Coupling of Cell Surface Biotinylation and SILAC-Based Quantitative Proteomics Identified Myoferlin as a Potential Therapeutic Target for Nasopharyngeal Carcinoma Metastasis

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.

ZBTB7A governs 2-DG-inhibited glycolysis by regulating GLUT1 in nasopharyngeal carcinoma

“…ZBTB7A, or Pokemon, showed complex characteristics and was expressed at different levels in NPC cells. 5-8F and 6-10B cells were used as the research objects because of their high and low metastatic potential, respectively (Li et al, 2021). WB assays showed that ZBTB7A expression in 5-8F was higher than in 6-10B cells (Liu et al, 2019).…”

Research progress of proteomics in screening tumor markers of gastric cancer

“…Proteomics can analyze the dynamics of protein composition, expression level, and modification status in the organism from a holistic perspective by comparing different proteins, understanding protein-protein interaction (PPI) and linkage, and revealing the pattern of protein function and life activity. Essential for understanding the development, prevention, diagnosis, and treatment of cancer [2].…”

Discovery of a Highly Potent and Selective MYOF Inhibitor with Improved Water Solubility for the Treatment of Gastric Cancer

“…13,16 Additionally, overexpression of MYOF accelerates the process of epithelial−mesenchymal transition (EMT) to promote the invasion and metastasis of cancer cells, and inhibition of MYOF induces the transition of mesenchymal cells to the epithelial cells. 11,19,20 Briefly, MYOF plays a vital role in various tumors and is becoming an emerging cancer therapeutic target. Since the full-length protein crystal structure of MYOF has not been revealed yet, the studies for developing novel MYOF inhibitors are encountering more challenges.…”

“…Additionally, recent studies demonstrated that MYOF was lowly expressed in normal cells but highly expressed in various malignant tumors such as gastric cancer, , breast cancer, , colon cancer, pancreatic cancer, , B-cell lymphoma, and liver cancer . At the same time, the high expression of MYOF significantly reduced the survival time of patients and was strongly correlated with poor prognosis. , Additionally, overexpression of MYOF accelerates the process of epithelial–mesenchymal transition (EMT) to promote the invasion and metastasis of cancer cells, and inhibition of MYOF induces the transition of mesenchymal cells to the epithelial cells. ,, Briefly, MYOF plays a vital role in various tumors and is becoming an emerging cancer therapeutic target.…”