Identification of novel molecular targets in triple negative breast cancer by meta analysis approach and validating the synergistic effect of anethole and doxorubicin in modulating the gene and protein expression in MDA MB 231 cells

Abstract

As triple negative breast cancer (TNBC) is very aggressive and lacks a newlinedruggable target, the present study aims to find an effective molecular target along with newlinea safe pharmaceutical lead to reduce the side effects of chemotherapy. In this study, an newlineintegrated bioinformatics and in vitro cell line studies were carried out to discover the key newlinecandidate genes specific for TNBC. The study was carried out into four distinct phases. newlineIn the phase I of the study, we have employed a meta-analysis approach to identify newlinegenes that are specific for TNBC. This was done by investigating the transcriptomic newlineprofiles of TNBC and compared the tumor samples with non-tumor samples from gene newlineexpression omnibus (GEO) to identify and validate consistently differentially expressed newlinegenes (CDGs) which are commonly observed among various TNBC microarray profiles newlineand highly specific to TNBC and the drug that could possibly modulate the expression of newlinethese genes. In phase II of the study, we have assessed the ability of the phytocompound newlineanethole and the standard drug, which was obtained from the results of phase I namely, newlinedoxorubicin for its anticancer activity in MDA-MB-231 cell line by assessing its newlineantiproliferative effect, inhibition of colony formation and migration and cellular newlinelocalization using Mito Tracker and Lyso Tracker. In phase III of the study, the mode of newlinecell death executed by the synergistic action of anethole and doxorubicin was evaluated newlineby various parameters namely apoptosis, cell cycle analysis, changes in mitochondrial newlinemembrane potential, expression levels of apoptotic and anti-apoptotic proteins, ROS, ER newlinestress and multiparametric analysis to confirm apoptosis, DNA damage, inhibition of cell newlineproliferation was done using various antibodies. In phase IV of the study, we attempted newlineto evaluate the ADME properties of anethole and interaction of doxorubicin and anethole newlinewith the CDGs and associated pathway genes which were obtained from the phase I newlineresults by in silico docking studies using Schrodinger drug designing suite