1. Academic Validation
  2. Label-Free Quantitative Proteomics Combined with Biological Validation Reveals Activation of Wnt/β-Catenin Pathway Contributing to Trastuzumab Resistance in Gastric Cancer

Label-Free Quantitative Proteomics Combined with Biological Validation Reveals Activation of Wnt/β-Catenin Pathway Contributing to Trastuzumab Resistance in Gastric Cancer

  • Int J Mol Sci. 2018 Jul 6;19(7):1981. doi: 10.3390/ijms19071981.
Wenhu Liu 1 2 Jiangbei Yuan 3 Zhenzhong Liu 4 Jianwu Zhang 5 Jinxia Chang 6
Affiliations

Affiliations

  • 1 Department of Medicinal Chemistry, Department of Pharmacology, School of Pharmacy, North Sichuan Medical College, Nanchong 637100, China. wenhuliu@cqu.edu.cn.
  • 2 Innovative Platform of Basic Medical Sciences, Department of Microbiology and Immunology, School of Basic Medical Sciences, North Sichuan Medical College, Nanchong 637100, China. wenhuliu@cqu.edu.cn.
  • 3 School of Pharmaceutical Sciences and Innovative Drug Research Center, Chongqing University, Chongqing 401331, China. yuanjiangbei@cqu.edu.cn.
  • 4 Department of Preventive Medicine, North Sichuan Medical College, Nanchong 637100, China. liuzhenzhong@nsmc.edu.cn.
  • 5 Department of Medicinal Chemistry, Department of Pharmacology, School of Pharmacy, North Sichuan Medical College, Nanchong 637100, China. jianwuzhang@nsmc.edu.cn.
  • 6 Innovative Platform of Basic Medical Sciences, Department of Microbiology and Immunology, School of Basic Medical Sciences, North Sichuan Medical College, Nanchong 637100, China. jinxiachang@nsmc.edu.cn.
Abstract

Resistance to trastuzumab, which specifically target HER2-positive breast and gastric Cancer, can develop ultimately in Cancer patients. However, the underlying mechanisms of resistance in gastric Cancer have not been fully elucidated. Here, we established trastuzumab-resistant MKN45 and NCI N87 gastric Cancer sublines from their parental cells. The resistant cells exhibited characteristics of epithelial-mesenchymal transition (EMT) and acquired higher migratory and invasive capacities. To exploit the activated pathways and develop new strategies to overcome trastuzumab resistance, we investigated MKN45 and MKN45/R cells via label-free quantitative proteomics, and found pathways that were altered significantly in MKN45/R cells, with the Wnt/β-catenin pathway being the most significant. We further confirmed the activation of this pathway by detecting its key molecules in MKN45/R and NCI N87/R cells via Western blot, in which Wnt3A, FZD6, and CTNNB1 increased, whereas GSK-3β decreased, manifesting the activation of the Wnt/&-catenin pathway. Correspondingly, inhibition of Wnt/β-catenin pathway by ICG-001, a specific Wnt/&-catenin inhibitor, preferentially reduced proliferation and invasion of trastuzumab-resistant cells and reversed EMT. Concurringly, CTNNB1 knockdown in stable cell lines potently sensitized cells to trastuzumab and induced more Apoptosis. Taken together, our study demonstrates that the Wnt/β-catenin pathway mediates trastuzumab resistance, and the combination of Wnt/β-catenin inhibitors with trastuzumab may be an effective treatment option.

Keywords

EMT; Wnt/β-catenin pathway; gastric cancer; label-free quantitative proteomics; trastuzumab resistance.

Figures
Products