P38-β/SAPK-inhibiting and apoptosis-inducing activities of (E)-4-chloro-2-((3-ethoxy-2-hydroxybenzylidene) amino)phenol

Abstract

The present study has three purposes; first evaluating cytotoxicity of (E)-4-chloro-2-((3-ethoxy-2-hydroxybenzylidene)amino)phenol (ACES), second deciphering ACES-mediated cellular death mechanism, and third estimating ACES-mediated alterations in the expressions of mitogen-activated protein kinase (MAPK) pathway-related genes. Neutral red uptake assay, cell cycle analysis, mitochondrial membrane potential (MMP), reactive oxygen species (ROS) measurements, caspase 3/7 and 9 activations, and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) were implemented. IC50 values of ACES-treated five cells were around 4–6 µg/mL. However, Caco-2 and Huh-7 cells were found to be twofold resistant and fivefold sensitive with IC50 values of 11 µg/mL and 0.93 µg/mL, respectively. In this study, it was initially reported that ACES exhibits selective cytotoxicity to Huh-7 cells. In addition, ACES induced apoptosis by nuclear fragmentation, MMP disruption, and intracellular ROS elevation in MCF-7 cells. qRT-PCR experiment indicated the expressions of 30 genes including ATF2, CREB1, MYC, NFATC4 (NFAT3), CCNA1, CCNB1, CCND2, CDK2, CDKN1A (p21CIP1), CDKN1C (p57KIP2), CDKN2A (p16INK4a), CDKN2B (p15INK4b), DLK1, NRAS, CDC42, PAK1, MAP4K1 (HPK1), MAP3K3 (MEKK3), MAP2K3 (MEK3), MAP2K6 (MEK6), MOS, MAPK1 (ERK2), MAPK8 (JNK1), MAPK10 (JNK3), MAPK11 (p38-β), LAMTOR3 (MP1), MAPK8IP2 (JIP-1), PRDX6 (AOP2), COL1A1, and HSPA5 (Grp78) were downregulated at least 1.5-fold. Moreover, ACES effectively inhibited expressions of genes that code for elements of p38-β/stress-activated protein kinase (SAPK) pathway. ACES has the potential to be used for the reversal of trastuzumab resistance in breast cancer patients by inhibiting p38/SAPK pathway in MCF-7 cells. Therefore, with the selective cytotoxic, apoptosis-inducing, and p38-β/SAPK-inhibiting activities, ACES can be utilized for developing a novel anticancer drug.

The Gut Microbiota Metabolite Urolithin B Improves Cognitive Deficits by Inhibiting Cyt C-Mediated Apoptosis and Promoting the Survival of Neurons Through the PI3K Pathway in Aging Mice

“…JNK, also known as stress-activated protein kinase (SAPK) and p38, are activated by environmental stresses such as OS, and signaling pathways participate in mediating apoptosis in host cells upon injury or infection ( Kaneto et al, 2004 ). Furthermore, the presence and activation of JNK/p38 effectively promotes Cyt-C release from mitochondria into the cytosol, and JNK/p38-mediated Cyt-C release contributes to an increase in the activity of caspase-3 concomitantly with the onset of apoptosis ( Karaosmanolu, 2020 ). Furthermore, the activation of poly (ADP-ribose) polymerase 1 (PARP-1) in response to upregulation of p-JNK levels is another symbol of neuronal apoptosis ( Johnson and Nakamura, 2007 ).…”

The Gut Microbiota Metabolite Urolithin B Improves Cognitive Deficits by Inhibiting Cyt C-mediated Apoptosis and Promoting the Survival of Neurons Through the PI3K Pathway in Aging Mice

“…JNK, also known as stress-activated protein kinase (SAPK) and p38, are activated by environmental stresses such as OS, and signaling pathways participate in mediating apoptosis in host cells upon injury or infection [37]. Furthermore, the presence and activation of JNK/p38 effectively promotes Cyt-C release from mitochondria into the cytosol, and JNK/p38-mediated Cyt-C release contributes to an increase in the activity of caspase-3 concomitantly with the onset of apoptosis [38]. Furthermore, the activation of poly(ADP-ribose) polymerase 1 (PARP-1) in response to upregulation of p-JNK levels is another symbol of neuronal apoptosis [39].…”

Recent knowledge of NFATc4 in oncogenesis and cancer prognosis

“…To further explain EVs function, de Camargo et al demonstrated EVs from the NFATc4-expressing cells decreased the cell proliferation rate and induced cell apoptosis through macrophages recruitment in the 2D cell culture system [ 59 ]. In addition, NFATc4 down-regulated during (E)-4-chloro-2-((3-ethoxy-2-hydroxybenzylidene) amino) phenol (ACES) induced apoptosis through p38-β/SAPK in MCF-7 breast cancer cells, indicating that NFATc4 could be a latent cancer therapeutic target [ 60 ].…”