The aim of the present study was to examine the expression and significance of peptidyl-prolyl cis-trans isomerase NIMA-interacting 1 (Pin1), β‑catenin and cyclin D1 in hepatocellular carcinoma (HCC). A total of 24 samples of HCC and adjacent normal tissues were analyzed. The expression of Pin1, β‑catenin and cyclin D1 in HCC were detected using immunohistochemistry, western blot analysis, polymerase chain reaction and immunofluorescence. The expression of Pin1, β‑catenin and cyclin D1 in HCC tissues were significantly higher than that in adjacent tissues. Pin1 was not markedly expressed in the adjacent normal tissues, while expression in the cytoplasm and nucleus of HCC cells was high. However, β‑catenin and cyclin D1 only revealed a weak expression in the cytoplasm and nucleus of HCC cells. Immunoprecipitation analyses demonstrated two clear bands at 19 and 34 kDa, and a brown band at 55 kDa as expected. Immunofluorescence analysis of HCC cells indicated that Pin1 was present in the cytoplasm and nucleus, and β‑catenin and cyclin D1 were present in the nucleus. In conclusion, the present study indicated that Pin1, β‑catenin and cyclin D1 were highly expressed in HCC. Therefore, detection of the expression of Pin1, β‑catenin and cyclin D1 may be useful for the development of novel diagnostic and treatment strategies for HCC.