Adenoviral mediated interferon-alpha 2b gene therapy suppresses the pro-angiogenic effect of vascular endothelial growth factor in superficial bladder cancer.
OBJECTIVE
Intravesical adenovirus mediated interferon-alpha gene transfer has a potent therapeutic effect against superficial human bladder carcinoma xenografts growing in the bladder of athymic nude mice. We determined whether the inhibition of angiogenesis might contribute to the antitumor effect.


METHODS
We treated several human urothelial carcinoma cells with adenovirus mediated interferon-alpha 2b and monitored its effects on the production of angiogenic factors using real-time reverse-transcription polymerase chain reaction, Western blotting, and immunohistochemical analysis and a gel shift based transcription factor array. To assess the role of adenovirus mediated interferon 2b in angiogenic activity we used in vitro invasion assays and evaluated the anti-angiogenic effects of adenovirus mediated interferon gene therapy in an orthotopic murine model of human superficial bladder cancer.


RESULTS
In adenovirus mediated interferon-alpha infected 253J B-V cells vascular endothelial growth factor was decreased and anti-angiogenic interferon-gamma inducible protein 10 was up-regulated. In contrast, the addition of as much as 100,000 IU recombinant interferon had no apparent effect on vascular endothelial growth factor production. Conditioned medium derived from adenovirus mediated interferon 2b infected 253J B-V cells greatly decreased the invasive potential of human endothelial cells and down-regulated their matrix metalloproteinase 2 expression compared to controls. Furthermore, adenovirus mediated interferon 2b blocked pro-angiogenic nuclear signals, such as the transcription factors activating protein-1 and 2, stimulating protein-1, nuclear factor kappaB and c-myb. In vivo experiments revealed significant vascular endothelial growth factor down-regulation and decreased tumor vessel density in the adenovirus mediated interferon 2b treated group compared to controls.


CONCLUSIONS
Treatment with adenovirus mediated interferon 2b increases the angiostatic activity of the bladder cancer microenvironment. This inhibition may prove beneficial for treating superficial bladder cancer with adenovirus mediated interferon-alpha and hopefully contribute to a decreased recurrence rate of this neoplasm.
Adam
Liana
L
Department of Urology, University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030, USA. ladam@mdanderson.org
Black
Peter C
PC
Kassouf
Wassim
W
Eve
Beryl
B
McConkey
David
D
Munsell
Mark F
MF
Benedict
William F
WF
Dinney
Colin P N
CP
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000970:Antineoplastic Agents / Q000627:therapeutic use*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D007150:Immunohistochemistry; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000627:therapeutic use*; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis / Q000235:genetics; D061965:Matrix Metalloproteinase Inhibitors; D051379:Mice; D008819:Mice, Nude; D018613:Microscopy, Confocal; D016328:NF-kappa B / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics; D009389:Neovascularization, Pathologic / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control; D020598:Proto-Oncogene Proteins c-myb / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D012334:RNA, Neoplasm / Q000187:drug effects / Q000235:genetics*; D011994:Recombinant Proteins; D018808:Transcription Factor AP-1 / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics; D050656:Transcription Factor AP-2 / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics; D001749:Urinary Bladder Neoplasms / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics*
D000970:Antineoplastic Agents; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D061965:Matrix Metalloproteinase Inhibitors; D016328:NF-kappa B; D020598:Proto-Oncogene Proteins c-myb; D012334:RNA, Neoplasm; D011994:Recombinant Proteins; D018808:Transcription Factor AP-1; D050656:Transcription Factor AP-2; D042461:Vascular Endothelial Growth Factor A; D020778:Matrix Metalloproteinase 2