Integrated Network Pharmacology and Experimental Analyses of the Anti-Colorectal Cancer Effects of Cinobufotalin

Objective To investigate the potential mechanism of cinobufotalin against colorectal cancer (CRC) through network pharmacology and experimental approaches. Methods Overlapping targets between cinobufotalin and CRC were identified, yielding 165 potential therapeutic targets. A protein–protein interaction (PPI) network was constructed using Cytoscape software, and key genes were subjected to Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. In vitro experiments were performed to evaluate the effects of cinobufotalin on CRC cell proliferation, apoptosis, and autophagy-related markers. Pharmacological inhibition of autophagy was also applied to examine its modulation of the antiproliferative effect of cinobufotalin. Results Enrichment analysis indicated that the overlapping targets were significantly associated with apoptosis signaling and autophagy pathways. In vitro experiments confirmed that cinobufotalin inhibited CRC cell proliferation and induced apoptosis, accompanied by changes in autophagy-related markers. Pharmacological inhibition of autophagy enhanced the antiproliferative effect of cinobufotalin, suggesting a regulatory role of autophagy in cellular response. Conclusion Cinobufotalin exerts antitumor effects against CRC by promoting apoptosis and targeting autophagy pathways, and modulation of autophagy can further enhance its therapeutic efficacy.