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Updated: Oct 31, 2025
"Cell Surface Capture" Workflow for Label-Free Quantification of the Cell Surface Proteome
Published on: March 24, 2023
Coupling of Cell Surface Biotinylation and SILAC-Based Quantitative Proteomics Identified Myoferlin as a Potential
Maoyu Li1,2, Fang Peng2, Guoqiang Wang2
1Department of Gastroenterology, Xiangya Hospital, Central South University, Changsha, China.
Abstract:
Distant metastasis is a major cause of treatment failure in nasopharyngeal carcinoma (NPC) patients. Cell surface proteins represent attractive targets for cancer diagnosis or therapy. However, the cell surface proteins associated with NPC metastasis are poorly understood. To identify potential therapeutic targets for NPC metastasis, we isolated cell surface proteins from two isogenic NPC cell lines, 6-10B (low metastatic) and 5-8F (highly metastatic), through cell surface biotinylation. Stable isotope labeling by amino acids in cell culture (SILAC) based proteomics was applied to comprehensively characterize the cell surface proteins related with the metastatic phenotype. We identified 294 differentially expressed cell surface proteins, including the most upregulated protein myoferlin (MYOF), two receptor tyrosine kinases(RTKs) epidermal growth factor receptor (EGFR) and ephrin type-A receptor 2 (EPHA2) and several integrin family molecules. These differentially expressed proteins are enriched in multiple biological pathways such as the FAK-PI3K-mTOR pathway, focal adhesions, and integrin-mediated cell adhesion. The knockdown of MYOF effectively suppresses the proliferation, migration and invasion of NPC cells. Immunohistochemistry analysis also showed that MYOF is associated with NPC metastasis. We experimentally confirmed, for the first time, that MYOF can interact with EGFR and EPHA2. Moreover, MYOF knockdown could influence not only EGFR activity and its downstream epithelial-mesenchymal transition (EMT), but also EPHA2 ligand-independent activity. These findings suggest that MYOF might be an attractive potential therapeutic target that has double effects of simultaneously influencing EGFR and EPHA2 signaling pathway. In conclusion, this is the first study to profile the cell surface proteins associated with NPC metastasis and provide valuable resource for future researches.
Insights
Researchers identified myoferlin (MYOF) as a key cell surface protein driving nasopharyngeal carcinoma (NPC) metastasis. MYOF targets EGFR and EPHA2 pathways, offering a potential therapeutic strategy for advanced NPC.
Area of Science:
- Oncology
- Proteomics
- Molecular Biology
Background:
- Distant metastasis is a primary reason for treatment failure in nasopharyngeal carcinoma (NPC).
- Understanding cell surface proteins linked to NPC metastasis is crucial for developing targeted therapies.
- Limited knowledge exists regarding cell surface protein involvement in NPC progression.
Purpose of the Study:
- To identify and characterize cell surface proteins associated with NPC metastasis.
- To explore the therapeutic potential of identified proteins, particularly myoferlin (MYOF).
- To elucidate the molecular mechanisms underlying MYOF's role in NPC metastasis.
Main Methods:
- Cell surface protein isolation from isogenic NPC cell lines (6-10B and 5-8F) using cell surface biotinylation.
- Proteomic analysis employing stable isotope labeling by amino acids in cell culture (SILAC) to compare protein expression.
- Validation of MYOF's role through gene knockdown experiments and immunohistochemistry.
Main Results:
- Identified 294 differentially expressed cell surface proteins, with myoferlin (MYOF) being significantly upregulated.
- MYOF knockdown suppressed NPC cell proliferation, migration, and invasion.
- Confirmed MYOF interaction with epidermal growth factor receptor (EGFR) and ephrin type-A receptor 2 (EPHA2), influencing their signaling pathways.
Conclusions:
- MYOF is a critical cell surface protein implicated in NPC metastasis.
- MYOF represents a promising therapeutic target for NPC, potentially by modulating EGFR and EPHA2 signaling.
- This study provides a valuable resource for future research into NPC metastasis and targeted therapies.
