Indeed, qRT-PCR, immunofluorescence, and Western blotting data showed no change in the levels of PHLPP1 or PHLPP2 mRNA in miR-224-transfected cells compared to the controls (p > 0.05, Fig. 4c), whereas levels of their proteins were dramatically down-regulated in miR-224-overexpressing cells and up-regulated after inhibition of miR-224 expression in esophageal cancer cell lines (p < 0.05, Fig. 5). This evidence concerns the gene PHLPP1 and esophageal cancer.