Fig. 1: Glutamine deprivation promotes EMT and invasion in PANC-1 cells.

A–C Gln-deficient condition promoted PANC-1 migration and invasion, and supplementation of Gln restored these phenotypes. A The cell invasion and migration ability of parental (Prt) cells and -QQ PANC-1 cells supplemented with or without two mM Gln were evaluated using Trans-well analysis. Scale bar, 200 μm. B Quantitative results of the relative migrated cell numbers were measured in (A). C Quantitative results of the relative invaded cell numbers were measured in (A). D–F Gln deficiency induced EMT in PANC-1 cells. D EMT-related transcription factors were upregulated in -QQ cells. Quantitative results of relative mRNA levels of ZEB1, ZEB2, SNAI1, SNAI2, and TWIST1 in Prt or -QQ PANC-1 cells. E Epithelial markers were downregulated, and mesenchymal markers were upregulated in -QQ cells. Quantitative results of relative mRNA levels of ZO-1, E-cadherin (E-cad, CDH1), N-cadherin (N-cad, CDH2), and vimentin (VIM) in Prt or -QQ PANC-1 cells. F E-cadherin (E-cad), N-cadherin (N-cad), and vimentin (VIM) were upregulated -QQ cells. Extracts of Prt or -QQ PANC-1 cells were analyzed by western blot assay with antibodies against E-cad, N-cad, VIM, and Ku70 (loading control). G–I Upregulated MMP2 was observed in -QQ cells. G MMP2 but not MMP9 mRNA levels increased in -QQ PANC-1 cells. Quantitative results of relative mRNA levels of MMP2 and MMP9 in the Prt or -QQ PANC-1 cells. H MMP2 protein levels increased in -QQ cells. Extracts of Prt or -QQ PANC-1 cells were analyzed with western blot assay with antibodies against MMP2, MMP9, and actin. I MMP2 activity increased in -QQ cells. MMP activities were analyzed with gelatin zymography in Prt and -QQ PANC-1 cells. Pro: pro-MMP2, active: activated MMP2. Data are represented as the mean ± SD of three independent experiments. n.s. no significance, *P < 0.05, **P < 0.01, ***P < 0.001.