Pyrotinib targeted EGFR/GRP78 mediated cell apoptosis in high EGFR gene copy number gastric cancer

Abstract Background Despite frequent Epidermal Growth Factor Receptor (EGFR) amplification and overexpression in gastric cancer, limited therapeutic responses were observed in existing EGFR-targeted agents. Pyrotinib, an irreversible dual EGFR/HER2 tyrosine kinase inhibitor, has shown clinical effic...

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Main Authors: Lingbo Bao, Xudong Wang, Xiuyong Liao, Dong Li, ChunXue Li, Nan Dai, Xiaoyan Dai, Jing Yang, Nana Hu, Xueling Tong, Zhenjie He, Yuancheng Zhao, Zheng Liu, Yue Hu, Jinlu Shan, Dong Wang, Mengxia Li, Qian Chen
Format: Article
Language:English
Published: BMC 2025-08-01
Series:Journal of Experimental & Clinical Cancer Research
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Online Access:https://doi.org/10.1186/s13046-025-03485-6
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author Lingbo Bao
Xudong Wang
Xiuyong Liao
Dong Li
ChunXue Li
Nan Dai
Xiaoyan Dai
Jing Yang
Nana Hu
Xueling Tong
Zhenjie He
Yuancheng Zhao
Zheng Liu
Yue Hu
Jinlu Shan
Dong Wang
Mengxia Li
Qian Chen
author_facet Lingbo Bao
Xudong Wang
Xiuyong Liao
Dong Li
ChunXue Li
Nan Dai
Xiaoyan Dai
Jing Yang
Nana Hu
Xueling Tong
Zhenjie He
Yuancheng Zhao
Zheng Liu
Yue Hu
Jinlu Shan
Dong Wang
Mengxia Li
Qian Chen
author_sort Lingbo Bao
collection DOAJ
description Abstract Background Despite frequent Epidermal Growth Factor Receptor (EGFR) amplification and overexpression in gastric cancer, limited therapeutic responses were observed in existing EGFR-targeted agents. Pyrotinib, an irreversible dual EGFR/HER2 tyrosine kinase inhibitor, has shown clinical efficacy in HER2-driven malignancies, but its potential role in EGFR-high copy number gastric cancer remains to be investigated. Methods Using EGFR-high copy number gastric cancer cell lines, primary cells and subcutaneous tumor models in nude mice, we systematically evaluated pyrotinib’s anti-tumor activity through viability assays, apoptosis analysis, and transcriptomic profiling. Mechanistic studies included co-immunoprecipitation, proximity ligation assays, ubiquitination assays, and RNA sequencing. Results Pyrotinib selectively suppressed proliferation, induced apoptosis, and chemosensitized in EGFR-high copy number gastric cancer models. Mechanistically, pyrotinib promoted EGFR-GRP78 (Glucose-regulated protein 78) complex formation in the endoplasmic reticulum, activating the protein kinase R-like endoplasmic reticulum kinase/ activating transcription factor 4/ C-EBP homologous protein (PERK/ATF4/CHOP) axis to drive ER stress-mediated apoptosis. Concurrently, pyrotinib inhibited GRP78 phosphorylation at Thr62, triggering K48-linked ubiquitination (ubiquitin chains formed via lysine 48 linkages) and proteasomal degradation, which impaired DNA double-strand break (DSB) repair and sensitized cells to oxaliplatin-induced γ-H2A.X accumulation. Conclusion This translational study suggests that pyrotinib combined with oxaliplatin may serve as a promising strategy for patients with EGFR-high copy number gastric cancer and highlighted the discovery of this previously unknown EGFR/ GRP78 signaling axis, which provides the molecular basis and the rationale to target EGFR.
