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©The Author(s) 2025. Published by Baishideng Publishing Group Inc. All rights reserved.
SEL1L-mediated endoplasmic reticulum associated degradation inhibition suppresses proliferation and migration in Huh7 hepatocellular carcinoma cells
Jia-Nan Chen, Li Wang, Yu-Xin He, Xiao-Wei Sun, Long-Jiao Cheng, Ya-Nan Li, Sei Yoshida, Zhong-Yang Shen
Jia-Nan Chen, Zhong-Yang Shen, School of Medicine, Nankai University, Tianjin 300071, China
Jia-Nan Chen, Zhong-Yang Shen, Department of Organ Transplant, Tianjin First Central Hospital, School of Medicine, Nankai University, Tianjin 300192, China
Li Wang, Yu-Xin He, Xiao-Wei Sun, Long-Jiao Cheng, Ya-Nan Li, Sei Yoshida, State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Frontiers Science Center for Cell Responses, Nankai University, Tianjin 300071, China
Sei Yoshida, Zhong-Yang Shen, Research Institute of Transplant Medicine, Nankai University, Tianjin 300192, China
Sei Yoshida, Nankai International Advanced Research Institute, Shenzhen 518045, Guangdong Province, China
Zhong-Yang Shen, Tianjin Key Laboratory for Organ Transplantation, Tianjin First Central Hospital, School of Medicine, Nankai University, Tianjin 300192, China
Co-corresponding authors: Sei Yoshida and Zhong-Yang Shen.
Author contributions: Chen JN, Wang L, He YX, Sun XW, Cheng LJ and Li YN performed the research; Chen JN and Wang L analyzed the data; Yoshida Sei and Shen ZY designed the study; All authors have read and approved the final manuscript.
Supported by the National Natural Science Foundation of China, No. 82241219, No. 82127808 and No. 81921004; and The Shenzhen Science and Technology Program, No. JCYJ20210324120813037.
Institutional review board statement: This study does not involve any human experiments.
Institutional animal care and use committee statement: Our research obtained ethical approval from the Ethics committee at Nankai University (approval No. 2025-SYDWLL-000002).
Conflict-of-interest statement: The authors declare that they have no conflict of interest.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Data sharing statement: No additional data are available.
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: Zhong-Yang Shen, MD, Professor, School of Medicine, Nankai University, No. 94 Fukang Road, Nankai District, Tianjin 300071, China.
zhongyangshen@nankai.edu.cn
Received: November 11, 2024
Revised: January 4, 2025
Accepted: February 5, 2025
Published online: March 14, 2025
Processing time: 107 Days and 22.3 Hours
BACKGROUND
Proteins play a central role in regulating biological functions, and various pathways regulate their synthesis and secretion. Endoplasmic reticulum-associated protein degradation (ERAD) is crucial for monitoring protein synthesis and processing unfolded or misfolded proteins in actively growing tumor cells. However, the role of the multiple ERAD complexes in liver cancer remains unclear.
AIM
To elucidate the effects of SEL1L-mediated ERAD on Huh7 and explore the underlying mechanisms in vivo and in vitro.
METHODS
Huh7 cells were treated with ERAD inhibitor to identify ERAD’s role. Cell counting kit-8, 5-ethynyl-2’-deoxyuridine and colony formation experiments were performed. Apoptosis level and migration ability were assessed using fluorescence activated cell sorting and Transwell assay, respectively. Huh7 SEL1L knockout cell line was established via clustered regularly interspaced short palindromic repeats, proliferation, apoptosis, and migration were assessed through previous experiments. The role of SEL1L in vivo and the downstream target of SEL1L were identified using Xenograft and mass spectrometry, respectively.
RESULTS
The ERAD inhibitor suppressed cell proliferation and migration and promoted apoptosis. SEL1L-HRD1 significantly influenced Huh7 cell growth. SEL1L knockout suppressed tumor cell proliferation and migration and enhanced apoptosis. Mass spectrometry revealed EXT2 is a primary substrate of ERAD. SEL1L knockout significantly increased the protein expression of EXT2. Furthermore, EXT2 knockdown partially restored the effect of SEL1L knockout.
CONCLUSION
ERAD inhibition suppressed the proliferation and migration of Huh7 and promoted its apoptosis. EXT2 plays an important role and ERAD might be a potential treatment for Huh7 hepatocellular carcinoma.
Core Tip: The endoplasmic reticulum-associated degradation (ERAD) inhibitor suppressed cell proliferation and migration and promoted apoptosis. SEL1L-HRD1 significantly influenced Huh7 cell growth. SEL1L knockout suppressed tumor cell proliferation and migration, and enhanced apoptosis. Mass spectrometry revealed EXT2 is a primary substrate of ERAD. SEL1L knockout significantly increased the protein expression of EXT2, whereas EXT2 knockdown partially restored the effect of SEL1L knockout. In conclusion, this study showed the function and possible mechanisms of ERAD in hepatocellular carcinoma, providing new insights into more effective treatment and strategies for liver cancer.
