32 no. 3 (2008): 557-564.