Prostate cancer is the most common cancer in men, and before it progresses and metastasizes, the anticancer drug bicalutamide is often administered to patients. It can develop into the androgen-independent form, which is a serious progressive and metastatic state. Many cases of androgen-dependent prostate cancer develop resistance during treatment with bicalutamide. Therefore, the effect of bicalutamide on androgen-dependent LNCaP prostate cancer cells is of clinical interest. The aim of this study was to demonstrate the effects of the anticancer drug bicalutamide on LNCaP prostate cancer cells by using a proteomics approach. Based on the results, we identified > 2-fold differentially expressed proteins, and 347 proteins were differentially expressed between the normal RWPE-1 prostate cells treated with androgen and LACaP cells treated with androgen. Furthermore, 314 proteins were differentially expressed between the LNCaP and LNCaP-Bic groups. The apoptosis pathway associated with differentially expressed proteins was shown in the Kyoto Encyclopedia of Gene and Genome (KEGG) pathway mapper. The KEGG pathway mapper results revealed that the fodrin-mediated apoptosis pathway is associated with the actions of bicalutamide and western blotting was performed to validate these results.