Modulation of β‐Catenin/ EMT Axis by Croton caudatus and Its Phytoflavonoid Kaempferol Ameliorates Cervical Cancer Migration and Stemness

Abstract

ABSTRACT Major gynecological malignancy like cervical cancer faces global challenges in conventional treatments, leading researchers to explore newer anticancer therapeutics with reduced toxicity and off‐target effects. Traditional ethnomedicinal plants with source of novel adjuvant chemotherapeutics are attracting considerable interest. β‐catenin pathway dysregulation is cardinal in cancer propagation, with well‐noted regulation in migration/EMT‐potential and cancer cell stemness. Present study focuses towards identifying molecular mechanisms of anti‐migratory efficacy of Croton caudatus leaf‐extract and its bioflavonoid Kaempferol against metastatic cervical cancer cells. C. caudatus leaf‐extract exhibited most cytotoxicity towards cervical cancer HeLa and SiHa cells. Leaf‐extract and Kaempferol significantly reduced cervical cancer cellular migration via downregulating EMT markers (N‐cadherin, Vimentin, Twist, Zeb2, Snail, Slug) and upregulating epithelial marker E‐cadherin ( p < 0.0001). In silico investigation depicted β‐catenin's importance in cervical cancer patient‐cohorts and its correlation with major EMT markers. C. caudatus leaf‐extract and Kaempferol altered EMT by inhibiting β‐catenin's nuclear translocation in vitro via upregulating 14‐3‐3 and downregulating Importin β‐1 ( p < 0.0001). Also they had upregulated upstream GSK3β and Phospho‐β‐catenin expression, demonstrating their anti‐migratory potential, most of which are imparted by extract's Kaempferol, at low IC 50 dose. Both leaf‐extract and Kaempferol significantly reduced expression of downstream c‐MYC, Cyclin D1, MMP‐7, and MMP‐9 by reducing β‐catenin's nuclear translocation ( p < 0.0001). Moreover, Kaempferol reduced expression of major stemness factors OCT4 and SOX2 in cervical cancer cells and decreased their physical association with β‐catenin. Thus, current study uncovers unknown pharmacological roles of C. caudatus leaf‐extract and Kaempferol in modulating β‐catenin's subcellular localization,altering EMT, thereby reducing cancer migration and stemness, encouraging their future use as alternative anti‐cancer therapeutics.