Functional Validation of ALDOA in Regulating Muscle Cell Fate: Based on In Vitro Proliferation, Apoptosis, and Differentiation Experiments

<b>Background/Objectives</b>: This study systematically investigated the expression characteristics of the <i>ALDOA</i> gene in skeletal muscle cells and its effects on cell proliferation, apoptosis, and differentiation. <b>Methods</b>: We constructed an <i>ALDOA</i> overexpression vector and transfected it into C2C12 cells and porcine skeletal muscle satellite cells. <b>Results</b>: We found that <i>ALDOA</i> exhibited the highest expression in the longissimus dorsi muscle and was primarily localized in the cell nucleus. Overexpression of <i>ALDOA</i> significantly inhibited cell proliferation, induced G0/G1 phase arrest, and downregulated the expression of proliferation-related genes such as <i>CDK2</i> and <i>Cyclin D1</i>. Concurrently, <i>ALDOA</i> overexpression markedly promoted apoptosis. Regarding differentiation, although <i>ALDOA</i> expression was upregulated during differentiation, its overexpression significantly suppressed the expression of myogenic differentiation-related genes (such as <i>MYOD</i>, <i>MYOG</i>, <i>MEF2C</i>), suggesting a negative regulatory role in differentiation control. <b>Conclusions</b>: This study reveals the multifaceted regulatory functions of <i>ALDOA</i> in skeletal muscle cells, providing experimental evidence for deepening the understanding of its mechanisms in muscle development and regeneration. This study provides the first functional evidence that <i>ALDOA</i> acts as a multifunctional regulator in skeletal muscle cells, negatively governing cell growth and fate decisions by inhibiting proliferation, promoting apoptosis, and impeding myogenic differentiation, thereby extending its role beyond glycolysis to direct governance of cellular processes. This study reveals for the first time that <i>ALDOA</i> possesses dual functions in muscle cells, regulating both metabolism and transcription.