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. 2019 Apr 18;9:298. doi: 10.3389/fonc.2019.00298

Figure 5.

Figure 5

Effects of miR-214 on the PI3K/Akt pathway. (A,C) The expression of PHLDA2 protein in U2OS cells after reintroduction of PHLDA2-expressing vector lacking 3′UTR (pcDNA PHLDA2). Overexpression of PHLDA2 significantly increased the radiosensitivity. (B,D) PHLDA2 vector and miR-214 mimics was co-transfected into MG63 cells. The overexpression of PHLDA2 was confirmed by western blot, which significantly antagonized the miR-214-induced radioresistance. (E) Western blot analysis for pAkt-Ser473, Akt, and GAPDH (for normalization) in miR-214-depleted U2OS cells without and with irradiation. (F) Western blot analysis of pAkt-Ser473, Akt and GAPDH (for normalization) in MG63 cells. The activation of miR-214 on Akt phosphorylation was dramatically abolished by PHLDA2 vector. (G,H) Overexpression of PHLDA2 and treatment with a PIK3CA inhibitor LY294002 significantly abrogated the inhibitory effect of miR-214 on IR-induced apoptosis. Data are presented as mean ± SD of three independent experiments. *P < 0.05.