Abstract
BACKGROUND: Universally, Allium ascalonicum (Shallots) is a well-known flavouring agent in many cuisines. Though, it's been proved for its health benefits due to the presence of alkaloids, flavonoids, terpenoids, phenols and coumarins, its role as an anti-neoplastic agent still requires comprehensive investigation. In our study, we have investigated the presence of potential anti-neoplastic phytocompounds, anti-inflammatory, cytotoxicity and anti-metastatic activity of Shallots against Triple-Negative Breast Cancer cell line, MDA-MB-231. METHODS: Phytocompounds of aqueous Allium ascalonicum extract (AAE) derived from GC-MS and LC-MS analysis were docked with an inflammatory marker, Interleukin-18 (IL-18); anti-apoptotic proteins, B-cell Lymphoma-2 (BCL-2) and Myeloid Cell Leukemia-1 (MCL-1); and metastatic marker, Vimentin using PyRx (Version 0.9.9). Subsequently, the anti-inflammatory property of AAE was determined using Bovine Serum Albumin (BSA) Denaturation Assay and the chemotherapeutic potential of AAE was determined using 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay on MDA-MB-231 and HEK293T cell lines. Additionally, to determine the synergistic effect of Doxorubicin Hydrochloride (Standard) and AAE, MTT assay was performed on MDA-MB-231 cell lines treated with the combination therapy. Furthermore, the anti-metastatic property of AAE was determined using cell migration and clonogenic assays. Finally, Dual Acridine Orange/Ethidium Bromide fluorescence staining method was used to determine if AAE has the ability to induce apoptosis and necrosis in MDA-MB-231 cells. RESULTS: Molecular docking results using the compounds obtained from LC-MS and GC-MS with the target proteins revealed promising anti-neoplastic bioactive compounds. BSA Denaturation assay proved that AAE has anti-inflammatory property, with the highest, 85.78% observed at 2 mg/ml of AAE. Moreover, MTT assay proved that AAE exhibited cytotoxic effect on MDA-MB-231 in a dose-dependent manner, with an IC(50) observed at 1.23 mg/ml (**p ≤ 0.005) and non-toxic to HEK293T cells. Combination therapy of the standard with AAE reduced the IC(50) of the standard by 65.5%. Consecutively, the anti-metastatic property of AAE was proved using cell migration and clonogenic assays, suggesting suppression of epithelial-mesenchymal transition. Finally, Dual Acridine Orange/Ethidium Bromide fluorescence staining method displayed that, AAE has the ability to induce apoptosis and necrosis in TNBC cells. CONCLUSION: The outcomes from in vitro assays corroborated with the molecular docking results and hence, on authenticating the potentiality of AAE's anti-neoplastic effect via. in vivo models, pre-clinical and clinical trials, Allium ascalonicum can be articulated to a prospective anti-neoplastic drug for treating TNBC.