Enhanced efficacy of breast cancer treatment with etoposide-graphene oxide nanogels: A novel nanomedicine approach
Abbas Asoudeh-Fard
1,2 
, Milad Mohkam
3, Asghar Parsaei
4 , Shadi Asghari
5 , Antonio Lauto
6,7, Fatemeh Khoshnoudi
8 , Mustafa Mhmood Salman Al-Mamoori
9 , Mohadeseh Asoudeh-Fard
10, Hossine Ghasemi Sadabadi
11,12, Ahmad Gholami
13,14* 1 Institute Galilée-University Sorbonne, University Sorbonne Paris North, Paris, France
2 Research Center for Pharmaceutical Nanotechnology, Tabriz University of Medical Sciences, Tabriz, Iran
3 Allergy Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
4 Niko Gene Saba Company, Rayan Novin Pajoohan Pars, Biotechnology Company, Biotechnology Incubator, Shiraz University of Medicine Sciences, Shiraz, Iran
5 Department of Microbiology, Shiraz Branch Islamic Azad University, Shiraz, Iran
6 School of Science, University of Western Sydney, Campbelltown, NSW, 2560, Australia
7 School of Medicine, University of Western Sydney, Campbelltown, NSW, 2560, Australia
8 Department of Cellular and Molecular, Zarghan Branch Islamic Azad University, Zarghan, Iran
9 Department of Cellular and Molecular, Mashhad Branch Islamic Azad University, Mashhad, Iran
10 Department of General Medicine, Azad Zahedan Medicine University, Zahedan, Iran
11 Hematology and Oncology Research Center, Tabriz University of Medical Sciences Tabriz, Iran
12 Faculty of Medicine Tabriz, University of Medical Sciences Tabriz, Iran
13 Biotechnology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
14 Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran
Abstract
Introduction: Breast cancer represents a significant global health challenge, underscoring the need for innovative therapeutic strategies. This study explores the therapeutic potential of etoposide (ETO)-loaded graphene oxide (GO) nanogels to enhance the efficacy of breast cancer treatments.
Methods: ETO-GO nanogels were synthesized and characterized using field-emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), energy-dispersive X-ray spectroscopy (EDS), and Fourier-transform infrared spectroscopy (FT-IR). Cytotoxicity was evaluated through MTT assays on MCF-7 breast cancer cells and normal HUVEC cells. Apoptosis induction was assessed using DAPI staining, flow cytometry, and quantitative reverse transcription polymerase chain reaction (qRT-PCR) to analyze changes in gene expression.
Results: Characterization confirmed the formation of uniform, spherical nanogels with high ETO encapsulation efficiency. EDS and FT-IR analyses validated the successful loading of the drug onto the GO matrix. Cytotoxicity assays revealed a dose-dependent response, with significantly stronger effects observed in MCF-7 cells (20% viability at 100 µg/mL) than HUVEC cells (40% viability at the same concentration), indicating selective cytotoxicity. Apoptosis was verified through DAPI staining, which showed characteristics of nuclear fragmentation, and flow cytometry, identifying 15.35% of the treated cells as apoptotic. qRT-PCR analysis demonstrated an upregulation of pro-apoptotic genes (CASP3, CASP8, CASP9, BAX, PTEN) by as much as 8.3-fold, alongside a marked downregulation of the anti-apoptotic gene Bcl-2, confirming the potent induction of apoptosis by the nanogels.
Conclusion: ETO-GO nanogels show promising potential for targeted breast cancer therapy, providing enhanced drug delivery and selective cytotoxicity. These findings warrant further in vivo studies to validate their clinical applicability.