Therapeutic effects of the n-butanol extract of Potentilla freyniana Bornm. in hepatocellular carcinoma cells.

OBJECTIVE

This study aimed to integrate network pharmacology, bioinformatics analysis, molecular docking, and experimental validation to construct a "component-target-pathway" multidimensional network model, systematically elucidate the potential mechanisms underlying the therapeutic effects of the extract of Potentilla freyniana Bornm. (PFB) on hepatocellular carcinoma (HCC), and thereby clarify its pharmacological basis.

METHODS

HCC datasets were retrieved from GEO and TCGA databases, and the DEGs were screened. The active components of the n-butanol extract of PFB were obtained by UHPLC-MS/MS, and the candidate target genes were predicted by the SwissTargetPrediction, Similarity Ensemble Approach, and SuperPred databases. The overlapping target genes were selected by GO and KEGG enrichment analysis, and the key target genes were screened by the SVM and RF algorithms. The verification of differentially expressed target genes and ROC analysis of key target genes were performed. Molecular docking was performed using CB-Dock2. We investigated the parameters of proliferation, migration, invasion, and apoptosis in the n-butanol extract of PFB treated HCC, and we verified the expressions of key proteins in HCC by Western blot.

RESULTS

Toxicity experiments showed that the n-butanol extract of PFB did not cause significant toxic damage to the mice heart, liver, and kidney. CCK8 assays detected that the n-butanol extract of PFB had inhibitory effects on HCC. Through network pharmacology, we obtained a total of 17 overlapping genes and finally screened out 6 key target genes by SVM and RF algorithm analyses. Molecular docking and molecular dynamics results showed that the active components of PFB, such as ellagic acid, luteolin, berberrubine, procyanidin B1, and adenosine, had better affinity with these key target genes. By qPCR and Western blot assays, we verified that the expressions of CDK1 and EZH2 and the key factors of the MPAK signaling pathway were significantly down-regulated in HCC.

CONCLUSION

This study demonstrated that the n-butanol extract of PFB exhibits a strong inhibitory effect on the proliferation of HepG2 cells and clarifies the underlying molecular mechanisms involved. By precisely modulating the expression levels of critical signaling molecules - including CDK1, PDGFRB, AKT1, FGFR1, MAPK1, and EZH2 - the n-butanol extract of PFB robustly disrupts cancer cell cycle progression and perturbs the activity of associated signaling pathways, thereby significantly curtailing the aberrant proliferation of tumor cells. This study not only elucidated the effects of the n-butanol extract of PFB on the aforementioned targets but also established a theoretical and experimental basis for further investigating their application in the treatment of HCC. Furthermore, it offers novel insights and research directions for the development of innovative therapeutic strategies derived from natural products, particularly those centered on multi-target synergistic approaches for liver cancer treatment.