| Summary: | The growth and development of skeletal muscle require a series of regulatory factors. MiRNA is a non-coding RNA with a length of about 22 nt, which can inhibit the expression of mRNA and plays an important role in the growth and development of muscle cells. The role of miR-22-3p in C2C12 cells and porcine skeletal muscle has been reported, but it has not been verified in Hu sheep skeletal muscle. Through qPCR, CCK-8, EdU and cell cycle studies, we found that overexpression of miR-22-3p inhibited proliferation of skeletal muscle cells (<i>p</i> < 0.01). The results of qPCR and immunofluorescence showed that overexpression of miR-22-3p promoted differentiation of skeletal muscle cells (<i>p</i> < 0.01), while the results of inhibiting the expression of miR-22-3p were the opposite. These results suggested that miR-22-3p functions in growth and development of sheep skeletal muscle cells. Bioinformatic analysis with mirDIP, miRTargets, and RNAhybrid software suggested <i>IGFBP3</i> was the target of miR-22-3p, which was confirmed by dual-luciferase reporter system assay. <i>IGFBP3</i> is highly expressed in sheep skeletal muscle cells. Overexpression of <i>IGFBP3</i> was found to promote proliferation of skeletal muscle cells indicated by qPCR, CCK-8, EdU, and cell cycle studies (<i>p</i> < 0.01). The results of qPCR and immunofluorescence experiments proved that overexpression of <i>IGFBP3</i> inhibited differentiation of skeletal muscle cells (<i>p</i> < 0.01), while the results of interfering <i>IGFBP3</i> with siRNA were the opposite. These results indicate that miR-22-3p is involved in proliferation and differentiation of skeletal muscle cells by targeting <i>IGFBP3</i>.
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