miR-424(322)-5p targets                     Ezh1 to inhibit the proliferation and differentiation of myoblasts

Yue Yongqi; Feng Xinxin; Jia Yige; Luo Shujie; Jiang Menglei; Luo Jiandan; Hua Yonglin; Zhang Jiyue; Lin Yaqiu; Li Jian; Xiong Yan

doi:10.3724/abbs.2023036

Acta Biochimica et Biophysica Sinica (Mar 2023)

miR-424(322)-5p targets Ezh1 to inhibit the proliferation and differentiation of myoblasts

Yue Yongqi,
Feng Xinxin,
Jia Yige,
Luo Shujie,
Jiang Menglei,
Luo Jiandan,
Hua Yonglin,
Zhang Jiyue,
Lin Yaqiu,
Li Jian,
Xiong Yan

Affiliations

Yue Yongqi: ["College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, China","Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, Chengdu 610041, China"]
Feng Xinxin: ["College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, China","Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, Chengdu 610041, China"]
Jia Yige: ["College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, China"]
Luo Shujie: ["College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, China"]
Jiang Menglei: ["College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, China"]
Luo Jiandan: ["College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, China"]
Hua Yonglin: ["College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, China","Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, Chengdu 610041, China"]
Zhang Jiyue: ["College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, China","Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, Chengdu 610041, China"]
Lin Yaqiu: ["College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, China","Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, Chengdu 610041, China","Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Sichuan Province, Southwest Minzu University, Chengdu 610041, China","Key Laboratory of Animal Science of the State Ethnic Affairs Commission, Southwest Minzu University, Chengdu 610041, China"]
Li Jian: ["College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, China","Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, Chengdu 610041, China","Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Sichuan Province, Southwest Minzu University, Chengdu 610041, China","Key Laboratory of Animal Science of the State Ethnic Affairs Commission, Southwest Minzu University, Chengdu 610041, China"]
Xiong Yan: ["College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, China","Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, Chengdu 610041, China","Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Sichuan Province, Southwest Minzu University, Chengdu 610041, China","Key Laboratory of Animal Science of the State Ethnic Affairs Commission, Southwest Minzu University, Chengdu 610041, China"]

DOI: https://doi.org/10.3724/abbs.2023036
Journal volume & issue: Vol. 55
pp. 472 – 483

Abstract

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The proliferation and differentiation of myoblasts are considered the key biological processes in muscle development and muscle-related diseases, in which the miRNAs involved remain incompletely understood. Previous research reported that miR-424(322)-5p is highly expressed in mouse skeletal muscle. Therefore, C2C12 cells are used as a model to clarify the effect of miR-424(322)-5p on the proliferation and differentiation of myoblasts. The data show that miR-424(322)-5p exhibits a decreasing trend upon myogenic differentiation. Overexpression of miR-424(322)-5p inhibits the proliferation of myoblasts, manifested by downregulation of proliferation marker genes ( CCNB1, CCND2, and CDK4), decreased percentage of EdU + cells, and reduced cell viability. In contrast, these phenotypes are promoted in myoblasts treated with an inhibitor of miR-424(322)-5p. Interestingly, its gain of function inhibits the expression of myogenic regulators, including MyoD, MyoG, MyHC, and Myf5. Additionally, immunofluorescence staining of MyHC and MyoD shows that overexpression of miR-424(322)-5p reduces the number of myotubes and decreases the myotube fusion index. Consistently, inhibition of its function mediated by an inhibitor promotes the expressions of myogenic markers and myotube fusion. Mechanistically, gene prediction and dual-luciferase reporter experiments confirm that enhancer of zeste homolog 1 ( Ezh1) is one of the targets of miR-424(322)-5p. Furthermore, knockdown of Ezh1 inhibits the proliferation and differentiation of myoblasts. Compared with NC and inhibitor treatment, inhibitor+si- EZH1 treatment rescues the phenotypes of proliferation and differentiation mediated by the miR-424(322)-5p inhibitor. Taken together, these data indicate that miR-424(322)-5p targets Ezh1 to negatively regulate the proliferation and differentiation of myoblasts.

Published in Acta Biochimica et Biophysica Sinica

ISSN: 1672-9145 (Print); 1745-7270 (Online)
Publisher: China Science Publishing & Media Ltd.
Country of publisher: China
LCC subjects: Science: Chemistry: Organic chemistry: Biochemistry; Science: Biology (General): Genetics
Website: https://www.sciengine.com/ABBS/home

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