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issn 1756-9966
language English
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publisher BMC
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spelling doaj-art-a73556a5305a402a80a75200ef49e90d2025-08-24T11:57:30ZengBMCJournal of Experimental & Clinical Cancer Research1756-99662025-08-0144111810.1186/s13046-025-03485-6Pyrotinib targeted EGFR/GRP78 mediated cell apoptosis in high EGFR gene copy number gastric cancerLingbo Bao0Xudong Wang1Xiuyong Liao2Dong Li3ChunXue Li4Nan Dai5Xiaoyan Dai6Jing Yang7Nana Hu8Xueling Tong9Zhenjie He10Yuancheng Zhao11Zheng Liu12Yue Hu13Jinlu Shan14Dong Wang15Mengxia Li16Qian Chen17Cancer Center of Daping Hospital, Army Medical UniversityCancer Center of Daping Hospital, Army Medical UniversityDepartment of Oncology, Chongqing University Qianjiang Hospital, Qianjiang Central Hospital of ChongqingDepartment of Oncology, General Hospital of Western Theater CommandDepartment of General Surgery, Daping Hospital, Third Military Medical University (Army Medical University)Cancer Center of Daping Hospital, Army Medical UniversityCancer Center of Daping Hospital, Army Medical UniversityCancer Center of Daping Hospital, Army Medical UniversityCancer Center of Daping Hospital, Army Medical UniversityCancer Center of Daping Hospital, Army Medical UniversityCancer Center of Daping Hospital, Army Medical UniversityDepartment of Ophthalmology, The General Hospital of Western Theater CommandCancer Center of Daping Hospital, Army Medical UniversityCancer Center of Daping Hospital, Army Medical UniversityCancer Center of Daping Hospital, Army Medical UniversityDepartment of Oncology, Chongqing University Qianjiang Hospital, Qianjiang Central Hospital of ChongqingCancer Center of Daping Hospital, Army Medical UniversityCancer Center of Daping Hospital, Army Medical UniversityAbstract Background Despite frequent Epidermal Growth Factor Receptor (EGFR) amplification and overexpression in gastric cancer, limited therapeutic responses were observed in existing EGFR-targeted agents. Pyrotinib, an irreversible dual EGFR/HER2 tyrosine kinase inhibitor, has shown clinical efficacy in HER2-driven malignancies, but its potential role in EGFR-high copy number gastric cancer remains to be investigated. Methods Using EGFR-high copy number gastric cancer cell lines, primary cells and subcutaneous tumor models in nude mice, we systematically evaluated pyrotinib’s anti-tumor activity through viability assays, apoptosis analysis, and transcriptomic profiling. Mechanistic studies included co-immunoprecipitation, proximity ligation assays, ubiquitination assays, and RNA sequencing. Results Pyrotinib selectively suppressed proliferation, induced apoptosis, and chemosensitized in EGFR-high copy number gastric cancer models. Mechanistically, pyrotinib promoted EGFR-GRP78 (Glucose-regulated protein 78) complex formation in the endoplasmic reticulum, activating the protein kinase R-like endoplasmic reticulum kinase/ activating transcription factor 4/ C-EBP homologous protein (PERK/ATF4/CHOP) axis to drive ER stress-mediated apoptosis. Concurrently, pyrotinib inhibited GRP78 phosphorylation at Thr62, triggering K48-linked ubiquitination (ubiquitin chains formed via lysine 48 linkages) and proteasomal degradation, which impaired DNA double-strand break (DSB) repair and sensitized cells to oxaliplatin-induced γ-H2A.X accumulation. Conclusion This translational study suggests that pyrotinib combined with oxaliplatin may serve as a promising strategy for patients with EGFR-high copy number gastric cancer and highlighted the discovery of this previously unknown EGFR/ GRP78 signaling axis, which provides the molecular basis and the rationale to target EGFR.https://doi.org/10.1186/s13046-025-03485-6PyrotinibEGFRGRP78UbiquitinationTRIM21
spellingShingle Lingbo Bao
Xudong Wang
Xiuyong Liao
Dong Li
ChunXue Li
Nan Dai
Xiaoyan Dai
Jing Yang
Nana Hu
Xueling Tong
Zhenjie He
Yuancheng Zhao
Zheng Liu
Yue Hu
Jinlu Shan
Dong Wang
Mengxia Li
Qian Chen
Pyrotinib targeted EGFR/GRP78 mediated cell apoptosis in high EGFR gene copy number gastric cancer
Journal of Experimental & Clinical Cancer Research
Pyrotinib
EGFR
GRP78
Ubiquitination
TRIM21
title Pyrotinib targeted EGFR/GRP78 mediated cell apoptosis in high EGFR gene copy number gastric cancer
title_full Pyrotinib targeted EGFR/GRP78 mediated cell apoptosis in high EGFR gene copy number gastric cancer
title_fullStr Pyrotinib targeted EGFR/GRP78 mediated cell apoptosis in high EGFR gene copy number gastric cancer
title_full_unstemmed Pyrotinib targeted EGFR/GRP78 mediated cell apoptosis in high EGFR gene copy number gastric cancer
title_short Pyrotinib targeted EGFR/GRP78 mediated cell apoptosis in high EGFR gene copy number gastric cancer
title_sort pyrotinib targeted egfr grp78 mediated cell apoptosis in high egfr gene copy number gastric cancer
topic Pyrotinib
EGFR
GRP78
Ubiquitination
TRIM21
url https://doi.org/10.1186/s13046-025-03485-6
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