When HCC cells and patient‐derived organoids are cultured on 2D plastic or 3D matrix gel but in the presence of a RHO/ROCK inhibitor, the functional alterations were not observed, suggesting that ECM modification may be required for THBS2‐mediated change in ability of HCC cells to migrate, invade, self‐renew, and resist 5‐FU treatment. Here, RHO is linked to hepatocellular carcinoma.