Basic Study
Copyright ©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastrointest Oncol. Oct 15, 2024; 16(10): 4244-4263
Published online Oct 15, 2024. doi: 10.4251/wjgo.v16.i10.4244
Clinical significance of upregulated Rho GTPase activating protein 12 causing resistance to tyrosine kinase inhibitors in hepatocellular carcinoma
Xiao-Wei Wang, Yu-Xing Tang, Fu-Xi Li, Jia-Le Wang, Gao-Peng Yao, Da-Tong Zeng, Yu-Lu Tang, Bang-Teng Chi, Qin-Yan Su, Lin-Qing Huang, Di-Yuan Qin, Gang Chen, Zhen-Bo Feng, Rong-Quan He
Xiao-Wei Wang, Yu-Xing Tang, Fu-Xi Li, Jia-Le Wang, Gao-Peng Yao, Da-Tong Zeng, Yu-Lu Tang, Qin-Yan Su, Lin-Qing Huang, Gang Chen, Zhen-Bo Feng, Department of Pathology, The First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
Da-Tong Zeng, Department of Pathology, Red Cross Hospital of Yulin City, Yulin 537000, Guangxi Zhuang Autonomous Region, China
Bang-Teng Chi, Rong-Quan He, Department of Medical Oncology, The First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
Di-Yuan Qin, Department of Computer Science and Technology, School of Computer and Electronic Information, Guangxi University, Nanning 530004, Guangxi Zhuang Autonomous Region, China
Co-first authors: Xiao-Wei Wang and Yu-Xing Tang.
Co-corresponding authors: Zhen-Bo Feng and Rong-Quan He.
Author contributions: Yao GP, Tang YL, Chen G, Feng ZB, and He RQ conceived and designed the study; Wang XW, Tang YX, Li FX, Wang JL, Zeng DT, Chi BT, Su QY, Huang LQ, and Qin DY performed the experiments, and analyzed the data; Wang XW, Tang YX, Li FX, Wang JL, Yao GP, Zeng DT, Chi BT, and Qin DY wrote the manuscript; Tang YL, Chen G, Feng ZB, and He RQ revised and corrected the draft; and all authors approved the final version of the article. Wang XW and Tang YX are co-first authors, contributing equally in study design, draft writing and data analysis. Wang XW also performed the immunohistochemical experiments and evaluated the corresponding results. Tang YX was responsible for computational pathology, including the calculation of standardized mean difference and summary receiver operating characteristic, as well as the analysis of related pathways. Feng ZB and He RQ are co-corresponding authors, contributing equally in designing the project, supervising various experiments and data mining, and correcting the draft.
Supported by National Natural Science Foundation of China, No. 82260581; Guangxi Zhuang Autonomous Region Health Committee Scientific Research Project, No. Z20201147; Guangxi Medical University Education and Teaching Reform Project, No. 2021XJGA02; Undergraduate Teaching Reform Project of Guangxi Higher Education, No. 2023JGB163; Guangxi Medical University Teacher Teaching Ability Development Project, No. 2202JFA20; and China Undergraduate Innovation and Entrepreneurship Training Program, No. S202310598170.
Institutional review board statement: The study was reviewed and approved by the Medical Ethics Committee of the First Affiliated Hospital of Guangxi Medical University Institutional Review Board, Approval No. 2022-KT-Guoji-127.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: Dataset available from the co-first author at tang_yx_patho@163.com. Participants gave informed consent for data sharing.
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: Zhen-Bo Feng, MD, PhD, Professor, Department of Pathology, The First Affiliated Hospital of Guangxi Medical University, No. 6 Shuangyong Road, Nanning 530021, Guangxi Zhuang Autonomous Region, China. fengzhenbo_gxmu@163.com
Received: June 24, 2024
Revised: August 17, 2024
Accepted: September 6, 2024
Published online: October 15, 2024
Processing time: 94 Days and 3.1 Hours
Abstract
BACKGROUND

Hepatocellular carcinoma (HCC) is a major health challenge with high incidence and poor survival rates in China. Systemic therapies, particularly tyrosine kinase inhibitors (TKIs), are the first-line treatment for advanced HCC, but resistance is common. The Rho GTPase family member Rho GTPase activating protein 12 (ARHGAP12), which regulates cell adhesion and invasion, is a potential therapeutic target for overcoming TKI resistance in HCC. However, no studies on the expression of ARHGAP12 in HCC and its role in resistance to TKIs have been reported.

AIM

To unveil the expression of ARHGAP12 in HCC, its role in TKI resistance and its potential associated pathways.

METHODS

This study used single-cell RNA sequencing (scRNA-seq) to evaluate ARHGAP12 mRNA levels and explored its mechanisms through enrichment analysis. CellChat was used to investigate focal adhesion (FA) pathway regulation. We integrated bulk RNA data (RNA-seq and microarray), immunohistochemistry and proteomics to analyze ARHGAP12 mRNA and protein levels, correlating with clinical outcomes. We assessed ARHGAP12 expression in TKI-resistant HCC, integrated conventional HCC to explore its mechanism, identified intersecting FA pathway genes with scRNA-seq data and evaluated its response to TKI and immunotherapy.

RESULTS

ARHGAP12 mRNA was found to be highly expressed in malignant hepatocytes and to regulate FA. In malignant hepatocytes in high-score FA groups, MDK-[integrin alpha 6 (ITGA6) + integrin β-1 (ITGB1)] showed specificity in ligand-receptor interactions. ARHGAP12 mRNA and protein were upregulated in bulk RNA, immunohistochemistry and proteomics, and higher expression was associated with a worse prognosis. ARHGAP12 was also found to be a TKI resistance gene that regulated the FA pathway. ITGB1 was identified as a crossover gene in the FA pathway in both scRNA-seq and bulk RNA. High expression of ARHGAP12 was associated with adverse reactions to sorafenib, cabozantinib and regorafenib, but not to immunotherapy.

CONCLUSION

ARHGAP12 expression is elevated in HCC and TKI-resistant HCC, and its regulatory role in FA may underlie the TKI-resistant phenotype.

Keywords: Hepatocellular carcinoma; Focal adhesion; Tyrosine kinase inhibitor; Rho GTPase activating protein 12; Drug resistance; Molecular mechanism; Biomarker

Core Tip: Molecular therapy has the potential to reverse tyrosine kinase inhibitors (TKIs) resistance in hepatocellular carcinoma (HCC). We combined 22861 single-cell RNA sequencing malignant cells, 5113 bulk RNA data (bulk RNA-seq and microarrays), and 562 immunohistochemistry samples to explore Rho GTPase activating protein 12 (ARHGAP12) high expression from mRNA and protein. The higher its expression, the worse the prognosis of HCC patients. HCC TKIs showed ARHGAP12 resistance. Mechanistically, we confirmed ARHGAP12 can regulate focal adhesion by combining single-cell RNA sequencing and reconfirmed it from bulk RNA of HCC and HCC TKIs. Drug sensitivity analysis showed that high expression of ARHGAP12 was associated with TKIs resistance, but not with immunotherapy.