MicroRNA-381 Regulates Proliferation and Differentiation of Caprine Skeletal Muscle Satellite Cells by Targeting <i>PTEN</i> and <i>JAG2</i>
In our previous study, microRNA (miR)-381 was found to be the most down-regulated miRNA in skeletal muscle of Liaoning cashmere goats with higher skeletal muscle mass, but the molecular mechanism involved remains unclear. In this study, primary caprine skeletal muscle satellite cells (SMSCs) were is...
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MDPI AG
2022-11-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/21/13587 |
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| author | Jiyuan Shen Jiqing Wang Huimin Zhen Yan Liu Lu Li Yuzhu Luo Jiang Hu Xiu Liu Shaobin Li Zhiyun Hao Mingna Li Zhidong Zhao |
| author_facet | Jiyuan Shen Jiqing Wang Huimin Zhen Yan Liu Lu Li Yuzhu Luo Jiang Hu Xiu Liu Shaobin Li Zhiyun Hao Mingna Li Zhidong Zhao |
| author_sort | Jiyuan Shen |
| collection | DOAJ |
| description | In our previous study, microRNA (miR)-381 was found to be the most down-regulated miRNA in skeletal muscle of Liaoning cashmere goats with higher skeletal muscle mass, but the molecular mechanism involved remains unclear. In this study, primary caprine skeletal muscle satellite cells (SMSCs) were isolated and identified. We investigated the effect of miR-381 on the viability, proliferation and differentiation of caprine SMSCs, and the target relationships of miR-381 with jagged canonical Notch ligand 2 (<i>JAG2</i>) and phosphatase and tensin homolog (<i>PTEN</i>). Cells isolated were positive for SMSC-specific marker protein Pax7. This suggests that purified SMSCs were obtained. The expression level of miR-381 achieved a peak value on day 4 after SMSC differentiation, and miR-381 also significantly increased the expression levels of myogenic differentiation marker genes: myosin heavy chain (<i>MyHC</i>), myogenin (<i>MyoG</i>) and myocyte enhancer factor 2C (<i>MEF2C</i>) in differentiated SMSCs, the area of MyHC-positive myotubes and the myogenic index. These findings suggest that miR-381 promoted myogenic differentiation of caprine SMSCs. The CCK8 assay and EDU staining analysis showed that miR-381 mimic both inhibited the viability of SMSCs and decreased the percentage of EDU-labeled positive SMSCs. In contrast, miR-381 inhibitor had the opposite effect with miR-381 mimic. A dual luciferase reporter assay verified that miR-381 can target <i>JAG2</i> and <i>PTEN</i> by binding to the 3′-untranslated regions (3′-UTR) of the genes. The transfection of miR-381 mimic into caprine SMSCs resulted in decreases in expression levels of <i>JAG2</i> and <i>PTEN</i>, while miR-381 inhibitor increased the two target genes in expression. This is the first study to reveal the biological mechanisms by which miR-381 regulates caprine SMSC activities. |
| first_indexed | 2024-03-09T18:59:47Z |
| format | Article |
| id | doaj.art-9e11b420f4be4e078c90dbe4b5c00c36 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T18:59:47Z |
| publishDate | 2022-11-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-9e11b420f4be4e078c90dbe4b5c00c362023-11-24T05:10:26ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-11-0123211358710.3390/ijms232113587MicroRNA-381 Regulates Proliferation and Differentiation of Caprine Skeletal Muscle Satellite Cells by Targeting <i>PTEN</i> and <i>JAG2</i>Jiyuan Shen0Jiqing Wang1Huimin Zhen2Yan Liu3Lu Li4Yuzhu Luo5Jiang Hu6Xiu Liu7Shaobin Li8Zhiyun Hao9Mingna Li10Zhidong Zhao11Gansu Key Laboratory of Herbivorous Animal Biotechnology, College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaGansu Key Laboratory of Herbivorous Animal Biotechnology, College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaGansu Key Laboratory of Herbivorous Animal Biotechnology, College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaGansu Key Laboratory of Herbivorous Animal Biotechnology, College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaGansu Key Laboratory of Herbivorous Animal Biotechnology, College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaGansu Key Laboratory of Herbivorous Animal Biotechnology, College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaGansu Key Laboratory of Herbivorous