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

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.