title: Endothelial-mesenchymal transition in normal human esophageal endothelial cells cocultured with esophageal adenocarcinoma cells: role of IL-1β and TGF-β2.
abstract: Endothelial-mesenchymal transition (EndoMT) has been recognized as a key determinant of tumor microenvironment in cancer progression and metastasis. Endothelial cells undergoing EndoMT lose their endothelial markers, acquire the mesenchymal phenotype, and become more invasive with increased migratory abilities. Early stages of esophageal adenocarcinoma (EAC) are characterized by strong microvasculature whose impact in tumor progression remains undefined. Our aim was to determine the role of EndoMT in EAC by investigating the impact of tumor cells on normal primary human esophageal microvascular endothelial cells (HEMEC). HEMEC were either cocultured with OE33 adenocarcinoma cells or treated with IL-1β and transforming growth factor-β2 (TGF-β2) for indicated periods and analyzed for EndoMT-associated changes by real-time PCR, Western blotting, immunofluorescence staining, and functional assays. Additionally, human EAC tissues were investigated for detection of EndoMT-like cells. Our results demonstrate an increased expression of mesenchymal markers [fibroblast-specific protein 1 (FSP1), collagen1α2, vimentin, α-smooth muscle actin (α-SMA), and Snail], decreased expression of endothelial markers [CD31, von Willebrand factor VIII (vWF), and VE-cadherin], and elevated migration ability in HEMEC following coculture with OE33 cells. The EndoMT-related changes were inhibited by IL-1β and TGF-β2 gene silencing in OE33 cells. Recombinant IL-1β and TGF-β2 induced EndoMT in HEMEC. Although the level of VEGF expression was elevated in EndoMT cells, the angiogenic property of these cells was diminished. In vivo, by immunostaining EndoMT-like cells were detected at the invasive front of EAC. Our findings underscore a significant role for EndoMT in EAC and provide new insights into the mechanisms and significance of EndoMT in the context of tumor progression.
authors.affiliation: Department of Surgery, Medical College of Wisconsin, Milwaukee, Wisconsin;
authors.forename: Linghui
authors.initials: L
authors.lastname: Nie
authors.affiliation: Division of Gastroenterology/Hepathology, Medical College of Wisconsin, Milwaukee, Wisconsin; and.
authors.forename: Orestis
authors.initials: O
authors.lastname: Lyros
authors.affiliation: Department of Surgery, Medical College of Wisconsin, Milwaukee, Wisconsin;
authors.forename: Rituparna
authors.initials: R
authors.lastname: Medda
authors.affiliation: Division of Gastroenterology/Hepathology, Medical College of Wisconsin, Milwaukee, Wisconsin; and.
authors.forename: Nebojsa
authors.initials: N
authors.lastname: Jovanovic
authors.affiliation: Division of Gastroenterology/Hepathology, Medical College of Wisconsin, Milwaukee, Wisconsin; and.
authors.forename: Jamie L
authors.initials: JL
authors.lastname: Schmidt
authors.affiliation: Department of Surgery, Medical College of Wisconsin, Milwaukee, Wisconsin;
authors.forename: Mary F
authors.initials: MF
authors.lastname: Otterson
authors.affiliation: Department of Surgery, Medical College of Wisconsin, Milwaukee, Wisconsin;
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Johnson
authors.affiliation: Department of Pathology, Medical College of Wisconsin, Milwaukee, Wisconsin.
authors.forename: Behnaz
authors.initials: B
authors.lastname: Behmaram
authors.affiliation: Division of Gastroenterology/Hepathology, Medical College of Wisconsin, Milwaukee, Wisconsin; and.
authors.forename: Reza
authors.initials: R
authors.lastname: Shaker
authors.affiliation: Department of Surgery, Medical College of Wisconsin, Milwaukee, Wisconsin; prafiee@mcw.edu.
authors.forename: Parvaneh
authors.initials: P
authors.lastname: Rafiee
mesh_terms: D000230:Adenocarcinoma / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D004938:Esophageal Neoplasms / Q000378:metabolism / Q000473:pathology*; D004947:Esophagus / Q000166:cytology*; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism; D053583:Interleukin-1beta / Q000037:antagonists & inhibitors / Q000502:physiology*; D008648:Mesoderm / Q000166:cytology*; D053781:Transforming Growth Factor beta2 / Q000037:antagonists & inhibitors / Q000502:physiology*; D059016:Tumor Microenvironment; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D018836:Inflammation Mediators; D053583:Interleukin-1beta; D053781:Transforming Growth Factor beta2; D042461:Vascular Endothelial Growth Factor A
keywords: EAC; EndoMT; HEMEC; IL-1; TGF-β