玉米黄质通过活性氧调节的丝裂原活化蛋白激酶和蛋白激酶B信号通路诱导人胃癌细胞凋亡。

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

作者信息

Sheng Ya-Nan, Luo Ying-Hua, Liu Shao-Bin, Xu Wan-Ting, Zhang Yu, Zhang Tong, Xue Hui, Zuo Wen-Bo, Li Yan-Nan, Wang Chang-Yuan, Jin Cheng-Hao

机构信息

Department of Food Science and Engineering, College of Food Science, Heilongjiang Bayi Agricultural University, Daqing 163319, People's Republic of China.

Department of Grass Science, College of Animal Science & Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing 163319, People's Republic of China.

出版信息

Onco Targets Ther. 2020 Oct 29;13:10995-11006. doi: 10.2147/OTT.S272514. eCollection 2020.

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 cycle-related proteins (Cyclin A, Cyclin B1, CDK1/2, p21, and p27), apoptosis-related proteins (Bcl-2, Bad, caspase-3, PARP), MAPK, AKT, STAT3, and NF-κB.

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 (cle-cas-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 B1, 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-κB increased, and the expression levels of p-ERK, p-AKT, STAT3, and NF-κB decreased. However, the ROS scavenger N-acetylcysteine (NAC) and MAPK inhibitors inhibited zeaxanthin-induced apoptosis, and under the action of zeaxanthin, MAPK regulated NF-κB and STAT3, and reduced their protein expression levels.

CONCLUSION

Zeaxanthin has a potential effect against gastric cancer cells through the ROS-mediated MAPK, AKT, NF-κB, and STAT3 signaling pathways, and it is expected to become a new drug for the treatment of human gastric cancer.

背景

玉米黄质是一种常见于植物中的类胡萝卜素，具有多种生物学功能，包括抗癌活性。

目的

本研究旨在探讨玉米黄质对人胃癌细胞作用的潜在机制。

方法

采用CCK-8法检测玉米黄质对人胃癌细胞的细胞毒性作用。运用流式细胞术分析AGS细胞周期分布及凋亡状态。采用蛋白质免疫印迹法检测细胞周期相关蛋白（细胞周期蛋白A、细胞周期蛋白B1、细胞周期蛋白依赖性激酶1/2、p21和p27）、凋亡相关蛋白（Bcl-2、Bad、半胱天冬酶-3、聚（ADP-核糖）聚合酶）、丝裂原活化蛋白激酶（MAPK）、蛋白激酶B（AKT）、信号转导子和转录激活子3（STAT3）以及核因子κB（NF-κB）的表达水平。

结果

CCK-8法检测显示，玉米黄质对12种人胃癌细胞具有明显的细胞毒性作用，但对正常细胞无明显毒性作用。此外，流式细胞术和蛋白质免疫印迹结果表明，玉米黄质通过降低线粒体膜电位诱导细胞凋亡；增加细胞色素C、Bax、切割的半胱天冬酶-3（cle-cas-3）和切割的聚（ADP-核糖）聚合酶（cle-PARP）的表达水平；降低Bcl-2、原半胱天冬酶-3（pro-cas-3）和原聚（ADP-核糖）聚合酶的表达水平。此外，玉米黄质通过增加p21和p27的水平使细胞周期阻滞于G2/M期，并降低AKT、细胞周期蛋白A、细胞周期蛋白B1和细胞周期蛋白依赖性激酶1/2（CDK1/2）的水平。此外，玉米黄质处理后，活性氧（ROS）、磷酸化的应激活化蛋白激酶（p-JNK）、磷酸化的p38丝裂原活化蛋白激酶（p-p38）和核因子κB抑制蛋白（I-κB）的表达水平升高，而磷酸化的细胞外信号调节激酶（p-ERK）、磷酸化的AKT、STAT3和NF-κB的表达水平降低。然而，ROS清除剂N-乙酰半胱氨酸（NAC）和MAPK抑制剂抑制了玉米黄质诱导的细胞凋亡，并且在玉米黄质作用下，MAPK调节NF-κB和STAT3，并降低它们蛋白的表达水平。