Zeaxanthin Induces Apoptosis via ROS-Regulated MAPK and AKT Signaling Pathway in Gastric Cancer Cells

Background: Zeaxanthin, a carotenoid commonly found in plants, has a variety of biological functions including anti-cancer activity. Purpose: This study aimed to investigate the potential mechanisms of zeaxanthin in human gastric cancer cells. Methods: CCK-8 assay was used to examine the cytotoxic effect of zeaxanthin on human gastric cancer cells. Flow cytometry was used to analyse AGS cell cycle distribution and apoptosis status. Western blot analysis was used to detect the expression levels of cyclerelated proteins (Cyclin A, Cyclin B1, CDK1/2, p21, and p27), apoptosis-related proteins (Bcl-2, Bad, caspase-3, PARP), MAPK, AKT, STAT3, and NF-kappa B. Results: CCK-8 assay showed that zeaxanthin has obvious cytotoxic effects on 12 types of human gastric cancer cells, but no obvious toxic effect on normal cells. In addition, flow cytometry and Western blotting results showed that zeaxanthin induces apoptosis by reducing mitochondrial membrane potential; increasing Cytochrome C, Bax, cleaved-caspase-3 (decas-3), and cleaved-PARP (cle-PARP) expression levels; and decreasing Bcl-2, pro-caspase-3 (pro-cas-3), and pro-PARP expression levels. Additionally, zeaxanthin caused cell cycle arrest at the G2/M phase by increasing the levels of p21 and p27 and reduced the levels of AKT, Cyclin A, Cyclin Bl, and Cyclin-dependent kinase 1/2 (CDK1/2). Furthermore, after zeaxanthin treatment, the expression levels of reactive oxygen species (ROS), p-JNK, p-p38, and I-kappa B increased, and the expression levels of p-ERK, p-AKT, STAT3, and NF-kappa B decreased. However, the ROS scavenger N-acetylcysteine (NAC) and MAPK inhibitors inhibited zeaxanthin-induced apoptosis, and under the action of zeaxanthin, MAPK regulated NF-kappa B and STAT3, and reduced their protein expression levels. Conclusion: Zeaxanthin has a potential effect against gastric cancer cells through the ROSmediated MAPK, AKT, NF-kappa B, and STAT3 signaling pathways, and it is expected to become a new drug for the treatment of human gastric cancer.