Calycosin Induces Gastric Cancer Cell Apoptosis via the ROS-Mediated MAPK/STAT3/NF-κB Pathway

Yu Zhang; Jian-Qiang Zhang; Tong Zhang; Hui Xue; Wen-Bo Zuo; Yan-Nan Li; Yue Zhao; Geng Sun; Zhong-Ren Fu; Qing Zhang; Xue Zhao; Yue Teng; An-Qi Wang; Jia-Zhu Li; Ying Wang; Cheng-Hao Jin

doi:10.2147/OTT.S292388

Calycosin Induces Gastric Cancer Cell Apoptosis via the ROS-Mediated MAPK/STAT3/NF-κB Pathway

Onco Targets Ther. 2021 Apr 12:14:2505-2517. doi: 10.2147/OTT.S292388. eCollection 2021.

Authors

Yu Zhang^#¹, Jian-Qiang Zhang^#^{2

3}, Tong Zhang¹, Hui Xue¹, Wen-Bo Zuo¹, Yan-Nan Li¹, Yue Zhao¹, Geng Sun¹, Zhong-Ren Fu¹, Qing Zhang¹, Xue Zhao¹, Yue Teng¹, An-Qi Wang¹, Jia-Zhu Li¹, Ying Wang^{4

5}, Cheng-Hao Jin^{1

4

5}

Affiliations

¹ Department of Biochemistry and Molecular Biology, College of Life Science & Technology, Heilongjiang Bayi Agricultural University, Daqing, People's Republic of China.
² Department of Food Science and Technology, College of Food Science, Northeast Agricultural University, Harbin, People's Republic of China.
³ Heilongjiang Heyi Dairy Technology Co. Ltd., Daqing, People's Republic of China.
⁴ Department of Food Science and Engineering, College of Food Science & Technology, Heilongjiang Bayi Agricultural University, Daqing, People's Republic of China.
⁵ National Coarse Cereals Engineering Research Center, Daqing, People's Republic of China.

^# Contributed equally.

Abstract

Background: Calycosin, an active compound in plants, can promote the apoptosis of various cancer cells; however, the mechanism by which it regulates reactive oxygen species (ROS) in gastric cancer (GC) cells remains unclear.

Purpose: In this study, we investigated the effects of calycosin on apoptosis, the cell cycle, and migration in GC cells under ROS regulation.

Results: The results of the Cell Counting Kit-8 assay suggested that calycosin had significant cytotoxic effects on 12 gastric cancer cells, but no significant cytotoxic effects on normal cells. Hoechst 33342/propidium iodide (PI) double staining and flow cytometry showed that calycosin had clear pro-apoptotic effects on AGS cells. Western blotting revealed that the expression of cytochrome C and pro-apoptotic proteins B-cell lymphoma 2 (Bcl-2)-associated agonist of cell death (Bad), cleaved (cle)-caspase-3, and cle-poly (ADP-ribose) polymerase gradually increased, and the expression of anti-apoptotic protein Bcl-2 gradually decreased. Calycosin also decreased the expression of extracellular signal-regulated kinase, nuclear factor kappa B (NF-κB), and signal transducer and activator of transcription 3 (STAT3), and increased the phosphorylation levels of p38, c-Jun N-terminal kinase, and inhibitor of NF-κB. In addition, calycosin markedly increased ROS accumulation, and pretreatment with active oxygen scavenger n-acetyl-l-cysteine (NAC) clearly inhibited apoptosis. Calycosin downregulated the cell cycle proteins cyclin-dependent kinase 2 (CDK2), CDK4, CDK6, cyclin D1, and cyclin E; upregulated p21 and p27; and arrested cells in the G0/G1 phase. Similarly, calycosin also downregulated Snail family transcriptional repressor 1, E-cadherin, and β-catenin and inhibited cell migration. However, pretreatment with NAC inhibited the calycosin-induced effects of cycle arrest and migration.

Conclusion: In summary, calycosin induces apoptosis via ROS-mediated MAPK/STAT3/NF-κB pathways, thereby exerting its anti-carcinogenic functions in GC cells.

Keywords: apoptosis; calycosin; cell cycle; cell migration; human gastric cancer; reactive oxygen species.

Grants and funding

This work was supported by the central government supports local college reform and development fund talent training projects, Heilongjiang Touyan Innovation Team Program (2019HTY078), the Project for Heilongjiang Bayi Agricultural University (XDB202012), the Heilongjiang Farms & Land Reclamation Administration Support Project for Key Scientific Research (HKKYZD190705), the Heilongjiang Bayi Agricultural University Support Program for “San Zong” (TDJH201905), Heilongjiang Province College Student Innovation and Entrepreneurship Training Program Project (202010223001), and Heilongjiang Province College Student Innovation and Entrepreneurship Training Program Project (202010223004).