Ethnopharmacological Relevance The medicinal relevance of Inonotus hispidus has been highlighted in various studies, particularly with respect to its antineoplastic, antioxidant, inflammation-modulating, and immunoregulatory capabilities. Nevertheless, its therapeutic value in renal cell carcinoma (RCC), especially the clear cell form (ccRCC), is yet to be defined. Aim A systematic assessment of the antitumor potential and the underlying molecular mechanisms of I . hispidus against RCC by use of network pharmacology, docking simulations, and biological experiments was undertaken as the primary objective of this research. Materials and Methods UHPLC-Q-Exactive HRMS and literature mining were used to identify bioactive constituents of I . hispidus . SwissTargetPrediction was used to predict potential molecular targets, which were then intersected with RCC-associated genes obtained in GeneCards, Therapeutic Target Database, Online Mendelian Inheritance in Man, DrugBank and PHARMGKB databases. To determine core targets, we built compound–target and protein–protein interaction (PPI) networks. GO and KEGG enrichment tools were applied for outlining the relevant pathways and biological functions to investigate the underlying biological mechanisms. Molecular docking was then implemented determine the binding potentials of active constituents with key protein targets. Antitumor activity of ethanol extract of I. hispidus (EEIH) was confirmed by in vitro experiments—CCK-8, colony formation, apoptosis, wound-healing, and Western blot—using 769-P and ACHN cell lines and in vivo xenograft models. Results 49 bioactive compounds and 169 overlapping targets of RCC were found. Based on target interactions, cerevisterol, withanolide, inonoterpene A and polyporusterone D were found to be major constituents. Network analysis and docking studies identified AKT1, EGFR, CTNNB1, STAT3, and BCL2 as core targets, exhibiting strong binding affinities with key compounds. The PI3K/Akt/mTOR and other cancer-related pathways were identified to be engaged in functional enrichment. EEIH treatment considerably inhibited RCC cell proliferation, colony formation, and migration, triggering apoptosis and downregulating phosphorylated Akt and mTOR. The in vivo antitumor efficacy of EEIH was evident in xenograft-bearing mice, where significant tumor suppression occurred in the absence of systemic toxic responses. Conclusion The antitumor mechanism of I. hispidus in ccRCC involves a network of interacting molecules and pathways, particularly those regulating PI3K/Akt/mTOR signaling. The current data support further investigation into I. hispidus as a promising natural compound for the management of ccRCC.
Wei et al. (Tue,) studied this question.