Basic Study
Copyright ©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jul 21, 2022; 28(27): 3435-3454
Published online Jul 21, 2022. doi: 10.3748/wjg.v28.i27.3435
RING finger and WD repeat domain 3 regulates proliferation and metastasis through the Wnt/β-catenin signalling pathways in hepatocellular carcinoma
Ruo-Peng Liang, Xiao-Xue Zhang, Jie Zhao, Qin-Wei Lu, Rong-Tao Zhu, Wei-Jie Wang, Jian Li, Kai Bo, Chi-Xian Zhang, Yu-Ling Sun
Ruo-Peng Liang, Jie Zhao, Qin-Wei Lu, Rong-Tao Zhu, Wei-Jie Wang, Jian Li, Kai Bo, Chi-Xian Zhang, Yu-Ling Sun, Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China
Xiao-Xue Zhang, Department of Physical Examination, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China
Author contributions: Sun YL conceived the experiments; Liang RP, Zhang XX, Zhao J, Lu QW, Zhu RT, Wang WJ, Li J, Bo K and Zhang CX contributed sample collection/reagents/materials/analysis tools, performed all experiments, and analyzed the data; Liang RP, Zhang XX and Zhao J analyzed the results and wrote the paper; all authors have read and approve the final manuscript.
Supported by National Natural Science Foundation of China, No. 82172944 and No. 81900558; Co-operation Research Plan of Medical Science and Technology of Henan Province, No. LHGJ20190149; and The Key Scientific Research Projects of Universities of Henan Province, No. 21A320052.
Institutional review board statement: This study was reviewed and approved by the First Affiliated Hospital of Zhengzhou University, Zhengzhou, China, No. 10[2017].
Institutional animal care and use committee statement: All animal experimental protocols were conducted following the ARRIVE guidelines (Animal Research: Reporting In Vivo Experiments, and approved by Laboratory Animals and the Institutional Animal Care and Use Committee of the First Affiliated Hospital of Zhengzhou University (Approval Number: 10[2017]).
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: The data are available from the corresponding author on reasonable request.
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:
Corresponding author: Yu-Ling Sun, MD, PhD, Chief Doctor, Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Zhengzhou University, No. 50 West Jianshe Road, Zhengzhou 450052, Henan Province, China.
Received: February 7, 2022
Peer-review started: February 7, 2022
First decision: April 5, 2022
Revised: April 16, 2022
Accepted: June 3, 2022
Article in press: June 3, 2022
Published online: July 21, 2022

Hepatocellular carcinoma (HCC) exhibits high invasiveness and mortality rates, and the molecular mechanisms of HCC have gained increasing research interest. The abnormal DNA damage response has long been recognized as one of the important factors for tumor occurrence and development. Recent studies have shown the potential of the protein RING finger and WD repeat domain 3 (RFWD3) that positively regulates p53 stability in response to DNA damage as a therapeutic target in cancers.


To investigate the relationship between HCC and RFWD3 in vitro and in vivo and explored the underlying molecular signalling transduction pathways.


RFWD3 gene expression was analyzed in HCC tissues and adjacent normal tissues. Lentivirus was used to stably knockdown RFWD3 expression in HCC cell lines. After verifying the silencing efficiency, Celigo/cell cycle/apoptosis and MTT assays were used to evaluate cell proliferation and apoptosis. Subsequently, cell migration and invasion were assessed by wound healing and transwell assays. In addition, transduced cells were implanted subcutaneously and injected into the tail vein of nude mice to observe tumor growth and metastasis. Next, we used lentiviral-mediated rescue of RFWD3 shRNA to verify the phenotype. Finally, the microarray, ingenuity pathway analysis, and western blot analysis were used to analyze the regulatory network underlying HCC.


Compared with adjacent tissues, RFWD3 expression levels were significantly higher in clinical HCC tissues and correlated with tumor size and TNM stage (P < 0.05), which indicated a poor prognosis state. RFWD3 silencing in BEL-7404 and HCC-LM3 cells increased apoptosis, decreased growth, and inhibited the migration in shRNAi cells compared with those in shCtrl cells (P < 0.05). Furthermore, the in vitro results were supported by the findings of the in vivo experiments with the reduction of tumor cell invasion and migration. Moreover, the rescue of RFWD3 shRNAi resulted in the resumption of invasion and metastasis in HCC cell lines. Finally, gene expression profiling and subsequent experimental verification revealed that RFWD3 might influence the proliferation and metastasis of HCC via the Wnt/β-catenin signalling pathway.


We provide evidence for the expression and function of RFWD3 in HCC. RFWD3 affects the prognosis, proliferation, invasion, and metastasis of HCC by regulating the Wnt/β-catenin signalling pathway.

Keywords: RING finger and WD repeat domain 3, Hepatocellular carcinoma, Invasion, Proliferation, Metastasis, Wnt/β-catenin signaling pathways

Core Tip: The relationship between RING finger and WD repeat domain 3 (RFWD3) and tumorigenesis process has been reported occasionally, but the relationship between RFWD3 and hepatocellular carcinoma (HCC) is still unclear. This research reports the significant role of RFWD3 in HCC development by using bioinformatics databases, clinical samples, cell phenotypes, in vivo experiments, and microarray analyses. It provides evidence that RFWD3 expression might affect the tumorigenesis process of HCC by regulating the Wnt/β-catenin signalling pathways, laying a basic foundation for future relevant studies.