Figure 6

Expression of miR-299-3p repressed activation of the PI3K/Akt pathway and downstream effector of Slug. Immunoblot analysis of phospho-Akt and phospho-PRAS40 (Akt substrate) expression and qRT-PCR of TGFB3 mRNA were performed in C4-2B and PC-3 cells overexpressing miR-299-3p cells. (A,B) Representative images of the immunoblots of Akt, pAkt, PRAS40 and pPRAS40 in the lysates of induced PC-3-299 and scr RNA control PC-3 cells. Antibodies against α-tubulin was used the internal control. (C) Comparative analysis of Akt and PRAS40 expression showing significant reduction in expression of the phosphorylated Akt and PRAS40 in PC-3-299 cells expressing miR-299-3p. Values were normalized with the internal control. Data show the mean ± SD of at least three independent assays. (D) Quantitative RT-PCR showing decreased TGFB3 mRNA expression in induced C4-2B-299 and PC-3-299 cells expressing miR-299-3p compared to uninduced or control PC-3 cells. Data show the mean ± SD of at least three independent assays. (E) A model showing miR-299-3p mediated regulation of AR and VEGFA and inactivation of downstream effectors Slug and PI3K/Akt leading to inhibition of migration and PCa progression.