Elevated expression of TGF-beta1 in head and neck cancer-associated fibroblasts.
Head and neck cancers are characterized by a vigorous desmoplastic response, but the contribution of stromal-derived growth factors to the tumor microenvironment is poorly understood. We evaluated the expression of stromal growth factor expression in head and neck squamous cell carcinoma (HNSCC) in normal and tumor-associated stromal cells. Stromal tissue was isolated from epithelial cells with laser capture microdissection (LCMD) and analyzed by cDNA array for the expression of TGFalpha, TGF-beta1, HGF, PDGF-alpha, IGFII, bFGF, aFGF, VEGFC, and VEGF. Primary fibroblasts were isolated in vitro from HNSCC tumors, adjacent histologically normal mucosa, and skin in vitro. Fibroblast populations were assessed for TGF-beta1 expression by ELISA and luciferase reporter assay to assess protein expression. We identified TGF-beta1 and IGFII overexpression in normal and tumor-associated stromal cells; however, only TGF-beta1 was significantly overexpressed (3.4-fold) in tumor-associated stroma. Assessment of carcinoma-associated fibroblasts (CAFs), normal dermal fibroblasts (NDFs), and normal mucosal fibroblasts (NMFs) in propagated fibroblasts demonstrated persistently elevated levels of TGF-beta1 in CAFs compared to NMF and NDF populations. Elevated levels of TGF-beta1 were identified in the stromal compartment of HNSCC tumors compared to normal mucosa by immunohistochemical analysis. These results suggest that TGF-beta1 mRNA and protein is specifically upregulated in CAFs in vitro and in vivo.
Rosenthal
Eben
E
Department of Surgery, Division of Otolaryngology-Head and Neck Surgery, University of Alabama at Birmingham, Birmingham, Alabama, USA.
McCrory
Allison
A
Talbert
Melissa
M
Young
Geoff
G
Murphy-Ullrich
Joanne
J
Gladson
Candece
C
D000818:Animals; D002294:Carcinoma, Squamous Cell / Q000235:genetics* / Q000473:pathology; D002478:Cells, Cultured; D016220:Fibroblast Growth Factor 1 / Q000235:genetics; D016222:Fibroblast Growth Factor 2 / Q000235:genetics; D005347:Fibroblasts / Q000502:physiology*; D005786:Gene Expression Regulation; D006258:Head and Neck Neoplasms / Q000235:genetics* / Q000473:pathology; D017228:Hepatocyte Growth Factor / Q000235:genetics; D006801:Humans; D007335:Insulin-Like Growth Factor II / Q000235:genetics; D009061:Mouth Mucosa / Q000166:cytology; D010982:Platelet-Derived Growth Factor / Q000235:genetics; D012016:Reference Values; D017154:Stromal Cells / Q000502:physiology; D016211:Transforming Growth Factor alpha / Q000235:genetics; D016212:Transforming Growth Factor beta / Q000235:genetics* / Q000378:metabolism; D053773:Transforming Growth Factor beta1; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics; D042582:Vascular Endothelial Growth Factor C / Q000235:genetics
D010982:Platelet-Derived Growth Factor; C508960:TGFB1 protein, human; D016211:Transforming Growth Factor alpha; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D042461:Vascular Endothelial Growth Factor A; D042582:Vascular Endothelial Growth Factor C; C075278:platelet-derived growth factor A; D016222:Fibroblast Growth Factor 2; D016220:Fibroblast Growth Factor 1; D017228:Hepatocyte Growth Factor; D007335:Insulin-Like Growth Factor II