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首页> 外文期刊>Gene: An International Journal Focusing on Gene Cloning and Gene Structure and Function >bta-miR-378 promote the differentiation of bovine skeletal muscle-derived satellite cells
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bta-miR-378 promote the differentiation of bovine skeletal muscle-derived satellite cells

机译:BTA-MIR-378促进牛骨骼肌衍生卫星细胞的分化

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摘要

The mechanism by which bta-miR-378 regulates bovine skeletal muscle-derived satellite cell (bMDSC) myogenesis remains unknown. In this study, stem-loop RT-PCR was used to assess bta-miR-378 expression during the proliferation and differentiation of bMDSCs. The results showed that bta-miR-378 expression did not obviously change during bMDSC proliferation but increased significantly when bMDSCs began to differentiate. Then, a bta-miR-378 mimic (bta-miR-378-M) and bta-miR-378 inhibitor (bta-miR-378-I) were transfected into bMDSCs to explore the effect of bta-miR-378 on bMDSC differentiation. Cell differentiation was detected using myosin heavy chain 3 immunofluorescence, myotube formation, and desmin and myogenin western blotting analyses. As expected, bta-miR-378-M enhanced bMDSC differentiation, whereas bta-miR-378-I had the opposite effect. Moreover, luciferase reporter and western blotting assays showed that bta-miR-378 directly targeted the 3'-untranslated regions of DNA polymerase alpha subunit B (POLA2) to regulate its protein expression. In summary, these data indicate that bta-miR-378 targets POLA2 to promote the differentiation of bMDSCs, which provides further insight into the biological functions of bta-miR-378 in bovines.
机译:BTA-MIR-378调节牛骨骼肌衍生卫星细胞(BMDSC)肌生成的机制仍然未知。在该研究中,茎环RT-PCR用于评估BMDSC的增殖和分化期间的BTA-miR-378表达。结果表明,BMDSC增殖期间BTA-MIR-378表达明显没有变化,但当BMDSC开始区分时,显着增加。然后,将BTA-miR-378模拟(BTA-miR-378-M)和BTA-miR-378抑制剂(BTA-miR-378-I)转染到BMDSC中,以探讨BTA-MIR-378对BMDSC的影响差异化。使用肌菌素重链3免疫荧光,肌室形成和脱霉素和肌原素Western印迹分析来检测细胞分化。正如预期的那样,BTA-MIR-378-M增强的BMDSC差异,而BTA-MIR-378-I具有相反的效果。此外,荧光素酶报告者和蛋白质印迹测定表明,BTA-miR-378直接靶向DNA聚合酶α亚基(POLA2)的3'非翻译区以调节其蛋白质表达。总之,这些数据表明BTA-MIR-378靶向POLA2以促进BMDSC的分化,这提供了进一步了解BTA-MIR-378在牛的生物学功能。

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