Potential Mechanisms of Osthole against Bladder Cancer Based on Network Pharmacology, Molecular Docking, and Experimental Validation

Abstract Backgroud: Osthole, a compound extracted from the fruit of Fructus cnidii was traditionally used in treatment for various diseases, such as male impotence, skin related dieases, and foul disease. In addition, it was reported that osthole could also inhibit various cancer. However, few studies had demonstrated that osthole could suppress bladder cancer and its mechanism was unclear. Therefore, we performed a research to explore the potential mechanism for osthole against bladder cancer. Methods: Internet web servers SwissTargetPrediction, PharmMapper, SuperPRED, and TargetNet were used to predict the osthole targets. GeneCards and the OMIM database were used to indicate bladder cancer targets. The intersection of two target gene fragments was used to obtain the key target genes. Protein–protein interaction (PPI) analysis was performed using the Search Tool for the Retrieval of Interacting Genes (STRING) database. Furthermore, we used gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses to explore the molecular function of target genes. AutoDock software was then used to perform molecular docking of target genes and osthole. Finally, an in vitro experiment was conducted to validate bladder cancer inhibition by osthole. Results: Our analysis identified 369 intersection genes for osthole, the top ten target genes included MAPK1, AKT1, SRC, HRAS, HASP90AA1, PIK3R1, PTPN11, MAPK14, CREBBP, and RXRA. The GO and KEGG pathway enrichment results revealed that the PI3K-AKT pathway was closely correlated with osthole against bladder cancer. The osthole inhibited the proliferation and migration of bladder cancer cells. Additionally, osthole blocked the bladder cancer epithelial-mesenchymal transition and promoted bladder cancer cell apoptosis by inhibiting the PI3K-AKT and Janus kinase/signal transducer and activator of transcription (JAK/STAT3) pathways. Conclusions: Osthole targets and related pathways have potential in the treatment of bladder cancer. Osthole therapy will likely inhibit proliferation and migration of bladder cancer cells by inhibiting PI3K-AKT and JAK/STAT3 pathways. Subjects: Bioinformatics, Computational Biology, Molecular Biology