title: TGFβ loss activates ADAMTS-1-mediated EGF-dependent invasion in a model of esophageal cell invasion.
abstract: The TGFβ signaling pathway is essential to epithelial homeostasis and is often inhibited during progression of esophageal squamous cell carcinoma. Recently, an important role for TGFβ signaling has been described in the crosstalk between epithelial and stromal cells regulating squamous tumor cell invasion in mouse models of head-and-neck squamous cell carcinoma (HNSCC). Loss of TGFβ signaling, in either compartment, leads to HNSCC however, the mechanisms involved are not well understood. Using organotypic reconstruct cultures (OTC) to model the interaction between epithelial and stromal cells that occur in dysplastic lesions, we show that loss of TGFβ signaling promotes an invasive phenotype in both fibroblast and epithelial compartments. Employing immortalized esophageal keratinocytes established to reproduce common mutations of esophageal squamous cell carcinoma, we show that treatment of OTC with inhibitors of TGFβ signaling (A83-01 or SB431542) enhances invasion of epithelial cells into a fibroblast-embedded Matrigel/collagen I matrix. Invasion induced by A83-01 is independent of proliferation but relies on protease activity and expression of ADAMTS-1 and can be altered by matrix density. This invasion was associated with increased expression of pro-inflammatory cytokines, IL1 and EGFR ligands HB-EGF and TGFα. Altering EGF signaling prevented or induced epithelial cell invasion in this model. Loss of expression of the TGFβ target gene ROBO1 suggested that chemorepulsion may regulate keratinocyte invasion. Taken together, our data show increased invasion through inhibition of TGFβ signaling altered epithelial-fibroblasts interactions, repressing markers of activated fibroblasts, and altering integrin-fibronectin interactions. These results suggest that inhibition of TGFβ signaling modulates an array of pathways that combined promote multiple aspects of tumor invasion.
authors: Department of Surgery, Vanderbilt University, Nashville, TN, USA.
authors: Grégoire F
authors: GF
authors: Le Bras
authors: Department of Surgery, Vanderbilt University, Nashville, TN, USA.
authors: Chase
authors: C
authors: Taylor
authors: Department of Surgery, Vanderbilt University, Nashville, TN, USA.
authors: Rainelli B
authors: RB
authors: Koumangoye
authors: Department of Pathology, Vanderbilt University, Nashville, TN, USA.
authors: Frank
authors: F
authors: Revetta
authors: Department of Cancer Biology, Vanderbilt University, Nashville, TN, USA.
authors: Holli A
authors: HA
authors: Loomans
authors: Department of Surgery, Vanderbilt University, Nashville, TN, USA; Department of Cancer Biology, Vanderbilt University, Nashville, TN, USA; Department of Vanderbilt Ingram Cancer Center, Vanderbilt University, Nashville, TN, USA. Electronic address: claudia.andl@vanderbilt.edu.
authors: Claudia D
authors: CD
authors: Andl
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism*; D000071097:ADAMTS1 Protein; D002294:Carcinoma, Squamous Cell / Q000378:metabolism*; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D004938:Esophageal Neoplasms / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism / Q000502:physiology; D066257:Heparin-binding EGF-like Growth Factor / Q000378:metabolism*; D006801:Humans; D007375:Interleukin-1 / Q000378:metabolism; D015603:Keratinocytes / Q000378:metabolism* / Q000502:physiology; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D011971:Receptors, Immunologic / Q000235:genetics / Q000378:metabolism; D018125:Receptors, Transforming Growth Factor beta / Q000037:antagonists & inhibitors; D016211:Transforming Growth Factor alpha / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D000097905:Roundabout Proteins
chemical_list: D051722:ADAM Proteins; D000071097:ADAMTS1 Protein; D066257:Heparin-binding EGF-like Growth Factor; D007375:Interleukin-1; D009419:Nerve Tissue Proteins; D011971:Receptors, Immunologic; D018125:Receptors, Transforming Growth Factor beta; D016211:Transforming Growth Factor alpha; D016212:Transforming Growth Factor beta; D000097905:Roundabout Proteins; C103859:ADAMTS1 protein, human
keywords: ADAMTS-1; Cell matrix interaction; ESCC; Invasion; MMP14