〔Abstract〕 To investigate the biological functions and regulatory mechanism of hexokinase ( HK2) in driving tumor initiation and development of prostate cancer. Methods Immunohistochemical staining was employed to assess HK2 expression in benign prostate tissue and prostate cancer tissue . Correlation analyses were performed to elucidate the association of HK2 expression and clinical factors . Western blot analyses were employed to assess HK2 expression in prostate cancer cell lines . Cellular viability was assessed using the MTT assay . In vivo tumori- genesis ability of HK2 was evaluated using xenograft animal models . Glucose flux tracing was conducted through i- sotopic analyses in prostate cancer cells . Results HK2 expression was elevated in prostate cancer tissues compared to benign prostate tissues (P < 0. 05) . HK2 expression was positively correlated with Gleason Score , prostate-spe- cific antigen (PSA) levels and the presence of metastases ( P < 0. 05) . HK2 enhanced cell proliferation in vitro and tumorigenesis in vivo . HK2 augmented metabolic activity of glycolysis pathway in prostate cancer cells . Con- clusion HK2 facilitates tumorigenesis in prostate cancer by upregulating glycolytic activity .