TRIM28 Regulates Hepatocellular Carcinoma Progression and Aerobic Glycolysis through Interacting with PFKFB3

Abstract

Background: Hepatocellular carcinoma (HCC) is a common and deadly malignancy with poor clinical outcomes. Aerobic glycolysis is a hallmark of metabolic reprogramming in HCC progression. Methods: Validation of TRIM28 (tripartite motif containing 28) expression was conducted by real-time reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry assay using clinical HCC samples. Function-based experiments were carried out using TRIM28 siRNA transfection in vitro, including cell counting kit-8 (CCK-8), ethynyl-2-deoxyuridine (EDU) staining and transwell assay. Pyruvate, lactate concentration and adenosine triphosphate (ATP) level were examined through the glucose, lactate, and ATP assay kits, respectively. TRIM28 and PFKFB3 (6-phosphofructo-2-kinase) expression and colocalization were determined by immunofluorescence. RNA pulldown experiment was applied to elucidate the underlying interaction between TRIM28 and PFKFB3. Overexpression of PFKFB3 significantly reversed the TRIM28-induced cell proliferation, migration and aerobic glycolysis. Results: TRIM28 was up-regulated in both HCC clinical samples and cell lines. Silencing of TRIM28 inhibited cell proliferation, migration and aerobic glycolysis levels in Huh-7 cells. Immunofluorescence and RNA pulldown assays confirmed the interaction between TRIM28 and PFKFB3. Moreover, it was observed that overexpressing PFKFB3 could rescue the inhibitory impacts of TRIM28 siRNA on cell proliferation, migration and aerobic glycolysis. Conclusions: Our findings suggest a preclinical proof of concept for TRIM28 as a potential therapeutic target in HCC.