Abstract

Background & Aims This study identified zinc finger protein 544 (ZNF544) as a biomarker in hepatocellular carcinoma (HCC) and aims to delineate its functional role. Methods The correlation between ZNF544 expression and clinical parameters of HCC patients (n = 50) and the malignant biological behavior of HCC cells was analyzed in vitro (n = 5/group) and in vivo (n = 10/group). Results ZNF544 was elevated in tumors of HCC patients ( p = 0.0006). Patients with HCC exhibiting high ZNF544 expression had a higher clinical stage ( p = 0.0026) and showed greater infiltration of M2 macrophages ( p = 0.0031) and Tregs ( p = 0.0094). ZNF544 knockdown inhibited the pro-angiogenic ability of HCC cells ( p < 0.01). Increased ZNF544 expression accelerated the growth of orthotopic tumors (increased > 80%, p < 0.0001) and the infiltration of M2 macrophages (increased > 60%, p = 0.0008) and Tregs (increased > 80%, p = 0.0029) within the orthotopic tumors, and accelerated lung metastasis (number of mice with lung metastasis increased > 160%). ZNF544 bound to the spermatogenesis-associated serine-rich protein 2 (SPATS2) promoter and activated its transcription, whereas SPATS2 enhanced the triosephosphate isomerase (TPI1) mRNA stability. ZNF544 promoted glycolysis in HCC cells in a SPATS2-dependent manner. SPATS2 knockdown suppressed glycolysis in HCC cells and inhibited orthotopic tumor growth, which was rescued by TPI1 overexpression. Ivermectin was identified as a pharmacological inhibitor of ZNF544, effectively suppressing the progression of HCC. Conclusions ZNF544 accelerates HCC by activating SPATS2/TPI1 and promoting glycolysis and immune escape. Targeting ZNF544, including compounds such as ivermectin, may offer a preliminary avenue for further investigation in HCC. Impact and implications This research underscores the role of ZNF544-SPATS2 as a pivotal regulator of TPI1 expression in HCC. TPI1 shapes the immunosuppressive TME and accelerates growth and metastasi