Blocking mouse MMP-9 production in tumor cells and mouse cornea by short hairpin (sh) RNA encoding plasmids.
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.
Department of Microbiology, University of Tennessee, Knoxville, 37996, USA.
Ahmet Kursat
AK
Azkur
Bumseok
B
Kim
Susmit
S
Suvas
Yunsang
Y
Lee
Udayasankar
U
Kumaraguru
Barry T
BT
Rouse
D000818:Animals; D019556:COS Cells; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D003110:Colonic Neoplasms / Q000201:enzymology* / Q000473:pathology; D003315:Cornea / Q000201:enzymology* / Q000821:virology; D005260:Female; D016849:Keratitis, Herpetic / Q000201:enzymology*; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis* / Q000235:genetics; D061965:Matrix Metalloproteinase Inhibitors; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids / Q000138:chemical synthesis / Q000502:physiology* / Q000627:therapeutic use; D012313:RNA / Q000138:chemical synthesis / Q000502:physiology* / Q000627:therapeutic use
D061965:Matrix Metalloproteinase Inhibitors; D012313:RNA; D020780:Matrix Metalloproteinase 9; C501830:Mmp9 protein, mouse