Abstract
Nasopharyngeal carcinoma (NPC) is a serious cancer in East and Southeast Asia. Patients are often diagnosed at advanced stages, rendering treatment failure due to high potential of metastasis. This study identified lectin-binding glycoproteins with a potential role in NPC metastasis. Cell lysate and culture medium in highly metastatic 5-8F, and lowly-metastatic 6-10B NPC cell lines were fractionated by ConA- and WGA-affinity chromatography, and subjected to GeLC-MS/MS. A total of 232 and 197 proteins were identified in ConA-enriched fraction of 5-8F and 6-10B cell lysates respectively. In WGA-enriched fraction, 65 and 164 proteins were found in 5-8F and 6-10B cell lysates respectively. Proteins identified in culture medium for both cell lines were 223 and 85 for ConA-enriched fraction, and 94 and 124 for WGA-enriched fraction from 5-8F and 6-10B respectively. Differentially expressed proteins were functionally categorized into cell–cell adhesion, extracellular matrix, glycolysis, protein homeostasis and/or glycosylation enzymes, and lipid metabolism. Interestingly, Galectin-3 (Gal-3) was highly expressed in 5-8F cells but was lowly expressed in 6-10B cells. The Gal-3 knockdown in 5-8F cells, Gal-3 overexpression in 6-10B cells and treatment with Gal-3 inhibitor revealed that Gal-3 was responsible for metastatic phenotypes including adhesion, migration and invasion. So Galectin-3 may serve as a potential target for NPC therapeutic interventions.
Highlights
Nasopharyngeal carcinoma (NPC) is a serious cancer in East and Southeast Asia
5-8F cells exhibited lower adhesive behaviors to adhere to the extracellular matrix (ECM) (Fig. 1B), but higher migrative and invasive capabilities compared to 6-10B cells. (Fig. 1C–F)
The results demonstrated that the gal-3 knockdown 5-8F cells exhibited higher ability to attach on a monolayer of an extracellular matrix compared to the control cells, while the Galectin-3 overexpressing 6-10B cells yielded the lower adhesive index compared to the 6-10B cells harboring the control plasmid
Summary
Nasopharyngeal carcinoma (NPC) is a serious cancer in East and Southeast Asia. Patients are often diagnosed at advanced stages, rendering treatment failure due to high potential of metastasis. Cell lysate and culture medium in highly metastatic 5-8F, and lowly-metastatic 6-10B NPC cell lines were fractionated by ConA- and WGA-affinity chromatography, and subjected to GeLC-MS/MS. Proteins identified in culture medium for both cell lines were 223 and 85 for ConA-enriched fraction, and 94 and 124 for WGA-enriched fraction from 5-8F and 6-10B respectively. Proteomic data on NPC biomarkers revealed proteins linked to cancer progression involved in cell movement, cell cycling, transcription, regulation and apoptosis. These include cathepsin B, cathepsin C, cofilin-1, profilin-1, L-lactate dehydrogenase A chain, 14-3-3σ, heat shock cognate 71 kDa, and stathmin, which has been identified in secretory proteins of NPC cells[9]. Mining for glycosylated protein biomarkers in NPC has not been investigated
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