Colitis is an important risk factor for the development of colorectal cancer (CRC). The  inhibitory  effect  and  the  underlying  mechanism  of  neferine  on  colitis-associated  col-orectal cancer (CA-CRC) were investigated using an azoxymethane (AOM)/dextran sulfate sodium  (DSS)  triggered  mice  model.  Compared  with  the  CA-CRC  model,  oral  treatment  of neferine (2.5 and 5.0 mg/kg) significantly inhibited the DAI scores, decreased the tumor number, and reduced the tumor size. Neferine decreased inflammatory cell infiltration and epithelial hyperplasia in colon tissues. The levels of tumor necrosis factor-α (TNF-α), inter-leukin-1beta  (IL-1β),  and  interleukin  6  (IL-6)  in  colon  tissues  were  decreased  by  nefer-ine. Furthermore, neferine significantly decreased protein expressions of cyclooxygenase-2 (COX-2),  inducible  nitric  oxide  synthase  (iNOS),  p-p65,  and  p-STAT3  in  both  tumor  and  non-tumor tissues. In addition, neferine inhibited LPS and IL-6-induced phosphorylation of both NF-κB p65 and STAT3. Molecular docking demonstrated the interactions of neferine with both NF-κB p65 and STAT3. In conclusion, these results suggested that neferine inhib-ited CA-CRC carcinogenesis possibly by regulating NF-κB and STAT3. Neferine might be a lead compound for the chemoprevention of CA-CRC.