Investigating the Effects of Morin and Aminoimidazole Carboxamide Ribonucleotide on the AMPK/mTOR/ULK1 Signaling...

Breast cancer, one of the most prevalent cancers, remains the leading cause of mortality worldwide. Dysregulation of autophagy and apoptosis regulators is closely associated with cancer progression. AMP-activated protein kinase (AMPK) plays a critical role in tumorigenesis by regulating these processes. This study aimed to investigate the anticancer effects of Morin and aminoimidazole carboxamide ribonucleotide (AICAR) on viability, apoptosis, and autophagy in MCF-7 cells.

MCF-7 cells were treated with different concentrations of Morin and AICAR for 24, 48, and 72 hours. Cell viability was assessed using the MTT assay to determine the effective Morin dose. Protein levels of UNC-51-like kinase 1 (ULK1), AMPK, mammalian target of rapamycin (mTOR), and LC3B/LC3A were analyzed by Western blotting. Reactive oxygen species (ROS) production was measured, and mitochondrial membrane potential was evaluated using rhodamine 123 fluorescence.

The (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) MTT assay revealed that Morin (*P < 0.01), AICAR (*P < 0.01), and their combination (##P < 0.001) significantly reduced MCF-7 cell viability. Morin treatment markedly increased p-AMPK and p-ULK1 expression (*P < 0.05-***P < 0.001, #P < 0.05-##P < 0.01), and upregulated LC3B/LC3A (*P < 0.01-**P < 0.001, #P < 0.05), while p-mTOR expression was significantly downregulated (*P < 0.05-**P < 0.01, #P < 0.05). ROS generation in mitochondria increased markedly (*P < 0.01-**P < 0.001, #P < 0.01). Furthermore, Morin significantly decreased mitochondrial membrane potential (**P < 0.001-***P < 0.0001, #P < 0.01).

Morin, reduced MCF-7 cell survival and proliferation, enhanced apoptosis, increased ROS production, and diminished mitochondrial membrane potential, confirming its potential anticancer role.