ENO1 regulates the expression of cell cycle and EMT-associated genes via FAK/PI3K/AKT pathway in NSCLC cells. (A) In A549 cells, overexpressed ENO1 increased the levels of p-Rb (ser 780) and oncogenic cell cycle regulators cyclin D1, cyclin E1, and c-Myc and decreased the expression of tumor suppressor p21. Conversely, downregulated ENO1 expression in SPCA-1 cells inhibited the expression of these proteins in addition to p21. (B) In A549 cells, overexpressing ENO1 increased the expression of EMT-marker genes including snail, vimentin, and N-cadherin and decreased the expression of E-cadherin. In SPCA-1 cells, suppressing ENO1 expression decreased the expression of these proteins in addition to E-cadherin. (C) In A549 cells, upregulated ENO1 increased levels of β-catenin, phos-FAK, PI3K, and AKT, but not their total protein levels; in SPCA-1 cells, reduced ENO1 expression reduced the levels of β-catenin, phos-FAK, PI3K, and AKT, but not their total protein levels. (D) ENO1-suppressed SPCA-1 cells were treated with Ang II (1 μmol/l) for 12 h to activate the phosphorylation of FAK, then cellular p-FAK, p-AKT, LDHA, cyclin D1, c-Myc, p21, and β-catenin were assessed by Western blot. β-Actin served as a loading control. All of the experiments were repeated at least three times.