Objective : 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 vivotumorigenesis ability of HK2 was evaluated using xenograft animal models. Glucose flux tracing was conducted through isotopic 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-specific antigen(PSA) levels and the presence of metastases(P<0.05). Overexpression ofHK2enhanced cell proliferationin vitro(P<0.01). Knockdown ofHK2decreased tumorigenesisin vivo. HK2 augmented metabolic activity of glycolysis pathway in prostate cancer cells(P<0.05). Conclusion HK2 facilitates tumorigenesis in prostate cancer by upregulating glycolytic activity.