Myogenesis is a complex biological process regulated by multiple factors. This study systematically revealed the dynamic changes of gene expression and its regulatory network in C2C12 myoblasts during proliferation and differentiation stages by integrating transcriptome and miRNA-omics data. The analysis results showed that in the early stage of proliferation, gene expression showed significant fluctuations, and key cell cycle and DNA replication-related genes were closely associated with specific miRNAs (miR-486a-5p, miR-486b-5p, and miR-351-5p), suggesting that these miRNAs play an important role in regulating cell proliferation. In the differentiation stage, the activation of key myogenic transcription factors and signaling pathways, such as MAPK and PI3K-Akt, synergizes with miRNA regulation to promote the myogenic program. In addition, we found that genes such as IGF1 and Dio2 were continuously upregulated during differentiation, and IGF1 might be regulated by multiple miRNAs during this process. This study provides key molecular insights for a deeper understanding of muscle development and regeneration.