TGFbeta1 antagonistic peptides inhibit TGFbeta1-dependent angiogenesis.
The role of transforming growth factor beta (TGFbeta) in tumor promotion and in angiogenesis is context-dependent. While TGFbeta prevents tumor growth and angiogenesis in early phases of tumor development, evidence is accumulating about its pro-angiogenic and tumor promotion activities in late-stages of tumor progression. Here we have studied, in an experimental context previously reported to disclose the pro-angiogenic effects of TGFbeta, the blocking activity of TGFbeta antagonist peptides. In agreement with previous results, we have observed that TGFbeta exerts a powerful pro-angiogenic activity on human normal dermal microvascular endothelial cells (MVEC), by promoting invasion and capillary morphogenesis in Matrigel. No apoptotic activity of TGFbeta was observed. By RT-PCR we have shown that TGFbeta up-regulates expression not only of plasminogen activator inhibitor type-1 (PAI-1), but also of the urokinase-type plasminogen activator receptor (uPAR), whose inhibition by specific antibodies blunted the TGFbeta angiogenic response in vitro. The SMAD2/3 and FAK signaling pathways were activated by TGFbeta in MVEC, as an early and late response, respectively. The use of two different TGFbeta1 antagonist peptides, derived from TGFbeta type III receptor sequence and 15-mer phage display technology, inhibited the signaling and pro-angiogenic response in vitro, as well as uPAR and PAI-1 up-regulation of MVEC following TGFbeta challenge. The anti-angiogenic properties of both inhibitors were evident also in the in vivo TGFbeta Matrigel Sponge Assay. These results may be relevant to develop a potentially fruitful strategy for the therapy of late-stage-associated tumor angiogenesis.
Serratì
Simona
S
Department of Experimental Pathology and Oncology, Center for the Study at Molecular and Clinical Level of Chronic, Degenerative and Neoplastic Diseases to DEvelop NOvel THEerapies, University of Florence, Viale G.B. Morgagni, 50, 50134 Florence, Italy.
Margheri
Francesca
F
Pucci
Marco
M
Cantelmo
Anna Rita
AR
Cammarota
Rosaria
R
Dotor
Javier
J
Borràs-Cuesta
Francisco
F
Fibbi
Gabriella
G
Albini
Adriana
A
Del Rosso
Mario
M
D001483:Base Sequence; D015153:Blotting, Western; D002196:Capillaries / Q000254:growth & development; D049109:Cell Proliferation / Q000187:drug effects; D017931:DNA Primers; D006801:Humans; D007150:Immunohistochemistry; D009389:Neovascularization, Pathologic / Q000517:prevention & control*; D010455:Peptides / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053773:Transforming Growth Factor beta1 / Q000037:antagonists & inhibitors* / Q000502:physiology
D017931:DNA Primers; D010455:Peptides; C508960:TGFB1 protein, human; D053773:Transforming Growth Factor beta1