EGF up-regulates <i>miR-31</i> expression by virtue of AKT activation in OSCC cells.

<p>(A) The Graphic algorithm was used to illustrate the expression profile of 14 miRNAs following EGF treatment in SAS and HSC-3 cells. Green, up-regulated; red, down-regulated. (B–E) SAS cells. (B, D, E) qRT-PCR analysis. (C) Western blot analysis. (B) Inhibition of potential EGF downstream signals by pretreatment with LY294002 and U0126. LY294002 decreased endogenous <i>miR-31</i> expression as well as EGF induced <i>miR-31</i> expression. (C) Exogenous AKT expression and AKT activation mediated by plasmid transfection. (D) Up-regulation of <i>miR-31</i> expression induced by AKT activation. (E) LY294002 administration blocked both endogenous <i>miR-31</i> expression and AKT induced <i>miR-31</i> expression. VA, vector alone. Numbers below pictures are normalized values. Data in (A) are from duplicate experiments. Other data are the means ± SE from at least triplicate analysis. <i>ns</i>, not significant; *, <i>p</i><0.05; **, <i>p</i><0.01; ***, <i>p</i><0.001; un-paired <i>t</i>-test.