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©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
18β-glycyrrhetinic acid inhibits proliferation of gastric cancer cells through regulating the miR-345-5p/TGM2 signaling pathway
Xia Li, Xiao-Ling Ma, Yi Nan, Yu-Hua Du, Yi Yang, Dou-Dou Lu, Jun-Fei Zhang, Yan Chen, Lei Zhang, Yang Niu, Ling Yuan
Xia Li, Yu-Hua Du, Ling Yuan, College of Pharmacy, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
Xiao-Ling Ma, Yi Nan, Yan Chen, Yang Niu, Traditional Chinese Medicine College, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
Yi Nan, Lei Zhang, Yang Niu, Key Laboratory of Hui Ethnic Medicine Modernization of Ministry of Education, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
Yi Yang, College of Basic Medicine, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
Dou-Dou Lu, Jun-Fei Zhang, College of Clinical Medicine, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
Author contributions: Li X carried out most of the studies, and analyzed the data; Li X, Ma XL, and Du YH wrote the manuscript; Ma XL and Du YH carried out the chart-making work; Yang Y, Lu DD, and Zhang JF were responsible for network pharmacology analysis; Chen Y and Niu Y designed the study and revised the manuscript; Yuan L provided the conceptual and technical guidance as well as revised the manuscript critically for important intellectual content; and all authors have read and approved the manuscript.
Supported by the Ningxia Natural Science Foundation, No. 2022AAC03144.
Institutional review board statement: The study was reviewed and approved by the Institutional Review Board of Ningxia Medical University (No.2021-N0063).
Institutional animal care and use committee statement: The animal protocol was approved by the Institutional Animal Care and Use Committee of Ningxia Medical University (IACUC-NYLAC-2022-108).
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: All data generated or analyzed during this study are included in this paper, and further inquiries can be directed to the corresponding author (20080017@nxmu.edu.cn).
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
https://creativecommons.org/Licenses/by-nc/4.0/ Corresponding author: Ling Yuan, MD, PhD, Assistant Professor, College of Pharmacy, Ningxia Medical University, No. 1160 Shengli Street, Yinchuan 750004, Ningxia Hui Autonomous Region, China. 20080017@nxmu.edu.cn
Received: March 20, 2023
Peer-review started: March 20, 2023
First decision: April 10, 2023
Revised: April 24, 2023
Accepted: May 17, 2023
Article in press: May 17, 2023
Published online: June 21, 2023
BACKGROUND
Gastric cancer (GC) is a common gastrointestinal malignancy worldwide. Based on cancer-related mortality, the current prevention and treatment strategies for GC still show poor clinical results. Therefore, it is important to find effective drug treatment targets.
AIM
To explore the molecular mechanism of 18β-glycyrrhetinic acid (18β-GRA) regulating the miR-345-5p/TGM2 signaling pathway to inhibit the proliferation of GC cells.
METHODS
CCK-8 assay was used to determine the effect of 18β-GRA on the survival rate of GES-1 cells and AGS and HGC-27 cells. Cell cycle and apoptosis were detected by flow cytometry, cell migration was detected by a wound healing assay, the effect of 18β-GRA on subcutaneous tumor growth in BALB/c nude mice was investigated, and the cell autophagy level was determined by MDC staining. TMT proteomic analysis was used to detect the differentially expressed autophagy-related proteins in GC cells after 18β-GRA intervention, and then the protein-protein interaction was predicted using STRING (https://string-db.org/). MicroRNAs (miRNAs) transcriptome analysis was used to detect the miRNA differential expression profile, and use miRBase (https://www.mirbase/) and TargetScan (https://www.targetscan.org/) to predict the miRNA and complementary binding sites. Quantitative real-time polymerase chain reaction was used to detect the expression level of miRNA in 18β-GRA treated cells, and western blot was used to detect the expression of autophagy related proteins. Finally, the effect of miR-345-5p on GC cells was verified by mir-345-5p overexpression.
RESULTS
18β-GRA could inhibit GC cells viability, promote cell apoptosis, block cell cycle, reduce cell wound healing ability, and inhibit the GC cells growth in vivo. MDC staining results showed that 18β-GRA could promote autophagy in GC cells. By TMT proteomic analysis and miRNAs transcriptome analysis, it was concluded that 18β-GRA could down-regulate TGM2 expression and up-regulate miR-345-5p expression in GC cells. Subsequently, we verified that TGM2 is the target of miR-345-5p, and that overexpression of miR-345-5p significantly inhibited the protein expression level of TGM2. Western blot showed that the expression of autophagy-related proteins of TGM2 and p62 was significantly reduced, and LC3II, ULK1 and AMPK expression was significantly increased in GC cells treated with 18β-GRA. Overexpression of miR-345-5p not only inhibited the expression of TGM2, but also inhibited the proliferation of GC cells by promoting cell apoptosis and arresting cell cycle.
CONCLUSION
18β-GRA inhibits the proliferation of GC cells and promotes autophagy by regulating the miR-345-5p/TGM2 signaling pathway.
Core Tip: Gastric cancer (GC) is a global health problem that seriously endangers human life, so it is urgent to find drugs to treat it. 18β-glycyrrhetinic acid (18β-GRA), as one of the main components of glycyrrhiza, has a strong antitumor effect. In this paper, the inhibitory effect of 18β-GRA on GC was verified by in vitro and in vivo experiments. In addition, it was found that 18β-GRA promoted autophagy and inhibited the proliferation of GC cells through the miR-345-5p/TGM2 signaling pathway. These findings provide the theoretical basis for the GC clinical treatment of 18β-GRA.
