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. 2017 Dec 13;8:902. doi: 10.3389/fphar.2017.00902

FIGURE 6.

FIGURE 6

HDAC2 and HDAC5 negatively regulate the expression of miR-125a-5p in ER+ breast cancer cells. (A,B) Breast cancer cells were transfected with scramble siRNA, HDAC2 siRNA, or HDAC5 siRNA for 48 h and expression of miR-125a-5p was determined by qPCR. “∗” denotes a statistical significance (P < 0.05) between the testing groups. (C) Expression of miR-125a-5p in MCF7 and MCF7-TamC3 cells was determined by qPCR. (D) The expression of Bcl-2 and HER2 in MCF7 and MCF7-TamC3 cells were determined by Western blotting. (E) The cell surface expression HER2 (green) in MCF7 and MCF7-TamC3 cells was assessed by immunofluorescence confocal microscopy. Nuclei were countered stained by DAPI (blue). (F) Kaplan–Meier survival estimates of high (green line) or low (red line) miR-125a-5p expression in ER+ tamoxifen-treated breast cancer. (G) MCF7 and ZR-75-1 cells were transfected with either pLV-[mir-control] (empty) or pLV-[has-mir-125a-5p] (O/E miR-125a-5p) for 96 h. MCF7-TamC3 cells were pre-transfected with either pLV-[mir-control] or pLV-[has-mir-125a-5p] for 24 h and subsequently treated with or without tamoxifen for 72 h. Cell viability was determined by the MTT assay. “∗” and ∗∗∗” denote a statistical significance (P < 0.05 and P < 0.001, respectively) between the testing groups. (H) Breast cancer cells were transfected with either pLV-[mir-control] or pLV-[has-mir-125a-5p] for 48 h and expression of, HER2, Sp1, and survivin was determined by Western blotting. Actin was used as an internal control. (I) MCF7, MCF7-TamC3, and ZR-75-1 cells were transfected with either scramble or HDAC5 siRNA for 48 h and expression of survivin was determined by Western blotting.