SLC6A14 Depletion Contributes to Amino Acid Starvation to Suppress EMT-Induced Metastasis in Gastric Cancer by Perturbing the PI3K/AKT/mTORC1 Pathway

Figure 5

SLC6A14 depletion suppresses the EMT development to delay GC metastasis by attenuating the activation of the PI3K/AKT/mTORC1 pathway. Indicated GC cells were treated with α-MT (5 mM) or LV-pTZU-SLC6A14 () plasmid for 48 h; the protein levels of N-cadherin, Vimentin, E-cadherin, and α-SMA in MGC-803 (a), BGC-823 (b), SGC7901-M (c), and MKN28-M cells (d) were checked by Western blot analysis. (e–h) GC cells were stimulated with α-MT (5 mM) or LV-pTZU-SLC6A14 plasmid (); 48 hours later, Twist1, ZEB1, ZEB2, and Slug expressions in MGC-803 (e), BGC-823 (f), SGC7901-M (g), and MKN28-M cells (h) were confirmed using Western blot assay. (i–l) Indicated GC cells were prestimulated with IGF-1 (10 ng/mL) for 16 h or were pretransfected with LV-pcDNA3.1-Raptor plasmid () for 24 h, followed by the α-MT treatment (5 mM) for another 48 h. N-cadherin, Vimentin, E-cadherin, Twist1, and ZEB2 expressions in MGC-803 (i), BGC-823 (j), SGC7901-M (k), and MKN28-M cells (l) were detected by Western blot analysis. (m–p) Indicated GC cells were pretreated with IGF-1 (10 ng/mL) for 16 h or were pretransfected with LV-pcDNA3.1-Raptor plasmid () for 24 h and then transfected with LV-pTZU-SLC6A14 () plasmid for another 48 h. N-cadherin, Vimentin, E-cadherin, ZEB2, and Twist1 expressions in MGC-803 (m), BGC-823 (n), SGC7901-M (o), and MKN28-M cells (p) were estimated by Western blot analysis. (q, r) Indicated cells were administrated with IGF-1 (10 ng/mL) for 16 h or were transfected with LV-pcDNA3.1-Raptor plasmid () for 24 h and then treated with α-MT(5 mM) (q) and LV-pTZU-SLC6A14 plasmid () (r) for another 48 h. Transwell assays were performed to identify the migration ability of GC cells. The results were demonstrated as the representative images (upper panels) and the number of migrated cells with the from three irrelevant experiments (bottom panels). . For Western blot analysis, data were demonstrated as the corresponding representatives (upper panels) and the of relative expression from triplicate experiments (bottom panels). .

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