miR-152 regulates the proliferation and differentiation of C2C12 myoblasts by targeting E2F3
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The development of skeletal muscle is a complex process involving the proliferation, differentiation, apoptosis, and changing of muscle fiber types in myoblasts. Many reports have described the involvement of microRNAs in the myogenesis of myoblasts. In this study, we found that the expression of miR-152 was gradually down-regulated during myoblast proliferation, but gradually up-regulated during the differentiation of myoblasts. Transfection with miR-152 mimics restrained cell proliferation and decreased the expression levels of cyclin E, CDK4, and cyclin D1, but promoted myotube formation and significantly increased the mRNA expression levels of MyHC, MyoD, MRF4, and MyoG in C2C12 myoblasts. However, treatment with miR-152 inhibitors promoted cell proliferation and restrained differentiation. Moreover, over-expression of miR-152 significantly decreased E2F3 production in C2C12 myoblasts. A luciferase assay confirmed that miR-152 could bind to the 3′ UTR of E2F3. In conclusion, this study showed that miR-152 inhibited proliferation and promoted myoblast differentiation by targeting E2F3.
KeywordsmiRNA-152 E2F3 Myogenesis C2C12 myoblasts
siRNA E2F transcription factor 3
Quantitative real-time PCR
Mailin Gan, Jingjing Du, Linyuan Shen, Dongli Yang, and Li Zhu conceived and designed the experiments. Mailin Gan and Jingjing Du performed the experiments. Mailin Gan, Anan Jiang, Qiang Li, and Yanzhi Jiang analyzed the data. Guoqing Tang, Mingzhou Li, Jinyong Wang, Xuewei Li, Shunhua Zhang, and Li Zhu contributed reagents/materials/analysis tools. Mailin Gan and Jingjing Du wrote the paper.
The study was supported by the Chinese National Sci & Tech Support Program (No. 2015BAD03B01-11), the Sichuan Sci & Tech Support Program (No. 16ZC2838, No. 16ZB0038, No. 2016NZ0089), and the earmarked fund for China Agriculture Research System (No. CARS-36-05B).
Compliance with ethical standards
The study has been performed in accordance with the approved guidelines and regulations of the Sichuan Agricultural University.
Conflict of interest
No potential conflict of interest was reported by the authors.
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