Medulloblastoma (MB) is the most malignant paediatric brain cancer and current curative rates have not significantly improved recently. New diagnostic and prognostic biomarkers and improved treatment methods are needed to enhance MB patient outcomes. Our previous study showed that LIN28B, an RNA-binding protein, is a potential diagnostic and prognostic marker for MB, as well as a pharmacological target to inhibit the proliferation and stemness of MB cells. However, the specific role of LIN28B protein and its mechanism of action in MB had not been studied to date. This study assessed the role of LIN28B in Daoy MB cells using siRNA-mediated silencing of LIN28B. Supported by proteomics analysis and cell-based assays, silencing of LIN28B reduced the viability of MB cells, as well as ribosome biogenesis, and promotes cellular lipid accumulation. This study highlights the importance of LIN28B as a promising target to regulate MB cell growth, ribosomal biogenesis and lipid metabolism.