Copyright
©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
Thioridazine reverses trastuzumab resistance in gastric cancer by inhibiting S-phase kinase associated protein 2-mediated aerobic glycolysis
Zheng-Yan Yang, Yi-Wei Zhao, Jing-Rui Xue, Ran Guo, Zhi Zhao, Han-Di Liu, Zhi-Guang Ren, Ming Shi
Zheng-Yan Yang, Ran Guo, Han-Di Liu, Zhi-Guang Ren, Department of Pathology, School of Basic Medical Sciences, Henan University, Kaifeng 475004, Henan Province, China
Yi-Wei Zhao, Jing-Rui Xue, Joint National Laboratory for Antibody Drug Engineering, Henan University, Kaifeng 475004, Henan Province, China
Zhi Zhao, Department of Pathology, Henan University-affiliated Zhengzhou Yihe Hospital, Zhengzhou 450000, Henan Province, China
Zhi-Guang Ren, Key Laboratory of Clinical Resources Translation, The First Affiliated Hospital, Henan University, Kaifeng 475004, Henan Province, China
Ming Shi, Center of Clinical Oncology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou 221002, Jiangsu Province, China
Ming Shi, Jiangsu Center for the Collaboration and Innovation of Cancer Biotherapy, Xuzhou Medical University, Xuzhou 221004, Jiangsu Province, China
Co-first authors: Zheng-Yan Yang and Yi-Wei Zhao.
Co-corresponding authors: Zhi-Guang Ren and Ming Shi.
Author contributions: Yang ZY drafted the manuscript and conducted the experiments of immunoblotting, qPCR, and apoptosis analysis in this study; Zhao YW completed the glucose metabolism analysis and animal experiments; Xue JR contributed to the CCK-8 assay and animal experiments; Guo R assisted in the construction and amplification of recombinant plasmid vectors; Liu HD participated in the data collection of docking; Zhao Z assisted in immunohistochemical analysis of xenograft tumor tissue samples; Ren ZG supervised the experiments, corrected the data, and revised the manuscript; Shi M conceived and designed the main content of this study and was responsible for analyzing the research results; all authors contributed to the article and approved the submitted version.
Supported by Youth Fund of National Natural Science Foundation of China, No. 81803575, and No. 31902287; Kaifeng Science and Technology Development Plan Project, No. 2203008; Key Specialized Research and Promotion Project of Henan Province in 2023, No. 232102311205; Henan Medical Science and Technology Research Program Project, No. LHGJ20210801; College Students Innovation and Entrepreneurship Training Program of Henan University, No. 20231022007.
Institutional review board statement: The study was reviewed and approved by the Ethics Committee of the Medical School of Henan University (HUSOM2022-452).
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 Medical School of Henan University (protocol number: HUSOM2022-439).
Conflict-of-interest statement: The authors declare no conflict of interest.
Data sharing statement: Relevant research data have been presented in the text. All data will be provided upon request if necessary.
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: Zhi-Guang Ren, Doctor, Associate Professor, Department of Pathology, School of Basic Medical Science, Henan University, No. 1 Jinming Street, Kaifeng 475004, Henan Province, China. renzhiguang66@outlook.com
Received: September 17, 2023
Peer-review started: September 17, 2023
First decision: October 8, 2023
Revised: October 19, 2023
Accepted: November 17, 2023
Article in press: November 17, 2023
Published online: December 7, 2023
Processing time: 74 Days and 14.9 Hours
ARTICLE HIGHLIGHTS
Research background
The drug resistance observed in patients with human epidermal growth factor receptor 2 (HER-2)-positive advanced gastric cancer (GC) treated with trastuzumab is a significant concern, as no established targeted therapy regimen for use after the development of drug resistance is available. S-phase kinase associated protein 2 (Skp2) has been identified as a crucial target for GC treatment; however, the development of new drugs targeting Skp2 remains a considerable challenge.
Research motivation
To investigate potential pharmacological interventions targeting Skp2 to increase the efficacy of subsequent therapies for patients with HER-2-positive GC who have developed resistance to trastuzumab.
Research objectives
This study aims to elucidate the inhibitory effect of thioridazine on Skp2 expression and to preliminarily assess the potential of thioridazine in reversing the resistance of HER2-positive GC cells to trastuzumab through both in vivo and in vitro experiments.
Research methods
The impact of altering the Skp2 protein expression level through overexpression or knockdown on the sensitivity of HER2-positive GC cells to trastuzumab was assessed using a cell counting kit-8 assay. The influence of thioridazine on Skp2 protein expression was demonstrated through computational docking analysis and Cellular Thermal Shift Assay. Flow cytometry, a glucose uptake assay, a lactate production assay, and xenograft experiments in nude mice were employed to evaluate the effects of thioridazine alone or in combination with trastuzumab and lapatinib on the cell cycle, apoptosis, glucose metabolism, and tumor growth.
Research results
Trastuzumab sensitivity can be increased in HER-2-positive GC cells through negative modulation of Skp2 expression. Thioridazine can selectively inhibit Skp2 expression and the protein kinase B/mammalian target of rapamycin signaling pathway. Thioridazine combined with lapatinib effectively reverses trastuzumab resistance in GC cells by diminishing glycolysis.
Research conclusions
Combining thioridazine with lapatinib is a potential strategy to reverse trastuzumab resistance in GC by suppressing Skp2 expression.
Research perspectives
Further investigation into the optimal combination ratio, initial dosage, and dose-response correlation between thioridazine and lapatinib in GC xenograft models will contribute to the development of more precise drug reference protocols for subsequent clinical trials. A new therapeutic strategy for the management of GC by simultaneous targeting of Skp2 and HER-2 could be introduced.
