Basic Study
Copyright ©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jul 21, 2023; 29(27): 4317-4333
Published online Jul 21, 2023. doi: 10.3748/wjg.v29.i27.4317
18β-glycyrrhetinic acid promotes gastric cancer cell autophagy and inhibits proliferation by regulating miR-328-3p/signal transducer and activator of transcription 3
Yi Yang, Yi Nan, Yu-Hua Du, Shi-Cong Huang, Dou-Dou Lu, Jun-Fei Zhang, Xia Li, Yan Chen, Lei Zhang, Ling Yuan
Yi Yang, Yu-Hua Du, Shi-Cong Huang, Dou-Dou Lu, Xia Li, Ling Yuan, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
Yi Nan, Jun-Fei Zhang, Yan Chen, Lei Zhang, Key Laboratory of Ningxia Minority Medicine Modernization Ministry of Education, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
Author contributions: Yang Y carried out most of the studies, analyzed the data and wrote the manuscript; Nan Y designed the study and revised the manuscript; Nan Y, Du YH and Huang SC wrote the manuscript and carried out the chart-making work; Lu DD and Li X were responsible for the total transcriptomic and bioinformatics analyses; Zhang JF performed parts of the in vivo and in vitro experiments and conducted statistical analyses of the data; Yuan L supervised the process of research and provided clinical guidance; Yuan L, Zhang L and Chen Y provided the conceptual and technical guidance as well as revised the manuscript critically for important intellectual content; All authors have read and approved the manuscript.
Supported by Ningxia Medical University Project, No. XZ2021005; Ningxia Natural Science Foundation, Nos. 2022AAC03144 and 2022AAC02039; and National Natural Science Foundation of China, No. 82260879.
Institutional review board statement: The study was reviewed and approved by the Institutional Review Board of Ningxia Medical University (No. 2021-X003, No. 2021-N0063, Nos. 2021-N001 and 2022-G089).
Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of the Ningxia Medical University (IACUC-NYLAC-2022-108).
Conflict-of-interest statement: All the authors report having 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, Professor, Ningxia Medical University, No. 1160 Shengli Street, Yinchuan 750004, Ningxia Hui Autonomous Region, China. nxykdx@qq.com
Received: March 28, 2023
Peer-review started: March 28, 2023
First decision: April 27, 2023
Revised: May 10, 2023
Accepted: June 2, 2023
Article in press: June 2, 2023
Published online: July 21, 2023
Processing time: 106 Days and 22.9 Hours
ARTICLE HIGHLIGHTS
Research background

Gastric cancer(GC) is among the most severe gastrointestinal malignancies, with high morbidity and mortality. In recent years, increasing evidence has shown that natural products can prevent and inhibit the development of GC by regulating miRNA and other genes, showing great therapeutic potential.

Research motivation

Targeting miRNA and other genes in GC with natural drugs is a promising strategy, which provides a valid reference to further elucidate the molecular mechanism of natural products in the treatment of GC.

Research objectives

The purpose of this study was to investigate the molecular mechanism of 18β-glycyrrhetinic acid (18β-GRA) regulating the miR-328-3p/ signal transducer and activator of transcription 3 (STAT3) signaling pathway, promoting the autophagy flow of GC cells and inhibiting cell proliferation.

Research methods

The differentially expressed miRNAs were screened by full transcriptomic analysis. The cells were transfected with lentivirus, and the functional indices of the cells were determined by Cell Counting Kit-8, clone forming method, and flow cytometry. The effect of overexpression of miR-328-3p on the tumorigenicity of GC cells was detected in the transplanted tumor model of nude mice. Hematoxylin-eosin staining and immunohistochemistry were used to observe tumor tissue morphology and detect protein expression, respectively. Bioinformatics analysis was performed using TransmiR, STRING, and miRWalk databases. Real-time quantitative polymerase chain reaction and western blot were used to detect mRNA and protein expression levels. Dual luciferase reporter system was used to verify the targeting relationship between genes. The autophagy flow of monomeric red fluorescent protein- green fluorescent protein - light chain 3 adenovirus double-labeled infected cells was observed under confocal laser microscopy.

Research results

18β-GRA could upregulate the expression of miR-328-3p in AGS cells. Overexpression of miR-328-3p inhibited the cell proliferation and colony formation ability, arrested the cell cycle, promoted apoptosis, inhibited the growth of subcutaneous tumors, and led to GC tissue cell necrosis increase. miR-328-3p can target and regulate STAT3. 18β-GRA intervention in GC cells and overexpression of miR-328-3p could downregulate the expression level of STAT3 mRNA. Compared with the vector group, the expression level of LC3 II was downregulated in the overexpressed miR-328-3p + bafilomycin A1 (Baf A1) group and upregulated in the overexpressed miR-328-3p + Baf A1 group. The number of yellow spots and separate red signals in the 18β-GRA group were significantly increased compared with the negative control group, and the number of yellow spots in the 18β-GRA + Baf A1 group was significantly increased compared with the Baf A1 group.

Research conclusions

18β-GRA promotes GC cells' autophagosome synthesis and inhibits cell proliferation by regulating the miR-328-3p/STAT3 signaling pathway.

Research perspectives

MiR-328-3p/STAT3 can be regulated by 18β-GRA and can be used as an effective drug target for the prevention and treatment of GC, thus providing a scientific basis for the clinical treatment of GC.