Conclusions
In summary, our work is the first report to reveal that miR-21 promotes hypopharyngeal cancer induced transition of bone marrow mesenchymal stem cells to cancer-associated fibroblasts, and also has an important effect on cell cycle.
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Fig. 1 Up-regulation of miR-21 in hypopharyngeal cancer and FaDu cells. a. The expression level of miR-21 in HC (hypopharyngeal cancer) and control (normal tissue). b. The expression level of miR-21 in FaDu cells and control (293T cell). All the data are presented as mean ± SD (Student’s t-test). **p < 0.01.
Fig. 2 Isolation and identification of exosomes from hypopharyngeal cancer. a. Particle size distribution curve of diluted exosomes from FaDu cells. b. Exosomes shot by electron microscope.
Fig. 3 Exosomes from FaDu cells promoted CAFs-like features in hBMSCs. a. Microscopic observation results showed the growth state of CAF-like cells in FaDu-exo+BMSC group and control. Left :293T-exo+BMSC. Right: Fadu-exo+BMSC. b. The expression level of α-SMA, FAP and GAPDH in hBMSCs on different days were analyzed by western blot. The GAPDH was used as a control. c-d. The α-SMA and FAP expression level in hBMSCs were analyzed by qRT-PCR on different days. e-g. Flow cytometry results showed the cell proportion of G1, S and G2 phase in FaDu-exo+BMSC group and control. All experiments were performed in triplicate and presented as mean ± SD. *p < 0.05, **p < 0.01. ***p < 0.001. ****p < 0.0001.
Fig. 4 Exosomes with knock-out of miR-21 inhibited CAFs-like features in hBMSCs. a. The expression level of miR-21 in exosomes or FaDu cells with miR-21 inhibited or uninhibited were analyzed by qRT-PCR. b-d. The α-SMA, FAP and GAPDH expression level of hBMSCs in miR-21 inhibitor-treated group and control were analyzed by western blot (b) and qRT-PCR (c-d) on different days. The GAPDH was used as a control. e. Microscopic observation results showed the growth state of CAF-like cells in miR-21 inhibitor treated group and control. Left: Fadu-Exo+hBMSCs. Right: miR-21-inhibitor-Fadu-Exo+hBMSCs. f-h. Flow cytometry results showed the cell proportion of G1, S, and G2 phase in miR-21 inhibitor-treated group and control. All experiments were performed in triplicate and presented as mean ± SD. *p < 0.05, **p < 0.01. ***p < 0.001. ****p < 0.0001.
Table.1 Particle size distribution of exosomes from hypopharyngeal cancer