Research Paper Volume 13, Issue 12 pp 16341—16352

The local anesthetic ropivacaine suppresses progression of breast cancer by regulating miR-27b-3p/YAP axis

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Figure 4. MiR-27b-3p targets YAP in breast cancer cells. (A) The interaction of miR-27b-3p and YAP 3’ UTR was identified by bioinformatic analysis using Targetscan (http://www.targetscan.org/vert_72/). (BD) The MCF-7 and MDA-MB-231 cells were treated with control mimic or miR-27b-3p mimic. (B) The expression of miR-27b-3p was tested by qPCR assays in the cells. (C) The luciferase activities of wild type YAP 3’ UTR (YAP WT) and YAP 3’ UTR with the miR-27b-3p-binding site mutant (YAP MUT) were determined by luciferase reporter gene assays in the cells. (D) The mRNA expression of YAP was measured by qPCR assays in the cells. (E) The MCF-7 and MDA-MB-231 cells were treated with miR-27b-3p inhibitor or miR-27b-3p mimic. The protein expression of YAP was analyzed by Western blot analysis in the cells. (F) The MCF-7 and MDA-MB-231 cells were treated with ropivacaine (1 mmol/L) or equal volume saline, or co-treated with ropivacaine (1 mmol/L) and miR-27b-3p inhibitor. The protein expression of YAP was measured by Western blot analysis in the cells. (G) The expression of miR-27b-3p and YAP was detected in the tumor tissues from breast cancer patients (n = 50). (H) The cell viability was analyzed by MTT assays in the indicated cells. (I) The cell apoptosis was measure by flow cytometry analysis in the indicated cells. N = 3, The independent experiments were repeated for three times. Data are presented as mean ± SD. Statistic significant differences were indicated: **P < 0.01.