Figure 5.

Monocytes were polarized by exosomes derived from breast cancer cells and recruited by 27-HC. A. Morphology of exosomes was determined by NP-2100 transmission electron microscope. Scale bar, 100 nm. B. HE staining of THP-1 cells was performed when cells were treated with breast cancer culture media, exosomes and exosome-depleted media. C. Expressions of CD11b and MMP-9 were detected by western blot in THP-1 cells treated with exosomes, conditional media of MCF-7 (Mm) or MDA-MB-231 (231m) cultures and PMA, respectively. D. Expressions of CD14, CD25, CD163 and IL-10 were examined by q-PCR in THP-1 cells treated with or without exosomes. GAPDH was used as the housekeeping control. E. Expression of CYP27A1 was elevated in THP-1 stimulated by exosomes derived from breast cancer cells. F. The mRNA expressions of chemokines (CCL2, CCL3 and CCL4) were examined by q-PCR in THP-1 and TDM cells (THP-1 treated with 10 nM PMA) stimulated with 1 µM 27-HC for 24 hours. G. The mRNA expressions of CCL2, CCL3, CCL4 were determined by q-PCR in peritoneal macrophages stimulated with 1 µM 27-HC for 24 hours. The values are presented as mean ± SEM (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001 vs control.