HDAC Inhibitor MS275 Reprograms Metabolism to Induce Differentiation and Suppress Proliferation in Hepatocellular Carcinoma

Jingjie Li¹Yuyu Ye²Cheng Hu³Wen-Bo Zhu⁴Jian-Kai Liang⁴YING LIU⁴Lin Yuan⁴Liang Peng³Guang-Mei Yan⁴Ying Liu^3*

• ¹Sixth affiliated hospital of Sun Yat-sen University, Guangzhou, China
• ²Third affiliated hospital of Sun Yat-sen University, Guangzhou, China
• ³Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, China
• ⁴Department of Pharmacology, Sun Yat-sen University, Guangzhou, China

Background: Histone deacetylase (HDAC) inhibitors have shown therapeutic promise in various cancers, including hepatocellular carcinoma (HCC), due to their ability to regulate cell proliferation, differentiation, and apoptosis. However, their role in metabolic reprogramming and differentiation therapy in HCC remains underexplored. Methods: This study investigated the effects of the HDAC inhibitor MS275 on HCC cells in vitro and in vivo. Cell viability, differentiation marker expression, cell cycle distribution, metabolic activity, and reactive oxygen species (ROS) production were evaluated using CCK-8 assays, qRT-PCR, flow cytometry, Seahorse metabolic analysis, and western blotting. A xenograft mouse model was used to validate in vivo efficacy. Results: MS275 significantly suppressed HCC cell proliferation by inducing G0/G1 phase arrest without triggering apoptosis. MS275 also upregulated hepatocyte-specific markers (GLUL, HNF1A, HNF3A), indicating that it promoted differentiation. Mechanistically, MS275 reprogrammed cellular metabolism by enhancing oxidative phosphorylation and reducing glycolysis, accompanied by increased expression of the metabolic enzyme PKM1. This metabolic shift led to elevated ROS production, which was essential for MS275-induced differentiation. Knockdown of PKM1 abolished both the differentiation and anti-proliferative effects. In vivo, MS275 significantly reduced tumor growth and promoted differentiation without systemic toxicity. Conclusion: MS275 suppresses HCC cell proliferation and induces hepatocyte-like differentiation through PKM1-mediated metabolic reprogramming and ROS signaling. These findings support the potential of MS275 as a differentiation-based therapeutic strategy for HCC.