Investigating the Inhibitory Aspects of Metformin/Curcumin Co-Treatment through Convergence of In-Silico and In-Vitro Approaches

Abstract

Background: Nearly 16% of people with breast cancer (BC) have Diabetes Mellitus type 2 (DM2) and are at a higher risk of death worldwide. However, their common regulatory factors and functional mechanisms have not been profoundly understood. In this study, we used in silico approaches to study their co-regulated differentially expressed genes (DEGs) in order to explore and introduce novel therapeutic targets.\nResults: Microarray analysis identified 196 co-regulated DEGs (156 upregulated and 40 downregulated) between BC and DM2. The most significant pathway was predicted to be Fc gamma R-mediated phagocytosis through pathway analysis. Through protein-protein interaction network investigation based on the co-regulated DEGs and performing topological characterization, common hubs were revealed. Then hubs9 regulatory factors including TFs and miRNAs were also mined to introduce unprecedented therapeutic targets. According to Gene Ontology analysis, cell death and migration related processes found to be important enough for designing in vitro experiments. Co-treatment of Metformin (MTFN) and Curcumin (CURC) was the main inhibitory strategy we adopted as they both have impact on BC and DM2. Drug treatment was performed on two BC cell lines (MCF7 and MDA-MB-231). Combination index calculation by MTT viability assay proved additive effects and slight synergism on both cell lines. The superior apoptotic potential of co-treatment compared to single treatments was shown on inhibition of MCF7 proliferation and induction of cell death demonstrated by cell body co-staining and flow cytometry as well as gene expression analysis via RT-PCR. Furthermore, wound-healing scratch assay showed that this co-treatment has a slightly higher effect on migration inhibition compared to single treatments. \nConclusion: In conclusion, our study used in silico approaches and introduced a regulatory panel between BC and DM2. We also proved that our co-treatment inhibitory strategy successfully controlled the biological processes under investigation.