MicroRNAs play an important role in tumor cell proliferation, invasion, and Rab23 is a member of the Ras‑related small GTPase family and plays a critical role in the progression of may types of tumors. The present study was designed to investigate the inhibitory effect of microRNA (miR)‑367‑3p on the proliferation, invasion, and metastasis of prostate cancer cells. qRT‑PCR was used to detect the expression of miR‑367‑3p in prostate cancer and adjacent tissues. Cell proliferation, scratch, and Transwell assays were performed to verify the inhibitory effect of miR‑367‑3p overexpression or Ras‑related protein Rab 23 () knockdown on prostate cancer. Double luciferase reporter assay was utilized to verify whether miR‑367‑3p could target the Rab23 3'‑untranslated region (UTR). The expression levels of Rab23, Gli1, and Gli2 in prostate cancer cells transfected with the miR‑367‑3p mimic were detected via qRT‑PCR analysis. miR‑367‑3p expression in the prostate cancer tissues was downregulated compared with that in the para‑cancer control tissues. miR‑367‑3p expression in DU145 and PC3 cells was also downregulated compared with that in the human prostate epithelial cell line RWPE‑1. The overexpression of miR‑367‑3p or the knockdown of Rab23 inhibited the proliferation, invasion, and metastasis of prostate cancer cells. The results of the luciferase reporter assay confirmed that Rab23 was a target gene that was regulated by miR‑367‑3p. miR‑367‑3p specifically bound to the 3'‑UTR of Rab23 mRNA. The overexpression of miR‑367‑3p inhibited Rab23 expression and the Hedgehog pathway. Cell function experiments confirmed that the overexpression of Rab23 reversed the anticancer effect of miR‑367‑3p. miR‑367‑3p was able to inhibit the Hedgehog pathway by targeting the expression of the Rab23 gene, thus inhibiting the proliferation, invasion, and metastasis of prostate cancer cells.