Fig. 3

Changes in miR-200 s in tumor cells and plasma during breast cancer progression. a Representative hematoxylin and eosin (H&E) and FOXP3 immunohistochemical staining of breast tissues, breast tumors, and lung metastases in mice. Meta metastasis, WT wild-type, Sf scurfy, Tu breast tumor. Black arrows indicate the vasculature. b Levels of cluster 1 and cluster 2 miR-200 s (measured by quantitative (q)PCR) as percentages of snoRNA202 expression in microdissected breast epithelial cells and tumor cells. *p < 0.05 vs. WT group, two-tailed t test. Chr chromosome. c Concentrations of cluster 1 and cluster 2 miR-200 s in human breast cancer samples at various tumor stages (tumor-node-metastasis (TNM) staging classifies cancers based on T1-T4, N, and M), as determined from data for 271 patients listed in the NCI The Cancer Genome Atlas (TCGA). Data are presented as the means ± SD. *p < 0.05 vs. T1-3 group, two-tailed t test. d Plasma levels of miR-200c and miR-141 (determined by nest-qPCR) during tumor progression in Foxp3 ^sf/+ female mice. The time points of breast tumor development are indicated by vertical arrows (red arrows and blue arrows for metastatic and non-metastatic mice, respectively). Tumor progression was determined by the size and distant metastasis of tumors. The relative amounts were calculated using the 2^-∆Ct against the cycle threshold (Ct) value at 1 year of age (*p < 0.05 vs. no tumor group; two-way analysis of variance). The time points of tumor development, assessed by visual observation, are indicated by vertical arrows. All experiments were repeated three times