Hepatocellular carcinoma is the most common type of primary liver cancer. Worldwide, metabolic dysfunction-associated steatotic liver disease and metabolic dysfunction-associated steatohepatitis are becoming the most common triggers of liver disease and are now major contributors to hepatocellular carcinoma development. Metabolic dysfunction-associated steatotic liver disease triggers hepatic lipid accumulation, inducing oxidative stress, inflammatory activation, and fibrosis, culminating in hepatocellular damage and proliferation, key drivers of metabolic dysfunction-associated steatohepatitis associated hepatocellular carcinoma. Translational studies in human cell lines and mouse models have identified serine/threonine kinase (STK)25 in regulating liver lipid homeostasis and the progression of metabolic dysfunction-associated steatotic liver disease/metabolic dysfunction-associated steatohepatitis. In this study, a nonclinical proof-of-principle investigation was performed to assess the benefits of Stk25- antisense oligonucleotides and N-acetylgalactosamine Stk25 antisense oligonucleotides, which were either designed to be hepatocyte-specific or broadly distributed to peripheral organs, in blocking the initiation and progression of metabolic dysfunction-associated steatohepatitis related hepatocellular carcinoma in mice. Results revealed reduced hepatic messenger RNA expression of genes controlling apoptosis, fibrosis, proliferation, oxidative/endoplasmic reticulum stress, and inflammation in mice treated with both Stk25- antisense oligonucleotides and N-acetylgalactosamine Stk25 antisense oligonucleotides, alongside decreased immunolabeling for proliferative and hepatocellular carcinoma diagnostic markers. Additionally, a 90% reduction in STK25 abundance was observed in Stk25- antisense oligonucleotides and N-acetylgalactosamine Stk25 antisense oligonucleotides-dosed mice. While STK25 antagonism effectively suppressed hepatocellular carcinoma development, it did not entirely prevent its occurrence. These findings suggest that targeting STK25 presents a promising therapeutic strategy for metabolic dysfunction-associated steatohepatitis-driven hepatocellular carcinoma.