AbstractPurposeThe underlying mechanisms of hepatocellular carcinoma (HCC) have not been fully investigated, and effective biomarkers for HCC are still needed to be explored. Therefore, our study sought to thoroughly examine the clinical significance and biological functions of the ribosomal protein L32 (RPL32) in HCC by coupling bioinformatic methods with experimental analysis.MethodsTo determine the clinical significance ofRPL32, bioinformatic analyses were performed to examineRPL32expression in HCC patient samples and to correlateRPL32expression and HCC patient survival rates, genetic alterations, and immune cell infiltration. Cell counting kit‐8 assays, colony formation, flow cytometry, and transwell assays were performed to examine the effects of RPL32 on HCC cell proliferation, apoptosis, migration, and invasion in HCC cell lines (SMMC‐7721 and SK‐HEP‐1) where RPL32 was silenced using small interfering ribonucleic acid.ResultsIn the current study, we show thatRPL32was highly expressed in HCC samples. Moreover, high levels ofRPL32were associated with unfavorable outcomes in patients with HCC. Promoter methylation and copy number variation ofRPL32were associated withRPL32mRNA expression. Results from the RPL32 silencing experiments indicated that the proliferation, apoptosis, migration, and invasion of SMMC‐7721 and SK‐HEP‐1 cells were attenuated upon RPL32 depletion.ConclusionRPL32correlates with a favorable prognosis in patients with HCC and promotes the survival, migration, and invasion of HCC cells.