Modulatory Effects of Metformin Alone and in Combination with Cimetidine and Ibuprofen on T Cell-related Parameters in a Breast Cancer Model

Fereshteh Taghipour; Omolbanin Oladpour; Mohammad Taghi Rezayati; Hossein Khorramdelazad; Maryam Nemati; Zahra Taghipour; Javad Masoumi; Zuhair Mohammad Hassan; Abdollah Jafarzadeh; Abdollah Jafarzadeh

doi:10.18502/ijaai.v20i5.7410

Fereshteh Taghipour Department of Immunology, School of Medicine, Rafsanjan University of Medical Sciences, Rafsanjan, Iran
Omolbanin Oladpour Department of Immunology, School of Medicine, Rafsanjan University of Medical Sciences, Rafsanjan, Iran
Mohammad Taghi Rezayati Molecular Medicine Research Center, Research Institute of Basic Medical Sciences, Rafsanjan University of Medical Sciences, Rafsanjan, Iran
Hossein Khorramdelazad Molecular Medicine Research Center, Research Institute of Basic Medical Sciences, Rafsanjan University of Medical Sciences, Rafsanjan, Iran
Maryam Nemati Department of Hematology and Laboratory Sciences, School of Para-Medicine, Kerman University of Medical Sciences, Kerman, Iran
Zahra Taghipour Department of Histology, School of Medicine, Rafsanjan University of Medical Sciences, Rafsanjan, Iran
Javad Masoumi Molecular Medicine Research Center, Research Institute of Basic Medical Sciences, Rafsanjan University of Medical Sciences, Rafsanjan, Iran
Zuhair Mohammad Hassan Department of Immunology, School of Medicine, Tarbiat Modarres University, Tehran, Iran
Abdollah Jafarzadeh Molecular Medicine Research Center, Research Institute of Basic Medical Sciences, Rafsanjan University of Medical Sciences, Rafsanjan, Iran
Abdollah Jafarzadeh Molecular Medicine Research Center, Research Institute of Basic Medical Sciences, Rafsanjan University of Medical Sciences, Rafsanjan, Iran

DOI: https://doi.org/10.18502/ijaai.v20i5.7410

Keywords: Breast neoplasms; Cimetidine; Ibuprofen; Metformin; Mice; T-lymphocytes

Abstract

Metformin, cimetidine, and ibuprofen separately exhibit immunomodulatory and anti-tumorigenic effects. Herein, the impacts of metformin alone and in combination with cimetidine/ibuprofen on some Th1- and regulatory T (Treg) cell-related parameters were evaluated using a breast cancer (BC) model.

For establishing the BC model, four groups of Balb/c mice were challenged with the carcinoma cell line. After 11-30 days post-induction, they were treated intraperitoneally (with metformin (200 mg/kg), "metformin plus cimetidine (20 mg/kg)"; "metformin plus ibuprofen (20 mg/kg)", or with all three drugs in mentioned doses. Untreated BC and without tumor mice were enrolled as control groups. On day 31, splenic Th1 and Treg cell frequencies, serum interferon-gamma (IFN-γ), and transforming growth factor-beta (TGF-β) concentration, and intra-tumoral T-bet, TGF-β, and forkhead box protein P3 (FOXP3) expression were measured; using flow cytometry, enzyme-linked immunosorbent assay (ELISA), and real-time-PCR, respectively.

Treatment of the BC mice with metformin alone and in combination with cimetidine and/or ibuprofen enhanced the frequency of Th1 cells, and IFN-γ concentration, while it resulted in a decrease in the frequency of Treg cells, serum TGF-β concentration, and the expression of FOXP3 and TGF-β compared with un-treated BC mice. FOXP3 expression in the metformin-treated group was lower in mice who received combination therapy. Survival rate and body weight were increased, while tumor size and spleen index were reduced in mice treated with metformin alone and its combination with cimetidine and/or ibuprofen. No remarkable differences were found between metformin-treated mice and those who received combination therapies regarding Th1 and Treg cell percentages, TGF-β expression, body weight, tumor size, and spleen index.

The benefits of combinational therapy may be largely attributed to metformin. Immunotherapeutic potentials of metformin in cancers need further considerations.