Yang Meng, Pang Yishan, Raza Sayed Haidar Abbas, Guo Juntao, Wang Jianfang, Zhang Gongwei, Alsahafi Sameerh, Al-Zahrani Majid, Zuo Fuyuan, Zhang Wenzhen
College of Animal Science and Technology, Southwest University, No.160 College Road, Rongchang, 402460, P.R. China.
State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, Guangzhou, Guangdong, 510275, PR China.
Mol Genet Genomics. 2025 Jun 5;300(1):56. doi: 10.1007/s00438-025-02260-8.
Myogenesis, a multistep process involving myoblast proliferation and differentiation, is critical for determining the economic value of beef cattle. While long noncoding RNAs (lncRNAs) are known to regulate myoblast proliferation, their specific mechanisms remain unclear. This study investigates the role of lncBNIP3 in bovine myoblast proliferation and examines the effects of its knockdown on cellular biological characteristics. Using quantitative real-time PCR (qRT-PCR), lncBNIP3 expression was observed to be higher in muscle tissues compared to other tissues in both 1-day-old and 24-month-old Qinchuan cattle. Knockdown of lncBNIP3 expression upregulated the mRNA levels of proliferation-related genes, as confirmed by qRT-PCR, and subsequently enhanced cellular proliferation, as demonstrated through EdU assays, flow cytometry, and CCK-8 analysis. Transcriptomic sequencing of myoblasts revealed that differentially expressed genes (DEGs) were significantly enriched in pathways associated with DNA replication and the cell cycle. Shared DEGs were primarily enriched in the minichromosome maintenance (MCM) gene family. Additionally, qRT-PCR and transcriptomic sequencing results revealed that the knockdown of lncBNIP3 expression significantly upregulated the mRNA levels of MCM family genes, including MCM2 and MCM3. Fluorescence activity assays further showed that lncBNIP3 knockdown significantly enhanced the promoter activities of MCM2 and MCM3. These findings suggest that interference with lncBNIP3 expression promotes the proliferation of bovine myoblasts, potentially through transcriptional regulation of the MCM gene family. This study provides novel insights into the regulatory functions of lncRNAs in muscle development.
成肌作用是一个涉及成肌细胞增殖和分化的多步骤过程，对于确定肉牛的经济价值至关重要。虽然已知长链非编码RNA（lncRNAs）可调节成肌细胞增殖，但其具体机制仍不清楚。本研究调查了lncBNIP3在牛成肌细胞增殖中的作用，并研究了其敲低对细胞生物学特性的影响。通过定量实时PCR（qRT-PCR）观察到，在1日龄和24月龄的秦川牛中，lncBNIP3在肌肉组织中的表达均高于其他组织。qRT-PCR证实，lncBNIP3表达的敲低上调了增殖相关基因的mRNA水平，随后通过EdU检测、流式细胞术和CCK-8分析表明细胞增殖增强。成肌细胞的转录组测序显示，差异表达基因（DEGs）显著富集于与DNA复制和细胞周期相关的通路。共享的DEGs主要富集于微小染色体维持（MCM）基因家族。此外，qRT-PCR和转录组测序结果显示，lncBNIP3表达的敲低显著上调了包括MCM2和MCM3在内的MCM家族基因的mRNA水平。荧光活性分析进一步表明，lncBNIP3敲低显著增强了MCM2和MCM3的启动子活性。这些发现表明，干扰lncBNIP3表达可能通过对MCM基因家族的转录调控促进牛成肌细胞的增殖。本研究为lncRNAs在肌肉发育中的调控功能提供了新的见解。
