The transforming growth factor-beta family members bone morphogenetic protein-2 and macrophage inhibitory cytokine-1 as mediators of the antiangiogenic activity of N-(4-hydroxyphenyl)retinamide.
OBJECTIVE
Tumor growth appears to be an angiogenesis-dependent process. N-(4-hydroxyphenyl)retinamide (fenretinide; 4HPR) has been found to inhibit and/or prevent tumor growth under diverse conditions. Although 4HPR is antiangiogenic, the molecular mechanisms of this effect remain largely unknown.


METHODS
Endothelial cells were treated with 4HPR in vitro to study the effects on migration, invasion, and organization, as well as gene expression by microarray and quantitative PCR studies. In vivo angiogenesis was evaluated in the Matrigel model.


RESULTS
4HPR treatment substantially modified the biological activities of endothelial cells, repressing their capacity to migrate, invade, and organize into capillary-like structures. The inhibition of invasion induced by 4HPR was also associated with decreased activities of the metalloproteases matrix metalloproteinase-2 and CD13/APN. Using oligonucleotide microarrays, we observed that bone morphogenetic protein-2 and macrophage inhibitory cytokine-1, two multifunctional cytokines of the transforming growth factor-beta family that regulate the growth, differentiation, apoptosis, and matrix accumulation of a variety of cells, are up-regulated in vitro by 4HPR. Both these molecules specifically inhibited endothelial cell growth, migration, and invasion in vitro and suppressed angiogenesis in the Matrigel plug assay in vivo. Blocking antibodies to bone morphogenetic protein-2 were able to reverse the suppressive effects of 4HPR in vitro and in vivo.


CONCLUSIONS
These data support the conclusion that 4HPR inhibits tumor growth by repression of new vessel growth and identify novel points of regulation of angiogenesis in transforming growth factor-beta family proteins.
Ferrari
Nicoletta
N
Molecular Oncology Laboratory, National Institute for Cancer Research, Genova, Italy.
Pfeffer
Ulrich
U
Dell'Eva
Raffaella
R
Ambrosini
Claudia
C
Noonan
Douglas M
DM
Albini
Adriana
A
D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003094:Collagen; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D004730:Endothelium, Vascular / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D017313:Fenretinide / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D055436:Growth Differentiation Factor 15; D006801:Humans; D007797:Laminin; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D045726:Metalloproteases / Q000378:metabolism; D051379:Mice; D009024:Morphogenesis / Q000187:drug effects; D041681:NIH 3T3 Cells; D020411:Oligonucleotide Array Sequence Analysis; D011509:Proteoglycans; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000235:genetics* / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology
D020533:Angiogenesis Inhibitors; C525718:BMP2 protein, human; C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D017077:Culture Media, Conditioned; D004338:Drug Combinations; C526140:GDF15 protein, human; C526141:Gdf15 protein, mouse; D055436:Growth Differentiation Factor 15; D007797:Laminin; D008565:Membrane Proteins; D011509:Proteoglycans; D016212:Transforming Growth Factor beta; C059485:matrigel; D017313:Fenretinide; D003094:Collagen; D045726:Metalloproteases