Functional role of gangliotetraosylceramide in epithelial-to-mesenchymal transition process induced by hypoxia and by TGF-β

The epithelial-to-mesenchymal transition (EMT) is a basic cellular process that plays a key role in normal embryonic development and in cancer progression/metastasis. Our previous study indicated that EMT processes of mouse and human epithelial cells induced by TGF-beta display clear reduction of gangliotetraosylceramide (Gg4) and ganglioside GM2, suggesting a close association of glycosphingolipids (GSLs) with EMT. In the present study, using normal murine mammary gland (NMuMG) cells, we found that levels of Gg4 and of mRNA for the UDP-Gal: beta 1-3galactosyltransferase-4 (beta 3GalT4) gene, responsible for reduction of Gg4, were reduced in EMT induced by hypoxia (similar to 1% O-2) or CoCl2 (hypoxia mimic), similarly to that for TGF-beta-induced EMT. An increase in the Gg4 level by its exogenous addition or by transfection of the beta 3GalT4 gene inhibited the hypoxia-induced or TGF-beta-induced EMT process, including changes in epithelial cell morphology, enhanced motility, and associated changes in epithelial vs. mesenchymal molecules. We also found that Gg4 is closely associated with E-cadherin and beta-catenin. These results suggest that the beta 3GalT4 gene, responsible for Gg4 expression, is down-regulated in EMT; and Gg4 has a regulatory function in the EMT process in NMuMG cells, possibly through interaction with epithelial molecules important to maintain epithelial cell membrane organization.-Guan, F., Schaffer, L., Handa, K., and Hakomori, S. Functional role of gangliotetraosylceramide in epithelialto-mesenchymal transition process induced by hypoxia and by TGF-beta. FASEB J. 24, 4889-4903 (2010). www.fasebj.org