Effect of deoxyribozymes targeting c-Jun on solid tumor growth and angiogenesis in rodents.
BACKGROUND
The basic region-leucine zipper protein c-Jun has been linked to cell proliferation, transformation, and apoptosis. However, a direct role for c-Jun in angiogenesis has not been shown.


METHODS
We used human microvascular endothelial cells (HMEC-1) transfected with a DNAzyme targeting the c-Jun mRNA (Dz13), related oligonucleotides, or vehicle in in vitro models of microvascular endothelial cell proliferation, migration, chemoinvasion, and tubule formation, a rat model of corneal neovascularization, and a mouse model of solid tumor growth and vascular endothelial growth factor (VEGF)-induced angiogenesis. All statistical tests were two-sided.


RESULTS
Compared with mock-transfected cells, HMEC-1 cells transfected with Dz13 expressed less c-Jun protein and possessed lower DNA-binding activity. Dz13 blocked endothelial cell proliferation, migration, chemoinvasion, and tubule formation. Dz13 inhibited the endothelial cell expression and proteolytic activity of MMP-2, a c-Jun-dependent gene. Dz13 inhibited VEGF-induced neovascularization in the rat cornea compared with vehicle control (Dz13 versus vehicle: 4.0 neovessels versus 30.7 neovessels, difference = 26.7 neovessels; P =.004; area occupied by new blood vessels for Dz13 versus vehicle: 0.35 mm2 versus 1.52 mm2, difference = 1.17 mm2; P =.005) as well as solid melanoma growth in mice (Dz13 versus vehicle at 14 days: 108 mm3 versus 283 mm3, difference = 175 mm3; P =.006) with greatly reduced vascular density (Dz13 versus vehicle: 30% versus 100%, difference = 70%; P<.001).


CONCLUSIONS
DNAzymes targeting c-Jun may have therapeutic potential as inhibitors of tumor angiogenesis and growth.
Zhang
Guishui
G
Centre for Vascular Research, The University of New South Wales and Department of Haematology, The Prince of Wales Hospital, Sydney, Australia.
Dass
Crispin R
CR
Sumithran
Eric
E
Di Girolamo
Nick
N
Sun
Lun-Quan
LQ
Khachigian
Levon M
LM
D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D015153:Blotting, Western; D002455:Cell Division; D002460:Cell Line; D002465:Cell Movement; D003315:Cornea / Q000098:blood supply; D021881:DNA, Catalytic / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D042783:Endothelial Cells; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D016758:Genes, jun / Q000187:drug effects; D006801:Humans; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D061965:Matrix Metalloproteinase Inhibitors; D008546:Melanoma, Experimental / Q000188:drug therapy* / Q000201:enzymology; D051379:Mice; D008810:Mice, Inbred C57BL; D009389:Neovascularization, Pathologic / Q000188:drug therapy* / Q000201:enzymology; D016755:Proto-Oncogene Proteins c-jun / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A
D020533:Angiogenesis Inhibitors; D000970:Antineoplastic Agents; D021881:DNA, Catalytic; C485475:Dz13 DNAzyme; D061965:Matrix Metalloproteinase Inhibitors; D016755:Proto-Oncogene Proteins c-jun; D042461:Vascular Endothelial Growth Factor A; D020778:Matrix Metalloproteinase 2