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. 2019 Nov 26;39(9):1983–1996. doi: 10.1038/s41388-019-1120-z

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© The Author(s) 2019

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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Fig. 7 — miR-200a inhibited Dicer expression, therefore attenuating miR-16 maturation. a Relative miR-16 stability was evaluated by real-time PCR in T24T(Vector) vs. T24T(miR-200a) cells after treatment with Act D for indicated times. b Pre-miR-16 expression was evaluated in T24T(Vector) and T24T(miR-200a) cells using real-time PCR. Bars represent means ± SD from three independent experiments. c Extracts from T24T(Vector) and T24T(miR-200a) cells were used to evaluate effect of miR-200a on Dicer expression; GAPDH used as a protein loading control. d Extracts from T24T(Vector) and T24T(KO-Dicer) cells were used to evaluate Dicer expression. GAPDH used as a protein loading control. e miRNA expression levels were evaluated by real-time PCR in T24T(Vector) vs. T24T(KO-Dicer) cells. Results were normalized to U6. f Extracts from T24T (miR-200a/Vector) and T24T(miR-200a/flag-Dicer) cells were used to evaluate Dicer expression. GAPDH used as a protein loading control. g miRNA expression levels were evaluated by real-time PCR in T24T(miR-200a/Vector) and T24T(miR-200a/flag-Dicer) cells. Results were normalized to U6. h Relative pre-miR-16 expression level were evaluated in 24T (miR-200a/Vector) and T24T(miR-200a/flag-Dicer) cells. Bars present means ± SD from three independent experiments. T24T(miR-200a/Vector) and T24T (miR-200a/flag-Dicer) cell invasion abilities were evaluated using invasion chambers (i); and the relative invasion ability was plotted (j). k Relative Dicer mRNA level was analyzed in T24T(Vector) and T24T(miR-200a) cells. l Relative Dicer mRNA stability was evaluated by real-time PCR in T24T(Vector) vs. T24T(miR-200a) cells after treatment with Act D for indicated times. m Schematic of miR-200 binding site in Dicer mRNA 3′-UTR region and its mutants aligned with miR-200a. n T24T(Vector) and T24T(miR-200a) cells were co-transfected with wild-type and mutant Dicer 3′-UTR luciferase reporters and pRL-TK, respectively. Luciferase activity of each transfectant was evaluated and results were presented as relative Dicer 3′-UTR activity. *Significant difference in Dicer 3′-UTR activity (p < 0.01). o The schematic summary of molecular mechanisms underlying miR-200a overexpression in the promotion of human BC cell invasion