Background <p>To explore the biological functions, clinical significance, and regulatory mechanism of long non-coding RNA (lncRNA) ZNF667-AS1 in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC).</p> Methods <p>One hundred and seven patients with HBV-related HCC, one hundred and five patients with chronic hepatitis B (CHB) and one hundred and five healthy controls were enrolled in this study. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression levels of ZNF667-AS1, miR-210-3p, CPEB2. The association between ZNF667-AS1 expression and clinical characteristics of patients were analyzed, and receiver operating characteristic (ROC) curve analysis was performed to evaluate its diagnostic value. Bioinformatics prediction and dual-luciferase reporter assays were conducted to verify the targeted associations between ZNF667-AS1 and miR-210-3p, as well as between miR-210-3p and CPEB2. Cell Counting Kit-8 (CCK-8) and Transwell assays were applied to assess the biological functions of HCC cells after cell transfection.</p> Results <p>The expression levels of ZNF667-AS1 and CPEB2 were significantly downregulated in tissue samples from patients with HBV-related HCC and in HBV-associated HCC cell lines. ROC curve analysis showed the area under the curve (AUC) of ZNF667-AS1 for diagnosing HBV-related HCC from CHB reached 0.841. miR-210-3p was validated as a direct target of ZNF667-AS1, and a significant negative correlation was observed between their expression levels. Overexpression of ZNF667-AS1 inhibited the proliferation, migration, and invasion of HCC cells, whereas this inhibitory effect was reversed by transfection with miR-210-3p mimic.</p> Conclusions <p>LncRNA ZNF667-AS1 is downregulated in HBV-related HCC and exerts in vitro tumor‑suppressive effects by targeting miR-210-3p to upregulate CPEB2 expression, thus providing a potential auxiliary biomarker for HBV-related HCC diagnosis and a novel regulatory axis for understanding disease pathogenesis.</p>