G-3664

Determining the synergistic effect of magnetic field on apoptosis rate and expression of Bax/Bcl2 by folic acid targeted iron nanoparticles loaded Thymoquinon on Breast cancer cell line Mcf7

 Mohammad Zamani Rarani ¹, Reza Afzalipour ¹ ©, Mohammad Erfan Shafiee ², Mehdi Bahjati Ardakani ³, Amin Ghanbarnejad ⁴, Sayed Reza Mirlohi ⁵ ℗, Niloofar Choobin ⁵   

 
Abstract

Introduction: Breast cancer is one of the most prevalent and fatal cancers among women, and its effective management remains a global challenge. In recent years, nanomedicine has emerged as a promising avenue for improving cancer therapy through targeted drug delivery. Among nanocarriers, superparamagnetic iron oxide nanoparticles (SPIONs) are of particular interest due to their biocompatibility, magnetic responsiveness, and ability to be functionalized for targeted therapy. Apoptosis is a key mechanism in cancer treatment, regulated by genes such as Bax (pro-apoptotic) and Bcl2 (anti-apoptotic). This study investigates the synergistic apoptotic effects of thymoquinone (TQ), a natural anticancer compound, when delivered via folic acid-targeted SPIONs, with and without magnetic field exposure, in MCF-7 breast cancer cells. Methods and Materials: Superparamagnetic iron oxide nanoparticles were synthesized using the thermal decomposition method and characterized by TEM and FTIR at the University of Isfahan. The following experimental groups were established: (1) untreated control, (2) TQ-only, (3) nanoparticle-only, (4) drug-loaded nanoparticles, (5) folic acid-targeted nanoparticles, and (6) folic acid-targeted drug-loaded nanoparticles. MCF-7 cells were cultured in DMEM/F12 medium and treated at various time points (24, 48, 72 hours). TQ and CPA (adenosine A1 receptor agonist) were obtained from Sigma. Cell viability was measured via MTT assay, and apoptosis was analyzed using Annexin V/PI flow cytometry. Bax and Bcl2 gene expression was quantified by real-time PCR using the ΔΔCt method. Results: MCF-7 cells exhibited typical morphology and adhered well during culture. TQ at 25 μM significantly reduced cell viability in a time-dependent manner, with the lowest viability observed at 72 hours. Flow cytometry revealed a marked increase in early and late apoptotic cells following TQ treatment, while no significant apoptosis was observed in DMSO controls. The IC50 values were determined to be 87 nmol for TQ and 180 μmol for CPA at 24 hours. Real-time PCR showed that Bax expression increased with longer TQ exposure, peaking at 72 hours, while Bcl2 expression decreased significantly. Targeted nanoparticles loaded with TQ showed enhanced apoptotic effects compared to non-targeted groups, especially under magnetic field application (P 0.05). Conclusion and Discussion: The combination of folic acid-targeted SPIONs and thymoquinone demonstrated a strong pro-apoptotic effect in MCF-7 cells, mediated by upregulation of Bax and downregulation of Bcl2. These findings highlight the potential of magnetic nanoparticle-based targeted delivery systems as a powerful tool in breast cancer therapy, especially when combined with natural compounds and external magnetic fields to enhance treatment efficacy.

Keywords: Magnetic field, apoptosis, thymoquinone, folic acid, iron, MCF7