Basic Study
Copyright ©The Author(s) 2021. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Mar 14, 2021; 27(10): 939-958
Published online Mar 14, 2021. doi: 10.3748/wjg.v27.i10.939
Huanglian decoction suppresses the growth of hepatocellular carcinoma cells by reducing CCNB1 expression
Min Li, Hua Shang, Tao Wang, Shui-Qing Yang, Lei Li
Min Li, Hua Shang, Department of Gastroenterology, Zibo Central Hospital, Zibo 255036, Shandong Province, China
Tao Wang, Department of General Surgery, Zibo Central Hospital, Zibo 255036, Shandong Province, China
Shui-Qing Yang, Lei Li, School of Biological Science and Medical Engineering, Beihang University, Beijing 100191, China
Lei Li, Department of Pathology, University of Otago, Dunedin px806, New Zealand
Author contributions: Li L conceived and performed the experiments and drafted the article; Li M and Shuang H conceived and performed partial experiments; Wang T and Yang SQ designed the study, supervised the experiments, and revised the article; all authors read and approved the final manuscript.
Institutional review board statement: Animal and human experiments were not conducted in this study.
Conflict-of-interest statement: The authors declare that there are no competing interests associated with the manuscript.
Data sharing statement: Technical appendix, statistical code, and dataset are available from the corresponding author at [wshililei@buaa.edu.cn].
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: http://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Lei Li, PhD, Academic Research, Department of Pathology, University of Otago, 270 Great King Street, Dunedin px806, New Zealand. wshililei@buaa.edu.cn
Received: November 26, 2020
Peer-review started: November 26, 2020
First decision: December 21, 2020
Revised: January 3, 2021
Accepted: February 1, 2021
Article in press: February 1, 2021
Published online: March 14, 2021
Processing time: 105 Days and 3.1 Hours
Abstract
BACKGROUND

Hepatocellular carcinoma (HCC) is one of the most prevalent cancers in human populations worldwide. Huanglian decoction is one of the most important Chinese medicine formulas, with the potential to treat cancer.

AIM

To investigate the role and mechanism of Huanglian decoction on HCC cells.

METHODS

To identify differentially expressed genes (DEGs), we downloaded gene expression profile data from The Cancer Genome Atlas Liver Hepatocellular Carcinoma and Gene Expression Omnibus (GSE45436) databases. We obtained phytochemicals of the four herbs of Huanglian decoction from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform. We also established a regulatory network of DEGs and drug target genes and subsequently analyzed key genes using bioinformatics approaches. Furthermore, we conducted in vitro experiments to explore the effect of Huanglian decoction and to verify the predictions. In particular, the CCNB1 gene was knocked down to verify the primary target of this decoction. Through the identification of the expression levels of key proteins, we determined the primary mechanism of Huanglian decoction in HCC.

RESULTS

Based on the results of the network pharmacological analysis, we revealed 5 bioactive compounds in Huanglian decoction that act on HCC. In addition, a protein-protein interaction network analysis of the target genes of these five compounds as well as expression and prognosis analyses were performed in tumors. CCNB1 was confirmed to be the primary gene that may be highly expressed in tumors and was significantly associated with a worse prognosis. We also noted that CCNB1 may serve as an independent prognostic indicator in HCC. Moreover, in vitro experiments demonstrated that Huanglian decoction significantly inhibited the growth, migration, and invasiveness of HCC cells and induced cell apoptosis and G2/M phase arrest. Further analysis showed that the decoction may inhibit the growth of HCC cells by downregulating the CCNB1 expression level. After Huanglian decoction treatment, the expression levels of Bax, caspase 3, caspase 9, p21 and p53 in HCC cells were increased, while the expression of CDK1 and CCNB1 was significantly decreased. The p53 signaling pathway was also found to play an important role in this process.

CONCLUSION

Huanglian decoction has a significant inhibitory effect on HCC cells. CCNB1 is a potential therapeutic target in HCC. Further analysis showed that Huanglian decoction can inhibit HCC cell growth by downregulating the expression of CCNB1 to activate the p53 signaling pathway.

Keywords: Huanglian decoction (Coptidis Rhizoma, Zingiberis Rhizoma, Folium Artemisiae Argyi, Mume Fructus); Hepatocellular carcinoma; The Cancer Genome Atlas; Gene Expression Omnibus; P53 pathway; Cell cycle; Apoptosis

Core Tip: The purpose of this study was to investigate the role and mechanism of Huanglian decoction on hepatocellular carcinoma (HCC) cells. The results showed that Huanglian decoction has a significant inhibitory effect on the growth of HCC cells. Huanglian decoction can inhibit HCC cell growth by downregulating the expression of CCNB1 to activate the p53 signaling pathway. CCNB1 is a potential therapeutic target for liver cancer.