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AIM This study is performed to analyze the role of long non-coding RNA (lncRNA) plasmacytoma variant translocation 1 (PVT1) in prostate cancer (PCa). METHODS AND MATERIALS PVT1, miR-515-5p, and high mobility group B3 (HMGB3) mRNA expressions were examined using quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC). After gain-of-function and loss-of-function models were established, the changes in cell proliferation, migration, and invasion were evaluated using cell counting kit-8 (CCK-8) assay, 5-Ethynyl-2'-deoxyuridine (EdU) assay, and Transwell experiments. Validation of the targeting relationships between PVT1 and miR-515-5p, and between miR-515-5p and HMGB3 was conducted using bioinformatics prediction, dual-luciferase reporter assay, and RNA immunoprecipitation (RIP) experiment. Moreover, the effects of PVT1 and miR-515-5p on HMGB3 protein expression were examined using Western blot. RESULTS PVT1 expression and HMGB3 expression were up-regulated in PCa tissues and cell lines while miR-515-5p expression was down-regulated. PVT1 knockdown restrained the proliferation, migration, and invasion of LNCaP and DU145 cells in vitro, and the transfection with miR-515-5p inhibitors reversed these effects. Mechanistically, PVT1 could repress the function of miR-515; HMGB3 was proved to be a target gene of miR-515-5p, and its expression could be indirectly positively modulated by PVT1. CONCLUSION PVT1 accelerates PCa progression by repress miR-515-5p's function to upregulate HMGB3 expression. This article is protected by copyright. All rights reserved.