CENPO Drives Cutaneous Squamous Cell Carcinoma Metastasis via BIRC5- Dependent Glycolytic Regulation

Abstract Background Cutaneous squamous cell carcinoma (cSCC) is the second most common skin malignancy. Metastasis is an important cause of death in cSCC patients. CENPO overexpression in cSCC. However, the relationship between CENPO and cSCC metastasis remains unclear. Methods The protein levels in clinical specimens were investigated by bioinformatics analysis and immunohistochemistry (IHC). The CENPO, BIRC5, GLUT1, HK2 and PKM2 proteins were analyzed by western blotting. Additionally, the mRNA levels of CENPO and BIRC5 were assessed using quantitative polymerase chain reaction (Q-PCR). Cell proliferation was evaluated using the celigo imaging system, while cell migration was assessed via transwell. Cell invasion was measured by wound healing assays. Glycolytic parameters, including oxygen consumption rate (OCR), extracellular acidification rate (ECAR), glucose, pyruvate, lactate, and ATP levels were quantified using a multifunctional fluorescent microplate reader. Furthermore, the mechanism of CENPO in vivo was evaluated with subcutaneous xenograft models. Results Compared with normal tissue, CENPO was significantly upregulated in cSCC tissues and correlated with poor prognosis. CENPO knockdown suppressed proliferation, migration, and invasion in vitro (P<0.001). BIRC5 was the strongest correlation with CENPO (R = 0.57). CENPO silencing downregulated BIRC5, and GLUT1/HK2/PKM2 proteins levels (P<0.001). CENPO/BIRC5 overexpression decreased OCR (P<0.001) but increased ECAR (P<0.001), glycolytic intermediates (P<0.001), and ATP production (P<0.001). Conclusion Our study identifies CENPO as a novel metabolic driver in cSCC pathogenesis. The CENPO promotes tumor metastasis through BIRC5 dependent GLUT1/HK2/PKM2-mediated aerobic glycolysis, suggesting a therapeutic targeting potential for metabolic intervention in cSCC.