Fig. 9: Circulating miR-9-5p was correlated with BC patients with metastasis.

a RT-qPCR-determined miR-9-5p levels in an equal amount of EVs derived from indicated cells (normalized to miR-16) (unpaired two-tailed t test, n = 3 independent batches). b RT-qPCR-determined miR-9-5p levels of EVs in serum from indicated xenografted mice. (one-way ANOVA, n = 3 independent batches). c Representative images of miRNA in situ hybridization (miRNA-ISH) in liver metastases from breast cancer patients. Scale bar, 50 μm. d RT-qPCR-determined miR-9-5p levels in serum from normal or BC patient with or without metastasis (unpaired two-tailed t test, n = 5). e Expression levels of miR-9-5p in BRCA tumor samples from the TCGA database. Statistical significance was assessed using two-tailed Mann–Whitney test (left, n = 1102 for Tumor and n = 104 for Normal) and paired two-tailed Student’s t test (right, n = 103). f Expression levels of miR‑9-5p in BRCA tumor samples from the GEO database. Box plots show minima, maxima, median bounds of box and quartiles (one-way ANOVA for left, n = 59 for Normal, n = 169 for Tumor, n = 54 for Metastasis; paired two-tailed t test for right, n = 59). g Expression levels of miR-9-5p in different stage of BRCA tumor samples. Box plots show minima, maxima, median bounds of box and quartiles (one-way ANOVA, n = 104 for normal, n = 182 for stage I, n = 609 for stage II, n = 244 for stage III, n = 20 for stage IV). h IHC analysis of HMGCR, CH25H, INSIG1, INSIG2 and ATF3 expression in liver metastases from breast cancer and non-BC individuals. Scale bar, 100 μm. i LC-MS/MS analysis of oxysterol derived from cholesterol in serum from normal and BC patients (unpaired two-tailed t test, n = 15 per group). j Correlation analysis of MRC1 and Ki67 expression IHC from BC patients metastatic liver. The two-tailed Pearson’s r was indicated (n = 7). k Schematic summary of this study. Data are presented as mean ± s.d. Source data and exact P value are provided as a Source data file.