To explore the mechanism of jolkinolide B in the treatment of carcinoma of colon based on network pharmacology and in vivo/vitro experiment.The Target of jolkinolide B was predicted by TCMSP and other databases,and the mechanism of jolkinolide B against carcinoma of colon (CRC) was analyzed by Target enrichment analysis and molecular docking.Based on the prediction results,in vivo experiment was used to observe the inhibitory effects of jolkinolide B on the growth of transplanted tumor.The MTT assay was used to detect the proliferation inhibitory activity of jolkinolide B at different concentrations on HCT116 cells,flow cytometry was used to detect cell apoptosis,and a cell scratch assay was used to observe the effects of jolkinolide B on migration ability of HCT116 cells.Western blot was used to detect the expression of related apoptotic proteins Bax,Bcl-2,MMP3 and MMP12.In network target analysis,jolkinolide B matched with CRC to get 78 targets.Enrichment signal pathways mainly include IL-17 signal pathway,PD-1/PD-L1 signal pathway,etc.Molecular docking results showed that jolkinolide B bonded to STAT3,MAPK8 and Jun core proteins by hydrogen bonds,and the binding conformation was relatively stable.In vivo experiment showed that jolkinolide B obviously reduce the volume of the transplanted tumor.The results of cell-level verification experiments showed that jolkinolide B significantly inhibited the proliferation of HCT116 cells,promoted cell apoptosis,and inhibited cell migration;Western blot results showed that jolkinolide B significantly up-regulates the expression of Bax/Bcl-2,which induces cell apoptosis,and down-regulates the expression of MMP3 and MMP12,which inhibit tumor cell invasion and metastasis.This study provides a reference for the study on the clinical drug use and the mechanism of action of jolkinolide B.