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. 2019 Sep 3;28(1):313–327. doi: 10.1016/j.ymthe.2019.08.015

Figure 4.

Figure 4

miR-5188 Directly Targets FOXO1 to Augment β-Catenin-Mediated Breast Cancer Stemness, Metastasis, Proliferation, and Chemoresistance

(A) Western blot analysis of stemness, metastasis, proliferation, chemoresistance, and Wnt/β-catenin signaling-associated protein expression in miR-5188-overexpressed breast cancer cells, miR-5188-overexpressed breast cancer cells with β-catenin or miR-5188 knockdown, and their control cells. (B) Bioinformatics analysis identified the binding sequence of miR-5188 within the FOXO1 3′ UTR. (C and D) Western blot (C) and quantitative real-time PCR (D) analysis of FOXO1 expression in miR-5188-overexpressed breast cancer cells, miR-5188-silenced breast cancer cells, and their control cells. (E) Immunohistochemistry analysis of FOXO1 expression in xenograft tumors derived from MCF-7 cells after stable miR-5188 overexpression, MDA-MB-231 cells after stable miR-5188 knockdown, and their controls (scale bar, 10 μm) (n = 5). (F) Luciferase reporter assays were conducted to validate the interaction between miR-5188 and the FOXO1 3′ UTR. (G) RIP was conducted to validate the interaction between the AGO2-bound miR-5188 and FOXO1 mRNA. RIP, RNA immunoprecipitation assay. ***p < 0.001.