Background/Aim: Breast cancer remains one of the most commonly diagnosed cancers among women worldwide and is a significant public health concern. Aim of this study was to investigate the in vitro anticancer activity of Opuntia cochenillifera ethanolic extract (EOC) against triple-negative breast cancer (TNBC) cells and to predict potential molecular targets involved in its mechanism of action through bioinformatics analysis.Methods: Fresh O cochenillifera leaves were extracted by ethanol maceration. Phytochemical constituents were analysed using qualitative screening, thin-layer chromatography (TLC), UV-Vis spectrophotometry and Fourier transform infrared (FTIR) spectroscopy. Cytotoxicity against MDA-MB-231 TNBC cells was determined via MTT assay and morphological assessment. Apoptosis induction was quantified by Annexin V-FITC/PI flow cytometry. Potential molecular targets were identified through integrated bioinformatics platforms, including STITCH, SEA, SwissTargetPrediction, STRING and Cytoscape, followed by pathway enrichment analysis.Results: Phytochemical analysis revealed the presence of alkaloids, flavonoids, phenolics, saponins and steroids, with flavonoids as predominant constituents (68 mg/g extract). FTIR spectra confirmed hydroxyl, aliphatic, aromatic and carbonyl functional groups consistent with phenolic and flavonoid structures. EOC exhibited dose-dependent cytotoxicity against MDA-MB-231 cells, with an IC50 of 270.63 µg/mL, accompanied by morphological and flow-cytometric evidence of apoptosis. Network pharmacology analysis identified 34 apoptosis-related target genes, including six hub genes (CDKN1A/p21, TP53, MAPK1, MAPK3, AKT1 and FOXO3), associated with TP53-p21 signalling, MAPK cascades and mitochondrial stress-induced apoptosis pathways.Conclusion: O cochenillifera ethanolic extract demonstrated significant cytotoxic and pro-apoptotic effects in TNBC cells, potentially mediated through TP53-p21 and MAPK pathway modulation. These findings suggest that O cochenillifera is a promising candidate for further investigation as a natural adjuvant therapeutic agent for triple-negative breast cancer.
Alaydrus et al. (Thu,) studied this question.