Morusin Attenuates Cancer Stemness and Modulates CSC-driven Macrophage Polarization in Cervical Cancer

INTRODUCTION: Cancer stem cells (CSC) role in treatment resistance is well established. CSCs promote M2 macrophage polarization within the tumor microenvironment, creating a pro-tumorigenic feedback loop that sustains CSC homeostasis. Therefore, disrupting this cross-talk can be an effective therapeutic strategy. In the present study, we identified Morusin as a potent inhibitor that could impact both cancer stemness and CSCinduced macrophage polarization. METHODS: In silico screening was performed to assess the druggability of Morusin and its binding to stemness markers. Molecular Docking and simulation were performed to check the stability of Morusin binding with markers. To evaluate its effect on cancer stemness, low-adhesion spheroids were treated with Morusin at IC50, followed by qPCR and western blot for stemness markers. To investigate Morusin's influence on CSC-mediated macrophage polarization, innovative transwell co-culture systems were employed to assess the impact of tumor spheroids on macrophage phenotype. RESULT: Molecular docking identified Morusin as a binder of the stemness factors Oct4, Sox2, and Nanog, with MD simulations confirming stable interactions and the strongest binding to Oct4. In an in vitro system, Morusin reduced spheroid size, disrupted spheroidal integrity and self-renewal, and lowered Oct4 and Sox2 levels, proving to be an effective anti-CSC agent. In a transwell system, activated THP1 cells co-cultured with 3D tumorspheres showed enhanced M2 polarization compared to monolayers, whereas Morusin-treated tumorspheres shifted THP1 cells toward an M1 phenotype. DISCUSSION: These findings indicate that Morusin targets both Cervical-CSC stemness and prevents CSCinduced M2 polarization, thus dampening CSC-Macrophage crosstalk. CONCLUSION: These findings indicate that Morusin targets both Cervical-CSC stemness and prevents CSCinduced M2 polarization, thus dampening CSC-Macrophage crosstalk.