Abstract: Objective：To determine the role of hepatocyte nuclear factor 1B (HNF1B) in the proliferation and migration of prostate cancer cells. Methods：HNF1B expression in human prostate tissues was examined by IHC, and HNF1B expression level in human prostate cells was examined by Western blotting. After overexpression of HNF1B in DU-145 cells, the cell proliferation was determined by MTS assay. Colony formation was examined by clone formation assay. Cell migration was examined by Transwell chamber assay. Results：HNF1B expressed relatively lower in tumor than that in benign tissues (P＜0.000 1). HNF1B expression in highly metastatic or hormonotherapyresistant prostate cancer cells were lower than those in lowly metastatic prostate cancer cells (P＜0.05). Overexpression of HNF1B in prostate cancer cells led to the suppression of cancer cell proliferation, migration, and colony formation (P＜0.05 or P＜0.01). Conclusions：As a tumor suppressor gene, HNF1B inhibits prostate cancer cell proliferation, colony formation, and migration.