Background and Aims Ferroptosis may serve as a new target for antitumor therapy, and apoptosis inducing factor, mitochondrion-associated 2 (AIFM2) is a key regulator of ferroptosis. This study was conducted to analyze the expression level of AIFM2 in hepatocellular carcinoma (HCC), and its relationship with clinicopathologic factors and prognosis of patients, and to investigate the regulatory network of AIFM2 in HCC.Methods The differential expression of AIFM2 in HCC and normal liver tissue was analyzed using Oncomine database and the correlation between the expression of AIFM2 and the clinicopathologic characteristics of patients was determined using UALCAN database. The LinkFinder module of the LinkedOmics data platform was used for GO analysis and KEGG analysis, and the LinkedOmics database was used to analyze the protein kinases, miRNAs, and transcription factors associated with AIFM2 expression. Analysis of the protein interaction networks associated with AIFM2 was performed by the GeneMANIA database, and the frequency and type of AIFM2 mutations in HCC was defined by cBioPortal.Results The AIFM2 mRNA expression and DNA copy number in HCC tissues were higher than that in normal tissues (both P?0.05). AIMF2 was significantly associated with advanced tumors, lymph node metastasis, higher pathological grades, and TP53 mutations (all P<0.05). Patients with high expression of AIFM2 had significantly worse prognosis than those with low expression of AIFM2 (P=0.034). GO and KEGG analysis showed that AIFM2-related genes were mainly enriched in pathways related to mitochondrial and ribosomal functions, and were closely related to cellular oxidative phosphorylation and post-transcriptional translation. GSEA analysis showed that various protein kinases (MAPK1/3/7), miRNAs (miR-30 family) and transcription factors (NFAT) were significantly associated with the expression of AIFM2. C-bioportal analysis showed that the main variant form of AIFM2 in HCC was high mRNA expression but low mutation frequency.Conclusions AIFM2 is highly expressed in HCC and is associated with poor prognosis of patients. AIFM2 is a potential marker for diagnosis and prognosis of HCC. The revealed regulatory network of AIFM2 in HCC lays a foundation for the follow-up study of AIFM2 in HCC.