<p>To elucidate the molecular mechanism by which long non-coding RNA CCAT1 (LncRNA CCAT1) promotes colorectal cancer (CRC) progression via regulation of microRNA miR-490-3p and Voltage-Dependent Anion Channel 1 (VDAC1), thereby activating glycolysis. In vitro, LncRNA CCAT1, miR-490-3p, and VDAC1 expression in CRC cell lines (HT-29, SW620, SW480, HCT116) and normal colonic epithelial cells (NCM460) was quantified by qRT-PCR. Selected lines underwent CCAT1, miR-490-3p, or VDAC1 overexpression/knockdown. Dual-luciferase assays confirmed interactions between CCAT1 and miR-490-3p, and between miR-490-3p and VDAC1. Cell proliferation was assessed by CCK-8 and colony formation assays; glucose consumption, ATP production, and lactate secretion were measured by ELISA. In vivo, nude mice received subcutaneous xenografts of modified HT-29 cells. Tumor growth, body weight, histopathology, apoptosis, glycolysis-related gene/protein expression (VDAC1, Hexokinase 2 [HK2], Pyruvate Kinase M2 [PKM2], Glucose Transporter 2 [GLUT2]), and glucose metabolism were evaluated. CCAT1 was significantly upregulated in CRC cells. Its knockdown suppressed proliferation and glycolysis, whereas overexpression had the opposite effect. CCAT1 acted as a competing endogenous RNA for miR-490-3p, alleviating miR-490-3p–mediated repression of VDAC1, leading to VDAC1 upregulation. Overexpressing miR-490-3p or silencing VDAC1 inhibited CRC cell proliferation and glycolysis. In vivo, CCAT1 or VDAC1 knockdown reduced tumor growth and altered glycolysis-related gene/protein expression. LncRNA CCAT1 promotes CRC progression by sponging miR-490-3p to upregulate VDAC1, thereby activating glycolysis. The CCAT1/miR-490-3p/VDAC1 axis may serve as a promising diagnostic and therapeutic target in CRC.</p> Graphical Abstract <p></p>