title: The extractability of extracellular matrix components as a marker of cartilage remodeling in laryngeal squamous cell carcinoma.
abstract: Sequential extraction was applied to investigate the proteoglycan (PG) organization in healthy laryngeal cartilage (HLC) and laryngeal cartilage squamous cell carcinoma (LCSCC). Highly stable aggrecan aggregates, extracted from both HLC and LCSCC with strong dissociative reagents, i.e., 4 M guanidine HCl (GdnHCl), represented 53% and 7%, respectively, of total extracted macromolecules. Less stable complexes/aggregates, extracted with mild dissociative reagents (1 and 2 M GdnHCl), represented 40% and 61% of total extracted PGs from healthy and cancerous cartilage, respectively. Interestingly, a relative high proportion (32%) of uronic acid (UA)-containing macromolecules were removed from the cancerous cartilage using associative extracting solutions (PBS and 0.5 M GdnHCl), which obviously represented molecules freely extractable from the tissue. In contrast, the corresponding proportion in HLC was impressively low (about 7%). The major proportion of these molecules was chondroitin sulfate-containing PGs (CSPGs), which identified mainly as aggrecan. Differential digestion of the sequential extracts with chondroitinase ABC and chondroitinase AC II demonstrated the presence of dermatan sulfate-containing PGs (DSPGs) in both HLC and LCSCC, being mainly present in the 1 M GdnHCl extract, and identified as decorin. All cancerous extracts were found to be rich in 4-sulfated disaccharides, mostly participating in DS structures. In conclusion, the applied procedure permitted the elucidation of the changes in the cartilage status, regarding the stability and identity of its proteoglycan aggregates/complexes, in both HLC and LCSCC.
authors.affiliation: Laboratory of Biochemistry, Section of Organic Chemistry, Biochemistry and Natural Products, Department of Chemistry, University of Patras, 265 00 Patras, Greece.
authors.forename: Spyros S
authors.initials: SS
authors.lastname: Skandalis
authors.forename: Dimitrios A
authors.initials: DA
authors.lastname: Theocharis
authors.forename: Nickoletta
authors.initials: N
authors.lastname: Papageorgakopoulou
authors.forename: Demitrios H
authors.initials: DH
authors.lastname: Vynios
mesh_terms: D000328:Adult; D053674:Aggrecans; D015415:Biomarkers; D002294:Carcinoma, Squamous Cell / Q000378:metabolism*; D002356:Cartilage / Q000378:metabolism*; D011508:Chondroitin Sulfate Proteoglycans / Q000032:analysis; D058575:Decorin; D005109:Extracellular Matrix / Q000737:chemistry*; D016326:Extracellular Matrix Proteins / Q000032:analysis; D006025:Glycosaminoglycans / Q000032:analysis; D006801:Humans; D007822:Laryngeal Neoplasms / Q000378:metabolism*; D037181:Lectins, C-Type; D008875:Middle Aged; D011509:Proteoglycans / Q000032:analysis
chemical_list: D053674:Aggrecans; D015415:Biomarkers; D011508:Chondroitin Sulfate Proteoglycans; C546878:DCN protein, human; D058575:Decorin; D016326:Extracellular Matrix Proteins; D006025:Glycosaminoglycans; D037181:Lectins, C-Type; D011509:Proteoglycans