Abstract
Objective: Association of altered growth factor receptors-mediated intracellular pathways and biological processes associated with extracellular matrix composition and structure in laryngeal squamous cell carcinoma (LSCC) were described previously. In particular, the expression and glycosylation of important extracellular matrix molecules (ECM) such as small leucine rich proteoglycan lumican, may be generally associated with disrupted extracellular matrix integrity and inflammation processes which have a role in tumour invasiveness. In this study, the relative expression of different lumican glycoforms were evaluated in primary tumour and tumour-unaffected tissue samples of ten patients with metastatic and ten non-metastatic LSCC by Western blot and 2D immunoblot analysis. Materials and methods: Tissue samples from the primary tumours and paired adjacent non-tumour tissues were surgically resected from ten untreated LSCC patients with non-metastatic disease and ten LSCC patients with lymph nodes metastases. The relative expression of different lumican glycoforms in primary tumours and paired adjacent non-tumour tissues were evaluated by Western blot and 2D immunoblot analysis. Results: Results of Western blot analysis have revealed elevated expression of the moderately glycosylated lumican form in metastatic (p<0,05) and non-metastatic primary tumour tissues in comparison with tumour unaffected tissues. In addition, moderately glycosylated form of lumican with negatively charged oligosaccharide residues in the N-glycan molecule part was exclusively determined in metastatic primary tumour tissues by 2D immunoblot analysis. Conclusion: We demonstrated elevated expression of the moderately glycosylated lumican form with negatively charged oligosaccharide residues in the N-glycan portion exclusively in primary laryngeal squamous cell carcinoma from patients with metastatic disease.
Highlights
Laryngeal squamous cell carcinoma (LSCC) is a multifactorial disease characterized by frequent metastasis in lymph nodes early during disease onset, and represents the most common form of head and neck malignancies[1]
Recent evidence indicates a clear association between altered growth factor receptors-mediated intracellular pathways and biological processes underlying LSCC that induce changes in the extracellular matrix (ECM)[2,3]
Lumican is an important ECM protein that belongs to a class II small leucine rich proteoglycans and has four potential N-glycosylation sites[4]
Summary
Laryngeal squamous cell carcinoma (LSCC) is a multifactorial disease characterized by frequent metastasis in lymph nodes early during disease onset, and represents the most common form of head and neck malignancies[1]. Lumican is an important ECM protein that belongs to a class II small leucine rich proteoglycans and has four potential N-glycosylation sites[4]. Depending on the tissue type, it is expressed in different structural forms, including the core protein with MW approximately 40 kDa, the 70 kDa proteoglycan substituted with keratane-sulphate or nonsulphated polylactosamine chain and the 50 kDa glycoprotein form[5]. Depending on its glycosylation status, lumican bears different biological functions or contributes to progression of malignancies by stimulating cellular signalization, proliferation and migration[6,7]. We hypothesized that expression of different glycoforms of lumican might be important for the LSCC pathogenesis. We present data on lumican glycoforms expression in a set of 10 metastatic and 10 non-metastatic primary LSCC tumours
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