Fig. 3: Hypoxic stroma exosomes circHIF1A promoted breast cancer cell stemness. | Cell Death Discovery

Fig. 3: Hypoxic stroma exosomes circHIF1A promoted breast cancer cell stemness.

From: Carcinoma-associated fibroblasts derived exosomes modulate breast cancer cell stemness through exonic circHIF1A by miR-580-5p in hypoxic stress

Fig. 3: Hypoxic stroma exosomes circHIF1A promoted breast cancer cell stemness.The alternative text for this image may have been generated using AI.

A CircHIF1A was also lower in the exosomes. Cells with circHIF1A down-regulation and their exosomes were isolated and the RNA for circHIF1A measurement by real-time RT-PCR. B, C Cell proliferation in MDA-MB-231 and SKBR3 cells with exosomes from CAFs with CAFs-circRNA control/N-Exo, CAFs-circRNA control/H-Exo, CAFs-circHIF1A shRNA/N-Exo, or CAFs-circHIF1A shRNA/H-Exo treatments. The survival ability was suppressed by the CCK8 assay. D Cell proliferation in MDA-MB-231 and SKBR3 cells with exosomes from CAFs-circRNA control/N-Exo, CAFs-circRNA control/H-Exo, CAFs-circHIF1A shRNA/N-Exo, or CAFs-circHIF1A shRNA/H-Exo treatments. The survival ability was assayed by colony formation. E The data analysis from (D). F Mammosphere formation in breast cancer cells. MDA-MB-231 and SKBR3 cells were exposed to exosomes from CAFs in hypoxia or normoxia conditions. Sphere numbers became more in H-exo, and downregulation of circHIF1A reduced the numbers. The photos of mammospheres were taken under a microscope. G The data were analyzed from (F). H, I The stem cell markers decreased in the presence of H-exo with circHIF1A downregulation in MDA-MB-231 and SKBR3 cells. MDA-MB-231 and SKBR3 cells were in the presence of exosomes from CAFs-circRNA control/N-Exo, CAFs-circRNA control/H-Exo, CAFs-circHIF1A shRNA/N-Exo, or CAFs-circHIF1A shRNA/H-Exo for 6 days, and then total RNA was extracted from real-time RT-PCR. J CircHIF1A regulated stem cell-associated gene expression on protein levels. MDA-MB-231 cells and western blotting. MDA-MB-231 and SKBR3 cells were in the presence of exosomes from CAFs (CAFs-circRNA control/N-Exo, CAFs-circRNA control/H-Exo, CAFs-circHIF1A shRNA/N-Exo, or CAFs-circHIF1A shRNA/H-Exo) for 6 days, and then total protein was extracted for western blot. *p < 0.05, **p < 0.01.

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