To establish patient-derived colorectal cancer organoids (CCOs) and colorectal normal organoids (CNOs), and evaluate the anti-cancer activity of toxic monomers of Traditional Chinese Medicines (TCMs).

Tissue samples were collected from patients with colorectal cancer undergoing surgical treatment in Nanjing Hospital of Chinese Medicine. CCOs and CNOs were established by 3D cell culture. The expression level of Ki-67 in tissue samples and organoids was detected with immunohistochemistry. The sensitivity of CCOs and CNOs to L-OHP, triptolide, cantharidin and bufalin was detected by CCK8 assay kits, and IC50 value was calculated.

The IC50 values of L-OHP to four CCOs and one CNO were 72.21 μM、46.78 μM、54.88 μM、128.20 μM、122.10 μM, respectively. The IC50 values of triptolide were 1.54 μM、0.19 μM、0.24 μM、0.19 μM、1.46 μM. The IC50 values of cantharidin were 34.64 μM、1.60 μM、162.61 μM、13.29 μM、146.47 μM, and that of bufalin were 0.12 μM、0.32 μM、0.43 μM、0.23 μM、0.65 μM, respectively. The four CCOs showed more sensitive to bufalin.

CCOs and CNOs were established successfully and applied in evaluation of activity of toxic components in TCMs, which provided a methodological reference.