Fig. 4. MiR-7641 regulated cancer stem cell properties and glycolysis.
A miR-7641 level in CAFs. The total RNA was extracted from CAFs transfected with miR-7641 and its expression was assayed by real-time RT-PCR. B miR-7641 levels in sEVs from CAFs. The total RNA was extracted from sEVs of CAFs transfected with miR-7641 (CAFs-sEV/miR-7641) or its control (CAFs-sEV/miR-con) and its expression was assayed by real-time RT-PCR. C, D miR-7641 suppressed breast cancer cell proliferation. MDA-MB-231 and SKBR3 cells were treated with CAFs-sEV/miR-con or CAFs-sEV/miR-7641 for CCK8 assay in five days. E, F Colony formation of MDA-MB-231 and SKBR3 cells. MDA-MB-231 and SKBR3 cells were treated with CAFs-sEV/miR-con or CAFs-sEV/miR-7641, and then seeded in 6-well plates, and colonies were observed and taken photos after 10 days. G, H Sphere formation rates of MDA-MB-231 and SKBR3 cells. MDA-MB-231 and SKBR3 cells were treated with CAFs-sEV/miR-con or CAFs-sEV/miR-7641, and then seeded in 6-well plates, and spheres were observed and taken photos after 7 days. I, G ECAR in MDA-MB-231 cells. Cells were treated with CAFs-sEV/miR-con or CAFs-sEV/miR-7641, and ECAR was analyzed using seahorse. K, I ECAR in SKBR3 cells. Cells were treated with CAFs-sEV or CAFs-sEV with miR-7641, and ECAR was analyzed using seahorse. M, N The expression of stem cell and glycolysis associated genes in breast cancer cells. The total RNA was extracted from breast cancer cells treated CAFs-sEV/miR-con or CAFs-sEV/miR-7641. The markers such as CD44, CD24, and ALDH1 were assayed by real-time RT-PCR. O The expression of stem cell and glycolysis associated genes in breast cancer cells. MDA-MB-231 and SKBR3 cells were treated with CAFs-sEV/miR-con or CAFs-sEV/miR-7641 for 48 h and total protein was extracted for western blotting analysis. con: control. *p < 0.05, **p < 0.01.