Animal Biotechnology, College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaGansu Key Laboratory of Herbivorous Animal Biotechnology, College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaGansu Key Laboratory of Herbivorous Animal Biotechnology, College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaGansu Key Laboratory of Herbivorous Animal Biotechnology, College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaGansu Key Laboratory of Herbivorous Animal Biotechnology, College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaGansu Key Laboratory of Herbivorous Animal Biotechnology, College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaIn our previous study, microRNA (miR)-381 was found to be the most down-regulated miRNA in skeletal muscle of Liaoning cashmere goats with higher skeletal muscle mass, but the molecular mechanism involved remains unclear. In this study, primary caprine skeletal muscle satellite cells (SMSCs) were isolated and identified. We investigated the effect of miR-381 on the viability, proliferation and differentiation of caprine SMSCs, and the target relationships of miR-381 with jagged canonical Notch ligand 2 (<i>JAG2</i>) and phosphatase and tensin homolog (<i>PTEN</i>). Cells isolated were positive for SMSC-specific marker protein Pax7. This suggests that purified SMSCs were obtained. The expression level of miR-381 achieved a peak value on day 4 after SMSC differentiation, and miR-381 also significantly increased the expression levels of myogenic differentiation marker genes: myosin heavy chain (<i>MyHC</i>), myogenin (<i>MyoG</i>) and myocyte enhancer factor 2C (<i>MEF2C</i>) in differentiated SMSCs, the area of MyHC-positive myotubes and the myogenic index. These findings suggest that miR-381 promoted myogenic differentiation of caprine SMSCs. The CCK8 assay and EDU staining analysis showed that miR-381 mimic both inhibited the viability of SMSCs and decreased the percentage of EDU-labeled positive SMSCs. In contrast, miR-381 inhibitor had the opposite effect with miR-381 mimic. A dual luciferase reporter assay verified that miR-381 can target <i>JAG2</i> and <i>PTEN</i> by binding to the 3′-untranslated regions (3′-UTR) of the genes. The transfection of miR-381 mimic into caprine SMSCs resulted in decreases in expression levels of <i>JAG2</i> and <i>PTEN</i>, while miR-381 inhibitor increased the two target genes in expression. This is the first study to reveal the biological mechanisms by which miR-381 regulates caprine SMSC activities.https://www.mdpi.com/1422-0067/23/21/13587miR-381caprine skeletal muscle satellite cellsviabilityproliferationdifferentiation |
| spellingShingle | Jiyuan Shen Jiqing Wang Huimin Zhen Yan Liu Lu Li Yuzhu Luo Jiang Hu Xiu Liu Shaobin Li Zhiyun Hao Mingna Li Zhidong Zhao MicroRNA-381 Regulates Proliferation and Differentiation of Caprine Skeletal Muscle Satellite Cells by Targeting <i>PTEN</i> and <i>JAG2</i> International Journal of Molecular Sciences miR-381 caprine skeletal muscle satellite cells viability proliferation differentiation |
| title | MicroRNA-381 Regulates Proliferation and Differentiation of Caprine Skeletal Muscle Satellite Cells by Targeting <i>PTEN</i> and <i>JAG2</i> |
| title_full | MicroRNA-381 Regulates Proliferation and Differentiation of Caprine Skeletal Muscle Satellite Cells by Targeting <i>PTEN</i> and <i>JAG2</i> |
| title_fullStr | MicroRNA-381 Regulates Proliferation and Differentiation of Caprine Skeletal Muscle Satellite Cells by Targeting <i>PTEN</i> and <i>JAG2</i> |
| title_full_unstemmed | MicroRNA-381 Regulates Proliferation and Differentiation of Caprine Skeletal Muscle Satellite Cells by Targeting <i>PTEN</i> and <i>JAG2</i> |
| title_short | MicroRNA-381 Regulates Proliferation and Differentiation of Caprine Skeletal Muscle Satellite Cells by Targeting <i>PTEN</i> and <i>JAG2</i> |
| title_sort | microrna 381 regulates proliferation and differentiation of caprine skeletal muscle satellite cells by targeting i pten i and i jag2 i |
| topic | miR-381 caprine skeletal muscle satellite cells viability proliferation differentiation |
| url | https://www.mdpi.com/1422-0067/23/21/13587 |
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