Effects of Compound in Hedyotis diffusa Willd against Acute Myeloid Leukemia: An In silico and In vitro Study

Background: Hedyotis diffusa Willd (HDW) is an herb that has been used empirically for treating cancer, and its antileukemic effect has been confirmed by laboratory evidence. This study aimed to explore the underlying mechanism by which HDW and its active compound exert effects on acute myeloid leukemia (AML) through in silico analyses combined with experimental validation. Methods: The targets of the compounds were collected from the database and intersected with AML targets. Based on these data, a protein-protein interaction (PPI) network and compound-target (C-T) network were constructed, and KEGG enrichment analysis was performed. Topological analysis of the C-T network and PPI network was performed to screen for hub compounds and targets. Molecular dynamics simulations were conducted to test the binding mode and strength between the targets and the compounds at the molecular level. Cell viability, flow cytometry, ELISA, and Q-PCR were further used to evaluate the in silico results. Results: A total of 86 targets of 12 screened active compounds of HDW against AML were identified. According to topological analysis, tumor protein p53 (TP53) and signal transducer and activator of transcription 3 (STAT3) exhibited the highest degree of centrality (DC) in the PPI networks of HDW targets. Quercetin had a higher affinity for TP53 than for STAT3. Molecular dynamics simulations confirmed that the TP53-quercetin docked complex was stable with respect to the original TP53-ligand complex. The targets of HDW and quercetin against AML were significantly enriched in multiple biological processes, including the p53 signaling pathway and apoptosis. The results from the in vitro experiment confirmed that quercetin triggers apoptosis in the human AML cell line KG-1 through the p53 pathway protein. Conclusion: This study outlines the multi-compound, multi-target, and multi-pathway mechanism by which HDW affects AML based on an in silico predictive model and further validates the antileukemic mechanism of the screened active compound in an in vitro model. This study provides a perspective for studying the antileukemic mechanism of HDW for further research.