Investigation of the mechanism of euphornin against cervical cancer using network pharmacology.

Euphornin has potential therapeutic effect on cervical cancer (CC). To elucidate the mechanism of euphornin in treating CC, the strategy of combining network pharmacology and in vitro cell experiments was adopted. Network pharmacology analysis was conducted to obtain the core targets and key pathways associated with euphornin's therapeutic action against CC. Molecular docking tools were employed to analyze the affinity between euphornin and its core targets. Western blot was applied to verify the core target proteins and the key signaling pathways. Wound healing and transwell assays were performed to assess the migration and invasion of CC cells, and flow cytometry was conducted to evaluate CC cells' rate of apoptosis. Ten core targets of euphornin in treating CC were identified, and enrichment analysis revealed their involvement in apoptosis, miRNA metabolism/transcription/regulation, and p53 pathways. It was demonstrated by molecular docking analysis that there was a high affinity between euphornin and ESR1. Compared to the control group, euphornin significantly inhibited CC cell migration and invasion, accompanied by decreased expression of EGFR, MDM2, MMP-2, and MMP-9 (P < 0.05), along with increased expression of caspase3, ESR1, p-p53 and p53 (P < 0.05). Compared to the pcDNA3.1 group, the transfected pcDNA3.1 with ESR1 exhibited a remarkable increase in ESR1 expression (P < 0.0001), and significant inhibition on CC cell migration and invasion (P < 0.001). In addition, the apoptosis rate of CC cells and the expression of cleaved-caspase3, cleaved-caspase8, and cleaved-caspase9 increased significantly (P < 0.001). Compared to the euphornin + si-NC group, the migration and invasion of Hela cells in the euphornin + si-ESR1 group increased significantly (P < 0.01), while their apoptosis decreased significantly (P < 0.001). Our study identifies targets of euphornin in treating CC and proposes that its mechanism potentially involves the regulation of the MDM2-p53 signaling pathway. ESR1 was revealed to be a potential anti-CC target of euphornin, and euphornin was demonstrated to inhibit CC cell migration and invasion, and promote CC cell apoptosis by targeting ESR1.