COMPREHENSIVE LC-MS-DRIVEN IDENTIFICATION AND QUANTITATIVE CHARACTERIZATION OF BIOACTIVE ACETOGENINS FROM ANNONA MURICATA AND THEIR MODULATION OF ONCOGENIC SIGNALING IN TRIPLE-NEGATIVE BREAST CANCER: IN-VITRO ANTI-PROLIFERATIVE AND PRO-APOPTOTIC ASSESSMENT

Abstract

Annona muricata is a tropical plant producing bioactive annonaceous acetogenins with potent cytotoxic properties. In this study, acetogenin-rich fractions from A. muricata leaves were characterized using UHPLC–QToF-MS, revealing multiple structurally diverse major and minor constituents. Their effects were evaluated in normal and transformed mammalian cell lines, assessing cell viability, cell cycle progression, apoptosis, mitochondrial membrane potential, and oxidative stress. The acetogenin fraction caused a dose- and time-dependent reduction in cell viability, with transformed cells showing higher sensitivity. Flow cytometry demonstrated G₂/M phase–specific cell cycle arrest, accompanied by significant apoptosis, while fluorescence assays confirmed mitochondrial depolarization and increased reactive oxygen species, implicating intrinsic apoptotic pathways. Comparative analysis across mammalian models revealed conserved mitochondrial responses, alongside tissue-specific differences in susceptibility. These findings indicate that A. muricata acetogenins exert mitochondria-mediated cytotoxicity with conserved apoptotic mechanisms, emphasizing their relevance in comparative zoology and evolutionary toxicology, and highlighting the utility of integrating LC-MS phytochemical profiling with mammalian cell models for natural product–based biomedical research.