Lectin affinity chromatography and quantitative proteomic analysis reveal that galectin-3 is associated with metastasis in nasopharyngeal carcinoma

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.

Hydrophilic magnetic covalent triazine frameworks for differential N-glycopeptides enrichment in breast cancer plasma membranes

“…The galectin-3-binding protein (LGALS3BP, FC SK-BR-3/MCF-10A = 18.7637) was associated with metastasis in some cancers. 40,41 These results verified the effective enrichment of the Fe 3 O 4 @CTF-PA nanoparticles. At the same time, we have conducted a gene ontology (GO) analysis on these significant up and down N-glycosites through a database search, further explored the biological information of N-glycoproteins attributable to differential N-glycosites, sorted them by significance, and selected the top 5 for analysis.…”

Clinical Assay for the Early Detection of Colorectal Cancer Using Mass Spectrometric Wheat Germ Agglutinin Multiple Reaction Monitoring

“…For instance, many cancer biomarkers in the clinic are glycoproteins, including CEA in CRC [ 6 ], carbohydrate antigen 19-9 (CA19-9) in gastrointestinal cancer [ 23 ], and prostate-specific antigen (PSA) in prostate cancer [ 24 ]. Lectin chromatography has been widely used to capture glycoproteins for MS analyses [ 25 , 26 ]. Lectins are a group of proteins that have unique affinities to carbohydrates; they can reversibly and specifically interact with certain glycan structural motifs [ 27 ].…”

Distinct cholangiocarcinoma cell migration in 2D monolayer and 3D spheroid culture based on galectin-3 expression and localization

“…The galectin-3 shRNA plasmid (sc-155994-SH) containing a pool of three to five lentiviral vector plasmids each encoding galectin-3-specific 19-25 nucleotides (plus hairpin) shRNAs, and scramble shRNA plasmid-A (sc-108060) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). The knockdown experiment was slightly modified from our previous study (39). Briefly, galectin-3 shRNA or scramble plasmids were mixed with shRNA transfection reagent (sc-108061) and shRNA transfection media (sc-108062).…”