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Fig. 2 | Breast Cancer Research

Fig. 2

From: MicroRNA profiling of the pubertal mouse mammary gland identifies miR-184 as a candidate breast tumour suppressor gene

Fig. 2

Expression of miR-184 is lost in cancer models. Ectopic expression suppresses proliferation and self-renewal in vitro and in vivo. a miR-184 expression is lost in several mouse models of breast cancer, compared to total mouse mammary epithelium. microRNA expression was normalised to SnoRNA202. b miR-184 is undetectable in a majority of human breast cancer cell lines. microRNA expression is normalised to RNU6. c MDA-MB-231 were transfected with microRNA mimics, proliferation was measured for 5 days by MTS assay. d BT-549 were transfected with microRNA mimics, proliferation was measured for 5 days using MTS assay. c, d Results are expressed as mean + standard error from three independent experiments performed in sextuple technical replicates. The t test was performed: *p <0.05 indicates significant difference in proliferation in cells overexpressing miR-184 compared with non-targeting miRNA control. e MDA-MB-231 stably overexpressing miR-184 and control were cultured in low adherent plates and enumerated for primary and secondary tumourspheres. The t test was performed: *p <0.05 indicates significant difference in tumoursphere-forming potential in cells overexpressing miR-184 compared to control. f Tumour mass in NOD/SCID mice (n = 5) after injection of MDA-MB-231 constitutively overexpressing miR-184, let-7a or negative control into the mammary fat pad. g Haematoxylin and eosin (H&E) (top) and phospho-histone H3 (bottom) immunohistochemical analysis. Circle identifies focus of MDA-MB-231 cells at the injection site. Top scale bars = 50 μM, bottom scale bars = 30 μM. h Quantitation of phospho-histone H3 immunoreactivity and mitotic figures in tumour sections. The t test was performed: *p <0.05 indicates significant difference in positive phospho-histone H3 expression and mitotic figures in miR-184 cohort compared to control cohort. HER2 human epithelial growth factor receptor 2, neg negative

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