Evaluation of agmatine’s anti-cancer efficacy in Caco-2 colorectal adenocarcinoma cells

Abstract

This study aimed to evaluate the potential effects of agmatine on cell viability, migration, invasion, apoptosis, and the expression of the ABCB1, ABCC1, and ABCG2 genes in the Caco-2 colon cancer cell line. Agmatine efficacy was assessed thruogh proliferation, migration, and invasion assays at various concentrations. The apoptotic index was determined using apoptosis-related markers (Bax, Bcl-2, Csp-3) via apoptosis assays, quantitative real-time PCR (qRT-PCR), and Western blot analysis. Expression levels of the ABCG2, ABCB1, and ABCC1 genes were measured by qRT-PCR in agmatine-treated Caco-2 cells. Oxidative stress markers, including glutathione peroxidase (GPx) and catalase (CAT), were evaluated by qRT-PCR. Cell viability analysis revealed that agmatine exerted its most pronounced effects at 72 h, with significant reductions at concentrations of 6, 7.3, and 9 mM in Caco-2 cells and 6, 6.25, and 9 mM in L929 cells (p < 0.05). At these concentrations, migration and invasion assays showed dose-dependent decreases in cell motility and invasiveness in Caco-2 cells. Apoptosis analysis demonstrated a significant increase in the apoptotic index with rising agmatine concentrations. Significant decreases in GPx and CAT were observed in all three agmatine-treated Caco-2 groups compared to untreated controls (p < 0.01). However, the expression levels of ABCG2, ABCB1, and ABCC1 showed no significant changes following agmatine treatment (p > 0.05). These findings indicate that agmatine exerts antiproliferative, anti-migratory, anti-invasive, and pro-apoptotic effects in Caco-2 colon cancer cells, potentially through the modulation of apoptosis- and oxidative stress–related pathways. The lack of significant impacts on ABC transporter gene expression suggests that agmatine may be a promising candidate molecule for further translational studies in colorectal cancer.