The essential role of fibroblasts in esophageal squamous cell carcinoma-induced angiogenesis.
OBJECTIVE
Esophageal squamous cell carcinoma (ESCC) is known to be a highly angiogenic tumor. Here, we investigated the role of the stromal fibroblasts in the ESCC-induced angiogenic response using a novel 3-dimensional model.


METHODS
A novel assay was developed where cocultures of ESCC and esophageal fibroblasts induced human microvascular endothelial cell (HMVEC) vascular network formation in a 3-dimensional collagen gel. Biochemical studies showed that the ESCC-induced activation of the fibroblasts was required to induce vascular network formation via a transforming growth factor (TGF)-beta and vascular endothelial growth factor (VEGF)-dependent pathway.


RESULTS
Conditioned media from a panel of 4 ESCC lines transdifferentiated normal esophageal fibroblasts into myofibroblasts via TGF-beta signaling. The presence of fibroblasts was essential for efficient HMVEC network formation, and the addition of ESCC cells to these cultures greatly enhanced the angiogenic process. The role of TGF-beta in this process was shown by the complete inhibition of network formation following TGF-beta inhibitor treatment. Finally, we showed that ESCC-derived TGF-beta regulates angiogenesis through the release of VEGF from the fibroblasts and that the VEGF release was blocked following TGF-beta inhibition.


CONCLUSIONS
This study shows the essential role of fibroblasts in the ESCC angiogenic-induced response and suggests that the pharmacologic targeting of the TGF-beta signaling axis could be of therapeutic benefit in this deadly disease.
Noma
Kazuhiro
K
The Wistar Institute, Philadelphia, Pennsylvania 19104, USA.
Smalley
Keiran S M
KS
Lioni
Mercedes
M
Naomoto
Yoshio
Y
Tanaka
Noriaki
N
El-Deiry
Wafik
W
King
Alastair J
AJ
Nakagawa
Hiroshi
H
Herlyn
Meenhard
M
D001549:Benzamides / Q000494:pharmacology; D052117:Benzodioxoles / Q000494:pharmacology; D002294:Carcinoma, Squamous Cell / Q000098:blood supply / Q000378:metabolism*; D045744:Cell Line, Tumor; D054338:Cell Transdifferentiation; D018920:Coculture Techniques; D003094:Collagen / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D004938:Esophageal Neoplasms / Q000098:blood supply / Q000378:metabolism*; D004947:Esophagus / Q000196:embryology / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D005782:Gels; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D008833:Microcirculation / Q000378:metabolism; D009389:Neovascularization, Pathologic / Q000378:metabolism*; D019899:Paracrine Communication / Q000187:drug effects; D011720:Pyrazoles / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D014162:Transfection; D053773:Transforming Growth Factor beta1 / Q000037:antagonists & inhibitors / Q000378:metabolism; D053781:Transforming Growth Factor beta2 / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
C519132:2-(5-benzo(1,3)dioxol-5-yl-2-tert-butyl-3H-imidazol-4-yl)-6-methylpyridine hydrochloride; C509927:4-(4-(3-(pyridin-2-yl)-1H-pyrazol-4-yl)pyridin-2-yl)-N-(tetrahydro-2H-pyran-4-yl)benzamide; D001549:Benzamides; D052117:Benzodioxoles; D017077:Culture Media, Conditioned; D005782:Gels; D007093:Imidazoles; D011720:Pyrazoles; D011725:Pyridines; D053773:Transforming Growth Factor beta1; D053781:Transforming Growth Factor beta2; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D003094:Collagen