Fig. 4. Ectopic expression of CCND1 or PTEN attenuates miR-374a-mediated effects on proliferation, invasion, and migration in A549, pc-9, SPCA-1, and H1975 cells.

a EdU incorporation assays, b, d transwell migration assays and Boyden Chamber assays for A549, pc-9, SPCA-1, and H1975 cells were performed after transfection with NC, ectopic CCND1, PTEN, and/or miR-374a as indicated. Mean ± SD (n = 3). Scale bars, 100 μm. *P < 0.05. d miR-374a modulates expression of cell cycle-associated and EMT-related genes via inactivating PI3K/AKT and Ras pathways in A549 and pc-9 cells or activating β-catenin and Ras signalings in SPCA-1 and H1975 cells. Cell cycle regulators, including c-MYC, P21, CCND1, and CDK4, and invasion-marker genes, including ZEB2, TCF4, β-catenin, E-cadherin, and N-cadherin, were detected after transfection with NC, ectopic CCND1, PTEN, miR-374a, and/or inhibitors as indicated in NSCLC cells by western blot