Phospho-p70S6K inhibitor (PF-4708671) suppressed phospho-p70S6K expression and cell proliferation in SCC-4 and SCC-25 cells. (A) As determined using Western blotting, the phospho-p70S6K (p-p70S6K) inhibitor (PF-4708671) reduced the expression of phospho-mTOR (p-mTOR) and p-p70S6K in SCC-4 and SCC-25 cells. SCC-4 and SCC-25 cells were not treated as negative control (NC) or treated with 20 µM PF-4708671 for 6 h (6 h), 12 h (12 h), 18 h (18 h), 1 day (1 d), 2 days (2 d), and 3 days (3 d). Western blot analysis was performed to examine the expression levels of p-mTOR, mTOR, p-p70S6K, p70S6K, and β-actin. (B) Inhibitory effects of various doses (0.5, 1, 2, 5, 10, 20, and 40 µM) of PF-4708671 on SCC-4 (B) and SCC-25 (C) cells were assessed using MTT assay after 72 h of treatment. Data presented are the mean and standard error of the mean of three independent experiments. * p < 0.05, ** p < 0.01, and *** p < 0.001 indicate the statistical significance of differences between cells with and without treatment. t (red ** and *** for SCC-4 and blue *, ** and *** for SCC-25).