In this paper, information of the main active ingredients in grape seed oil and corresponding target proteins were obtained through literature search and TCMSP database, cancer and tumor-related targets were screened out from Genecards and OMIM databases and common targets for drugs and diseases were gained by the Venn online platform, “active ingredient-disease target” network diagram were constructed with Cytoscape 3.7.2 software, protein interaction (PPI) network were drawn by String database, GO (Gene Ontology) function enrichment and KEGG signal pathway analysis for key targets were performed by David database. Furthermore, the anti-cancer and anti-tumor functional components and mechanism of grape seed oil were further explored based on the method of network pharmacology. The results showed that a total of 15 active ingredients in grape seed oil were screened, 236 targets were obtained, 686 cancer and tumor-related targets were obtained from the disease database, and 93 common targets for drugs and diseases were obtained through the Venn diagram. PPI network analysis showed the key anti-cancer and anti-tumor targets of grape seed oil were signal transducer, transcription activator 3, mitogen-activated protein kinase 1, mitogen- activated protein kinase 3, and cell tumor antigen p53. GO functional analysis obtained 477 biological processes with P < 0.05, which were mainly involved in regulating biological functions such as enzyme binding, protein binding, transcription factor binding, protein heterodimerization activity, and DNA binding in the transcription regulatory region. KEGG pathway enrichment analysis obtained 113 pathways with P < 0.05, involving NOD-like receptor signaling pathway, MAPK signaling pathway, p53 signaling pathway, ErbB signaling pathway, mTOR signaling pathway, VEGF signaling pathway, Wnt signaling pathway and other classic signaling pathways. The research predicted the anti-cancer and anti-tumor functional components, targets, biological processes and enrichment pathways of grape seed oil, revealed its mechanism of action, and provided references for further research on the anti-cancer effects of grape seed oil.