1. Blocking mouse MMP-9 production in tumor cells and mouse cornea by short hairpin (sh) RNA encoding plasmids
- Author
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Barry T. Rouse, Susmit Suvas, Bumseok Kim, Yunsang Lee, Ahmet Kursat Azkur, and Udayasankar Kumaraguru
- Subjects
Biology ,Matrix Metalloproteinase Inhibitors ,medicine.disease_cause ,Small hairpin RNA ,Cornea ,Mice ,In vivo ,Cell Line, Tumor ,Chlorocebus aethiops ,Genetics ,medicine ,Animals ,Molecular Biology ,Gene knockdown ,Mice, Inbred BALB C ,Transfection ,Molecular biology ,In vitro ,Herpes simplex virus ,medicine.anatomical_structure ,Matrix Metalloproteinase 9 ,Cell culture ,COS Cells ,Colonic Neoplasms ,Keratitis, Herpetic ,Molecular Medicine ,RNA ,Female ,Plasmids - Abstract
In this study, we assessed the efficacy of the specific knockdown of matrix metalloprotein-9 (MMP-9) in vitro and in vivo using short hairpin RNA (shRNA) against MMP-9. Two plasmids were generated encoding shRNA (pshRNA) targeted against two distinct MMP-9 gene sequences. Transfection of these pshMMP-9s could be shown to specifically inhibit MMP-9 expression both in vivo and in vitro. The effect occurred in vitro both in cells that endogenously produce MMP-9 and in cells exogenously transfected with an MMP-9-encoding plasmid. Using an in vivo transfection approach, the pshMMP- 9 was also effective at inhibiting MMP-9 protein expression in the mouse cornea. pshMMP-9s were also tested against herpes simplex virus (HSV) in the cornea. Delivery of the pshMMP-9 stopped angiogenesis and decreased the severity of herpetic keratitis. However, interferon-alpha (IFN-alpha) and IFN- beta induced by pshRNA might also contribute to inhibition of herpetic simplex keratitis (HSK) in the cornea.
- Published
- 2005