Objective <p>This research explored the involvement of MAFG-AS1 in metabolic reprogramming and potential molecular mechanisms in ovarian cancer (OC).</p> Methods <p>The ability of MAFG-AS1 silencing to affect the glucose intake, lactate production, ECAR, OCR and ATP concentrations and NAD⁺/NADH ratios in OC cells was examined. Cell cycle phases and apoptosis were measured by flow cytometry. The influences of MAFG-AS1overexprssion on the above assays were also identified.</p> Results <p>A transient reduction in the number of SKOV3 and HO8910 cells in the MAFG-AS1 knockdown group. MAFG-AS1 knockdown can inhibit cell proliferation, induce apoptosis, and enhance the number of cells in G2 phase. Silencing MAFG-AS1 can inhibit the glucose uptake rate, extracellular lactate production, and ECAR of OC cells, ATP levels, and can promote OCR and NAD⁺/ NADH ratio oxidative phosphorylation. Silencing MAFG-AS1 can inhibit HIF-1α in OC.</p> Conclusion <p>Our study revealed silencing MAFG-AS1 could inhibit the proliferation and induce apoptosis of OC cells by inhibiting the HIF-1α-mediated glycolysis process. Therefore, this study further potentially reveals the machinery of MAFG-AS1 in regulating OC cell proliferation and apoptosis, which is expected to provide a theoretical basis for the study of new targets.</p>