The invention provides a multiple immunohistochemical analysis kit for liver cancer, and a using method and application thereof, and relates to the technical field of multiple immunohistochemistry. The multiple immunohistochemical analysis kit limits that monoclonal antibody groups comprise a first monoclonal antibody group and a second monoclonal antibody group; the first monoclonal antibody group comprises a PanCK monoclonal antibody, a PDL1 monoclonal antibody, a CD8 monoclonal antibody, a CD66B monoclonal antibody and a CD163 monoclonal antibody; the second monoclonal antibody group comprises a PanCK monoclonal antibody, a PDL1 monoclonal antibody, a CD8 monoclonal antibody, a PD1 monoclonal antibody and a CD68 monoclonal antibody. According to the multiple immunohistochemical analysiskit provided by the invention, the paraffin sections of the human liver tissue in vitro are used as samples for detecting, and thus the effectiveness of the immune checkpoint inhibitor acting on theliver cancer is predicted.