Exploring the ROS-mediated anti-cancer potential in human triple-negative breast cancer by garlic bulb extract: A source of therapeutically active compounds
Shivbrat Upadhyay, Rumana Ahmad, Raushan Kumar, Sneha Ghildiyal, Alok Singh, Khursheed Ahmad, Ishrat Husain, Md. Abul Barkat, Mohd. Zaheen Hassan, Yahya I. Asiri, Sahabjada Siddiqui
Abstract
Allium sativum L. has been used medicinally and traditionally since antiquity. This study sought to examine the Allium sativum ethanolic extract (ASEE) in inducing apoptosis in human triple-negative breast cancer (TNBC) MDA-MB-231 cells and the molecular interactions of the identified components with cell death markers using in silico method. Cytotoxicity was tested on MCF-7, MDA-MB-231, and Normal Vero cells. Further, ROS production, apoptosis study, MMP, cell cycle by flow cytometer, and western blot analysis were also performed. ASEE was phytochemically characterized by the HPLC while AutoDock Vina and iGEMDOCK performed in-silico binding interaction studies. The HPLC method identified two active organosulfur chemicals, allicin and alliin, in ASEE. MTT test revealed significant (p < 0.05) inhibition of breast cancer cells proliferation. The inhibitory effect of ASEE was more pronounced in MDA-MB-231 cells than in MCF-7 cells, however, no substantial cytotoxicity was seen in normal Vero cells. TNBC cells treated with high concentrations of ASEE were found in the late apoptotic stage and exhibited an increase in ROS level and a reduction in MMP. ASEE exposure increased the percentage of cells in the G2/M phase. ASEE upregulated the p53 and Bax proteins while downregulated the Bcl-2, p-Akt, and p-p38 proteins. Allicin and alliin compounds had strong binding affinity with targeted proteins of breast cancer, as demonstrated by molecular docking. Both compounds had good pharmacokinetics and druglikeness. ASEE could be useful in the treatment of human triple-negative breast cancer without any safety risks.