This study investigates the role of mitochondrial dynamics, particularly fission, in hepatocellular carcinoma (HCC) progression. Using data from the TCGA database, researchers identified 23 mitochondrial dynamic genes and performed differential expression analysis to classify HCC patients into two clusters. Cluster 2 showed increased mitochondrial fission and worse prognosis, with upregulated fission-related genes. A ten-gene prognostic signature was developed based on differentially expressed genes between the clusters, with all genes upregulated in the high-risk group. The signature genes were found to be involved in mitochondrial fission through interaction with MTFR2, a key regulator of mitochondrial fission. High-risk patients exhibited higher TP53 mutation frequency, increased immune checkpoint expression, and lower TIDE scores, indicating worse prognosis and reduced response to immunotherapy. Immunohistochemistry and western blotting confirmed increased expression of MTFR2 and DNM1L in HCC tissues. Transmission electron microscopy showed increased mitochondrial fission in HCC tissues. The study concludes that MTFR2-related mitochondrial fission is a critical factor in HCC progression and suggests that the ten-gene prognostic signature could serve as a predictive tool and therapeutic target for HCC. The findings highlight the importance of mitochondrial dynamics in HCC and provide insights into potential therapeutic strategies.This study investigates the role of mitochondrial dynamics, particularly fission, in hepatocellular carcinoma (HCC) progression. Using data from the TCGA database, researchers identified 23 mitochondrial dynamic genes and performed differential expression analysis to classify HCC patients into two clusters. Cluster 2 showed increased mitochondrial fission and worse prognosis, with upregulated fission-related genes. A ten-gene prognostic signature was developed based on differentially expressed genes between the clusters, with all genes upregulated in the high-risk group. The signature genes were found to be involved in mitochondrial fission through interaction with MTFR2, a key regulator of mitochondrial fission. High-risk patients exhibited higher TP53 mutation frequency, increased immune checkpoint expression, and lower TIDE scores, indicating worse prognosis and reduced response to immunotherapy. Immunohistochemistry and western blotting confirmed increased expression of MTFR2 and DNM1L in HCC tissues. Transmission electron microscopy showed increased mitochondrial fission in HCC tissues. The study concludes that MTFR2-related mitochondrial fission is a critical factor in HCC progression and suggests that the ten-gene prognostic signature could serve as a predictive tool and therapeutic target for HCC. The findings highlight the importance of mitochondrial dynamics in HCC and provide insights into potential therapeutic strategies.