The invention discloses an in-vitro construction method of a liver cancer organ model. Liver cancer cells, hepatic stellate cells and liver sinusoidal endothelial cells are suspended in a culture medium A in a specific proportion; on the fourth day of culture, half of culture liquid is replaced, and culture is maintained; from the seventh day, the culture medium A is replaced with a culture mediumB, and then culture is continuously conducted for seven days; amplification subculture is conducted on the fourteenth day. On the basis of complex cellularity in a tumor microenvironment, the 3D liver cancer organ in-vitro model which is uniform in size, stable in structure, capable of being used for detecting the effectiveness of anticancer drugs and capable of achieving multiplication culture is quickly constructed. The constructed liver cancer organ model is simple in method, quick to construct, high in operability and suitable for researching a liver cancer generation and development mechanism, high-throughput screening of liver cancer drugs and the like, and has industrialization significance.