Fig. 6: miR-205-5p LNA inhibitor reduces BCSCs metastasis in vitro and in vivo regulating EMT.
From: MiR-205-5p inhibition by locked nucleic acids impairs metastatic potential of breast cancer cells
a Representative images of transwell invasion assay of BCSCs#3 after 24-h treatment with 40 pmol LNA anti-miR-205-5p or a scramble control. Quantitative data (bottom) are presented as mean ± SD of percentage of invasion of two independent experiments each conducted in triplicate, analyzed with T-test. b Western blot analysis of Vimentin and Twist protein levels of BCSCs#3 after 24 h treatment with 40 pmol LNA anti-miR-205-5p or a scramble control; Actin was used as a loading control (upper panel). qRT-PCR of Twist, and Vimentin mRNA expression levels of BCSCs#3 treated as described above (lower panel); data are presented as mean ± SD (T-test analysis) of three independent experiments. c Representative H&E staining (upper) of lung metastasis of NSG mice (four per group) after tail vein injection with 2 × 105 BCSC#3 and treated with 20 mg/kg LNA-i-miR-205-5p or with LNA scrambled control twice a week for 2 weeks (scale bar: 400 μm). Mice were sacrificed after 30 days. Graphs show the quantification of the number (bottom left) and the size (bottom right) of lung metastasis analyzed with T-test. d Representative immunohistochemical analysis of Ki67 expression of lung metastases of NSG mice treated as described above (scale bar: 200 μm). Data are presented as percentage of Ki67-positive cells in both mice groups and analyzed with T-test (bottom)
