To evaluate whether the disruption of PD-1 affected specific CAR-dependent cytotoxic function and cytokines secretion of GPC3-CAR T cells, the in vitro tumor-lysis activity and secreted cytokines of the CRISPR-edited (or not) CAR T cells were investigated by the coculture of CAR T cells and each of various GPC3-positive (PLC/PRF/5 and SK-HEP-1/GPC3) or GPC3-negative (SK-HEP-1) HCC cells. The gene discussed is GPC3; the disease is hepatocellular carcinoma.