SLC35A2 Deficiency Promotes an Epithelial-to-Mesenchymal Transition-like Phenotype in Madin-Darby Canine Kidney Cells.
Magdalena KotEwa MazurkiewiczMaciej WiktorWojciech WiertelakAntonina Joanna MazurAndrei RahalevichMariusz OlczakDorota Maszczak-SeneczkoPublished in: Cells (2022)
In mammalian cells, SLC35A2 delivers UDP-galactose for galactosylation reactions that take place predominantly in the Golgi lumen. Mutations in the corresponding gene cause a subtype of a congenital disorder of glycosylation (SLC35A2-CDG). Although more and more patients are diagnosed with SLC35A2-CDG, the link between defective galactosylation and disease symptoms is not fully understood. According to a number of reports, impaired glycosylation may trigger the process of epithelial-to-mesenchymal transition (EMT). We therefore examined whether the loss of SLC35A2 activity would promote EMT in a non-malignant epithelial cell line. For this purpose, we knocked out the SLC35A2 gene in Madin-Darby canine kidney (MDCK) cells. The resulting clones adopted an elongated, spindle-shaped morphology and showed impaired cell-cell adhesion. Using qPCR and western blotting, we revealed down-regulation of E-cadherin in the knockouts, while the fibronectin and vimentin levels were elevated. Moreover, the knockout cells displayed reorganization of vimentin intermediate filaments and altered subcellular distribution of a vimentin-binding protein, formiminotransferase cyclodeaminase (FTCD). Furthermore, depletion of SLC35A2 triggered Golgi compaction. Finally, the SLC35A2 knockouts displayed increased motility and invasiveness. In conclusion, SLC35A2-deficient MDCK cells showed several hallmarks of EMT. Our findings point to a novel role for SLC35A2 as a gatekeeper of the epithelial phenotype.
Keyphrases
- induced apoptosis
- cell cycle arrest
- epithelial mesenchymal transition
- endoplasmic reticulum stress
- newly diagnosed
- cell death
- signaling pathway
- emergency department
- physical activity
- genome wide
- cell proliferation
- cell therapy
- chronic kidney disease
- prognostic factors
- transcription factor
- smoking cessation
- mesenchymal stem cells
- adverse drug
- drug induced
- patient reported outcomes
- genome wide analysis