Document Type : Pharmacology & Toxicology
Authors
1
Department of Pathobiology, SR.c., Islamic Azad University, Tehran, Iran
2
Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran;
3
3 Biotechnology Department, Pasteur Institute of Iran, Tehran, Iran.
4
Department of Basic Science, Faculty of Veterinary Medicine, Science and Research Branch, Islamic Azad University, Tehran, Iran;
Abstract
Background: Breast cancer remains a leading cause of cancer-related death in women, and resistance to standard treatments highlights the need for new therapeutic strategies. FINO2, a potent inducer of ferroptosis, and pirfenidone, an antifibrotic agent, have shown promising potential in targeting tumor survival and growth pathways. Objective: This study aimed to explore the antitumor effects of FINO2 and pirfenidone, both alone and combined, in a mouse model of breast cancer, focusing on ferroptosis induction, oxidative stress markers, and tumor microenvironment changes. Methods: The model was established using the 4T1 cell line in Balb/C mice. Cells were gently mixed with Phosphate-buffered saline (PBS), and 100 μl of this suspension (containing 1×10^5 4T1 cells) was injected either subcutaneously or into the mammary fat pad of the right flank of the mice, using a G28 insulin syringe. After injection, a 14-day delay was allowed to ensure tumor development. Tumor formation was confirmed after 7 to 10 days, when the tumor reached 0.5 cm ×0.5 cm. Mice with mammary tumors were then divided into treatment groups: FINO2, pirfenidone, doxorubicin, the combination of FINO2 and pirfenidone, and a control disease group. Tumor volume was measured, and qRT-PCR was used to assess Iron Regulatory Protein 2 (IRP2) gene expression related to ferroptosis. Histopathological analysis with trichrome staining was conducted to examine fibrosis, and oxidative stress levels were measured using the total oxidative stress (TOS) assay. Results: FINO2 significantly induced ferroptosis, demonstrated by changes in IRP2 and related genes, while pirfenidone reduced tumor-associated fibrosis. The combined treatment of FINO2 and pirfenidone produced the most notable effects, including the most significant reduction in tumor volume, increased oxidative stress, and decreased fibrosis compared to individual treatments and controls. Conclusion: The combination of FINO2 and pirfenidone effectively triggers ferroptosis and alters the tumor microenvironment, leading to reduced tumor growth and the potential to overcome drug resistance in breast cancer. These findings emphasize the clinical potential of this combined approach.
Keywords
Main Subjects
)