The in vitro assessment of the prooxidative action of the novel aroylacrylic acid phenylamides in A549 and MCF7 cancer cell lines

Abstract

Aroylacrylic acids, their phenylamides and other analogues bearing structural scaffold comprising of the two phenyl rings linked by three or four carbon atoms exert prominent anticancer activity. To unravel the relying mechanism responsible for induction of cell death by aroylacrylic acid phenylamides, we designed four novel structural analogues ( 1a - 1d ) by means of Michaels’ addition of secondary cyclic amines to aroylacrylic acid phenylamides. A battery of biological assays has been utilized to evaluate functional and morphological alterations triggered by the investigated treatments in A549 and MCF7 cell lines that are KRAS^mut and KRAS^wt, respectively, using carbonyl cyanide- p -trifluoromethoxyphenylhydrazone as a positive control. Concomitant treatment with, N -acetylcysteine reversed cell death response induced by investigated compounds (75.7 ± 15.1 % and 33.1 ± 9.9 % in average for MCF7 and A549 cells, respectively), suggesting that the generation of reactive oxidative species (ROS) was the key trigger of the cell death. Moreover, MCF7 cells were more susceptible to experimental treatments than A549 cells, whereas examination of the mitochondrial membrane potential and superoxide generation showed that the integrity of mitochondria in MCF7 cell line was devastated quickly after exposure to the investigated compounds. As KRAS^mut A549 cells have dysfunctional oxidative phosphorylation (OXPHOS), these results imply that the fully operative OXPHOS in KRAS^wt cells exceedingly amplified ROS production upon treatment. Additionally, ultrastructural analysis confirmed significant mitochondrial damage in both cell lines. Investigated compounds potently disrupt mitochondrial function and can serve as a valuable asset for testing capacity of new antioxidant agents.