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. 2020 Mar 20;10:5167. doi: 10.1038/s41598-020-62038-3

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Endogenous miR-299-3p expression in PCa cell lines and tissues and miR-299-3p overexpression decreased cell proliferation. (A) Average fold change in expression of miR-299-3p prostate tumor tissues (n = 15) compared to matched uninvolved areas (15), and 3 additional tumor tissues. (B) TCGA database analysis showing significant loss of expression of miR-299-3p in prostate tumors compared to normal tissues. (C) Quantitative RT-PCR showing relative fold change in miR-299-3p expression in PCa cell lines compared to non-tumorigenic RWPE-1 cells. Raw data have been normalized to the mean of RNU43, U6 and U1 snRNA. (D,E) Cell proliferation assays showing significantly reduced cell growth in miR-299-3p overexpressing cells. Data represent mean ± standard deviation (SD) of at least three independent assays in triplicates. C4-2B and 22Rv-1 cells stably transfected (D) and PC-3 cells transiently transfected (E) with inducible DNA constructs for miR-299-3p precursor miRNA or scrambled (Scr) RNA (PC-3) were induced (PC-3 at 24 h post transfection) and cell proliferation at 48 hr were detected by MTS assays. (F) Analysis of Ki67⁺ cells upon immunofluorescence staining for Ki67 performed at 72 hr post induction showing significant reduction in Ki67⁺ C4-2B and 22Rv-1 cells overexpressing miR-299-3p.