Psoralidin inhibits stemness-associated characteristics in cervical cancer cells via modulation of Oct4/Sox2 expression.

Resistance to chemo/radio therapy, along with recurrence of aggressive tumors, is a major factor contributing to cervical cancer (CaCx) mortality. This is largely driven by a small subset of slow-growing cancer stem cells (CSC). The present study aimed to explore the potential of psoralidin to target CaCx-CSC. Molecular docking (MD) and MD simulation was conducted to analyze the interaction of psoralidin with core stemness markers Oct4, Sox2, and Nanog and stability of interaction, respectively. A low-adhesive culture system was established to enrich CSC, which were then treated with the IC50 concentration of psoralidin to study anti-CSC effect. Psoralidin demonstrated strong binding to stemness markers, cell permeability, and no predicted hepatotoxicity in silico. Binding stability was highest with Oct4, followed by Nanog and Sox2. IC50 values of 3-4 µM were observed across CaCx cell lines C33a, SiHa, and HeLa regardless of HPV status. Cervical-CSC enriched under low-adhesion conditions displayed elevated stemness marker expression, confirmed by qRT-PCR and immunoblotting. Psoralidin-treated cervicospheres showed reduced spheroid size, number, and structural integrity. Treatment also downregulated stemness markers as well as NF-κB p65, c-Rel, and pSTAT3(Y727). Overall, our preclinical findings demonstrate the potential of psoralidin to target CCSC and provide a strong rationale for its further translational evaluation as an adjuvant to existing therapeutic regimens in cervical cancer.