Endothelium-dependent epithelial–mesenchymal transition of tumor cells: Exclusive roles of transforming growth factor β1 and β2

Abstract Background Induction of epithelial–mesenchymal transition (EMT) is essential for the metastasis of tumor cells and maintaining their stemness. This study aimed to examine whether endothelial cells, which are most closely located to tumor cells in vivo , play a role in inducing EMT in tumor cells or not. Methods Concentrated culture medium of bovine aortic endothelial cells (BAECs) was applied to tumor cell lines (A549 and PANC-1) and epithelial cell line (NMuMg). Cadherin conversion, expressions of α-smooth muscle actin and ZO-1, actin fiber formation and cell migration were examined as hallmarks of the induction of EMT in these cell lines. Transforming growth factor β (TGFβ) antibodies were used to neutralize TGFβ 1 , TGFβ 2 and TGFβ 3 . Expression and release of TGFβ proteins in BAECs as well as in porcine and human endothelial cells were assessed by Western blotting and ELISA, respectively. Results Conditioned medium of BAEC induced EMT in the examined cell lines. All endothelial cells from various species and locations expressed TGFβ 1 and TGFβ 2 proteins and much lower level of TGFβ 3 protein. Conditioned medium from these endothelial cells contained TGFβ 1 and TGFβ 2 , but TGFβ 3 could not be detected. Neutralizing antibody against each of TGFβ 1 or TGFβ 2 did not reverse endothelium-dependent EMT, but simultaneous neutralization of both TGFβ 1 and TGFβ 2 completely abolished it. Conclusions Endothelial cells may play a role in the induction and maintenance of EMT in tumor cells by constitutively releasing TGFβ 1 and TGFβ 2 . General significance The present results provide a novel strategy of the inhibition of tumor metastasis by targeting vascular endothelium.