Proteins that bind to microtubule are important for cell cycle, and some of these proteins show oncogenic characteristics with mechanisms not fully understood Herein we demonstrate overexpression of protein regulator of cytokinesis 1 (PRC1), a microtubule associated regulator of mitosis, in human hepatocellular carcinoma (HCC) Moreover, upregulated PRC1 is associated with lower survival rates of HCC patients Mechanistically, reducing PRC1 blocks mitotic exit of HCC cells at telophase in a spindle assembly checkpoint independent manner, and acts synergistically with microtubule associated agents (MTAs) to suppress p53-wt or p53-null HCC cells in a p53-or p14ARF-dependent manner, while overexpressing PRC1 increases the resistance of HCC to taxol A combined treatment of taxol/shPRCI results in 90% suppression of tumor growth in subcutaneous HCC xenograft models In orthotopic xenograft mice, reducing PRC1 significantly alleviates HCC development and hepatic injury Together, our results suggest a dual mitotic suppression approach against HCC by combining MTAs with cytokinesis inhibition, which blocks mitosis at both metaphase and telophase.