MiR-22-3p Inhibits Proliferation and Promotes Differentiation of Skeletal Muscle Cells by Targeting IGFBP3 in Hu Sheep
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 p...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-01-01
|
| Series: | Animals |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-2615/12/1/114 |
| _version_ | 1797499728470999040 |
|---|---|
| author | Shan Wang Xiukai Cao Ling Ge Yifei Gu Xiaoyang Lv Tesfaye Getachew Joram M. Mwacharo Aynalem Haile Wei Sun |
| author_facet | Shan Wang Xiukai Cao Ling Ge Yifei Gu Xiaoyang Lv Tesfaye Getachew Joram M. Mwacharo Aynalem Haile Wei Sun |
| author_sort | Shan Wang |
| collection | DOAJ |
| description | 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>. |
| first_indexed | 2024-03-10T03:51:35Z |
| format | Article |
| id | doaj.art-ba30a9920c824a4e9e0fc04abecafc48 |
| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-10T03:51:35Z |
| publishDate | 2022-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Animals |
| spelling | doaj.art-ba30a9920c824a4e9e0fc04abecafc482023-11-23T11:05:56ZengMDPI AGAnimals2076-26152022-01-0112111410.3390/ani12010114MiR-22-3p Inhibits Proliferation and Promotes Differentiation of Skeletal Muscle Cells by Targeting IGFBP3 in Hu SheepShan Wang0Xiukai Cao1Ling Ge2Yifei Gu3Xiaoyang Lv4Tesfaye Getachew5Joram M. Mwacharo6Aynalem Haile7Wei Sun8College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, ChinaJoint International Research Laboratory of Agriculture, Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou 225009, ChinaCollege of Animal Science and Technology, Yangzhou University, Yangzhou 225009, ChinaCollege of Animal Science and Technology, Yangzhou University, Yangzhou 225009, ChinaJoint International Research Laboratory of Agriculture, Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou 225009, ChinaInternational Centre for Agricultural Research in the Dry Areas, Addis Ababa 999047, EthiopiaInternational Centre for Agricultural Research in the Dry Areas, Addis Ababa 999047, EthiopiaInternational Centre for Agricultural Research in the Dry Areas, Addis Ababa 999047, EthiopiaCollege of Animal Science and Technology, Yangzhou University, Yangzhou 225009, ChinaThe 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>.https://www.mdpi.com/2076-2615/12/1/114Hu sheepskeletal muscle cellsproliferationdifferentiationmiR-22-3p<i>IGFBP3</i> |
| spellingShingle | Shan Wang Xiukai Cao Ling Ge Yifei Gu Xiaoyang Lv Tesfaye Getachew Joram M. Mwacharo Aynalem Haile Wei Sun MiR-22-3p Inhibits Proliferation and Promotes Differentiation of Skeletal Muscle Cells by Targeting IGFBP3 in Hu Sheep Animals Hu sheep skeletal muscle cells proliferation differentiation miR-22-3p <i>IGFBP3</i> |
| title | MiR-22-3p Inhibits Proliferation and Promotes Differentiation of Skeletal Muscle Cells by Targeting IGFBP3 in Hu Sheep |
| title_full | MiR-22-3p Inhibits Proliferation and Promotes Differentiation of Skeletal Muscle Cells by Targeting IGFBP3 in Hu Sheep |
| title_fullStr | MiR-22-3p Inhibits Proliferation and Promotes Differentiation of Skeletal Muscle Cells by Targeting IGFBP3 in Hu Sheep |
| title_full_unstemmed | MiR-22-3p Inhibits Proliferation and Promotes Differentiation of Skeletal Muscle Cells by Targeting IGFBP3 in Hu Sheep |
| title_short | MiR-22-3p Inhibits Proliferation and Promotes Differentiation of Skeletal Muscle Cells by Targeting IGFBP3 in Hu Sheep |
| title_sort | mir 22 3p inhibits proliferation and promotes differentiation of skeletal muscle cells by targeting igfbp3 in hu sheep |
| topic | Hu sheep skeletal muscle cells proliferation differentiation miR-22-3p <i>IGFBP3</i> |
| url | https://www.mdpi.com/2076-2615/12/1/114 |
| work_keys_str_mv | AT shanwang mir223pinhibitsproliferationandpromotesdifferentiationofskeletalmusclecellsbytargetingigfbp3inhusheep AT xiukaicao mir223pinhibitsproliferationandpromotesdifferentiationofskeletalmusclecellsbytargetingigfbp3inhusheep AT lingge mir223pinhibitsproliferationandpromotesdifferentiationofskeletalmusclecellsbytargetingigfbp3inhusheep AT yifeigu mir223pinhibitsproliferationandpromotesdifferentiationofskeletalmusclecellsbytargetingigfbp3inhusheep AT xiaoyanglv mir223pinhibitsproliferationandpromotesdifferentiationofskeletalmusclecellsbytargetingigfbp3inhusheep AT tesfayegetachew mir223pinhibitsproliferationandpromotesdifferentiationofskeletalmusclecellsbytargetingigfbp3inhusheep AT jorammmwacharo mir223pinhibitsproliferationandpromotesdifferentiationofskeletalmusclecellsbytargetingigfbp3inhusheep AT aynalemhaile mir223pinhibitsproliferationandpromotesdifferentiationofskeletalmusclecellsbytargetingigfbp3inhusheep AT weisun mir223pinhibitsproliferationandpromotesdifferentiationofskeletalmusclecellsbytargetingigfbp3inhusheep |