Fig. 5

PFKL promotes the proliferation and migration of oral squamous cell carcinoma cells by upregulating glucose metabolism. A Western blotting analysis was performed to confirm USP14 expression in OSCC tissues and adjacent normal tissues (n = 8). B Pearson analysis was performed to determine the correlations between USP14 and PFKL in OSCC tissues. C, D Overall survival (OS) and disease specific survival (DSS) based on PFKL expression in OSCC (TCGA). PFKL high (red) group corresponds to the fourth quartile of expression, while PFKL low (blue) group corresponds to the first quartile. E The protein levels of PFKL in four OSCC cell lines were compared with a normal oral epithelial cell line HOK by Western blotting analysis. F, G Western blotting results revealed the knockdown efficiency of PFKL in HN4 and HN6 cells. H, I HN4 and HN6 cells were transfected with pGIPZ-shPFKL plasmids. Proliferative ability was determined by CCK8 assay at the indicated time points. Data represent the means ± S.D. of three independent experiments. J, K OSCC cells were transfected with the indicated plasmids and seeded into 6-well plates at a density of 2000 cells/well. After 10 days, colony formation was detected by crystal violet staining. Data represent the means ± S.D. of three independent experiments. L, M Wound healing assay was performed on OSCC cells stably transfecting with PFKL-shRNA plasmids. Images were photographed and analyzed by measurement of the cell-free areas in multiple fields using a service provided by Wimasis. Data represent the means ± S.D. of three independent experiments. N, O OSCC cells were transfected with PFKL-shRNA plasmids. Cell migration was investigated using Transwell assay. The numbers of migrated cells per field (mean ± S.D.) from three independent experiments. P–S Cellular glucose consumption and lactate excretion were detected in OSCC cells with PFKL silencing using glucose uptake assay and lactate colorimetric assay, respectively. Data represent the means ± S.D. of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001