Search

Background/aim: Having previously initiated genome-wide expression profiling in head and neck squamous cell carcinoma (HNSCC) for regions of the tumor, the margin of surgical resecate (MSR) and normal mucosa (NM), we here proceed with respective analysis of cases after stratification according to the expression status of tenascin (Ten)., Materials and Methods: Tissue specimens of each anatomical site were analyzed by immunofluorescent detection of Ten, fibronectin (Fn) and galectin-1 (Gal-1) as well as by microarrays., Results: Histopathological examination demonstrated that Ten + Fn + Gal-1 + co-expression occurs more frequently in samples of HNSCC (55%) than in NM (9%; p<0.01). Contrary, the Ten - Fn + Gal-1 - (45%) and Ten - Fn - Gal-1 - (39%) status occurred with significantly (p<0.01) higher frequency than in HNSCC (3% and 4%, respectively). In MSRs, different immunophenotypes were distributed rather equally (Ten + Fn + Gal-1 + =24%; Ten - Fn + Gal-1 - =36%; Ten - Fn - Gal-1 - =33%), differing to the results in tumors (p<0.05). Absence/presence of Ten was used for stratification of patients into cohorts without a difference in prognosis, to comparatively examine gene-activity signatures. Microarray analysis revealed i) expression of several tumor progression-associated genes in Ten + HNSCC tumors and ii) a strong up-regulation of gene expression assigned to lipid metabolism in MSRs of Ten - tumors, while NM profiles remained similar., Conclusion: The presented data reveal marked and specific changes in tumors and MSR specimens of HNSCC without a separation based on prognosis., (Copyright© 2018, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Background: The purpose of this study was to show our comparison of the quality of voice obtained after superficial transoral endoscopic cordectomies (types I, II, and III) according to the European Laryngological Society classification versus the more extended cordectomy (types IV and V) among patients with glottic precancerous lesions or early glottic cancer., Methods: Sixty-two patients underwent vocal assessment after cordectomy (types I-V) for dysplasia, Tis, T1a, T1b, and T2 lesions., Results: Cordectomy (types I-III) had good vocal outcomes. Cordectomy (types IV-V) showed less favorable outcomes, p < .005 in common parameters of voice range profile and symmetry in stroboscopy after cordectomy (types IV-V), with significant voice handicap index. Recurrences were found in 4 patients with T1b and T2 cancer. Two cases were managed by a higher type of cordectomy, and the other 2 by total laryngectomy., Conclusion: Effect of transoral laser microsurgery on quality of voice depends on the type of cordectomy and the site of the lesion., (© 2013 Wiley Periodicals, Inc.)

With the increasing demand for noninvasive approaches in monitoring head and neck cancer, circulating nucleic acids have been shown to be a promising tool. We focused on the global transcriptome of serum samples of head and neck squamous cell carcinoma (HNSCC) patients in comparison with healthy individuals. We compared gene expression patterns of 36 samples. Twenty-four participants including 16 HNSCC patients (from 12 patients we obtained blood samples 1 year posttreatment) and 8 control subjects were recruited. The Illumina HumanWG-6 v3 Expression BeadChip was used to profile and identify the differences in serum mRNA transcriptomes. We found 159 genes to be significantly changed (Storey's P value <0.05) between normal and cancer serum specimens regardless of factors including p53 and B-cell lymphoma family members (Bcl-2, Bcl-XL). In contrast, there was no difference in gene expression between samples obtained before and after surgery in cancer patients. We suggest that microarray analysis of serum cRNA in patients with HNSCC should be suitable for refinement of early stage diagnosis of disease that can be important for development of new personalized strategies in diagnosis and treatment of tumours but is not suitable for monitoring further development of disease.

Galectins are potent effectors of cell adhesion and growth regulation. Their expression as comples network necessitates systematic study of each member of this family. Toward this aim, we here focus on the tandem-repeat-type galectin-9. Its presence is monitored in normal squamous epithelium of the head and neck, the surgical margin, and four types of squamous cell carcinoma. Lectin presence was detected in cells of the basal layer of the epithelium. All galectin-9-negative epithelia showed aberrant positivity for keratins 14 and 19. The surgical margin presented either a normal pattern of galectin-9 and keratin presence or a mosaic-like presence/absence of galectin-9 and aberrant expression of both keratins 14 and 19. All studied specimens of squamous cell carcinoma were negative for galectin-9. When biotinylated galectin-9, or its N-terminal domain, was tested, no significant tissue reactivity for both probes was observed. Neuraminidase treatment generated reactivity to the N-domain. In conclusion, galectin-9 is expressed in the majority of samples of normal epithelium, along with regular presence of keratins 14 or 19. This lectin can represent a potential marker of normality in the cases of the studied squamous cell epithelia., (© 2012 John Wiley & Sons A/S.)

Tumor stroma is an active part influencing the biological properties of malignancies via molecular cross-talk. Cancer-associated fibroblasts play a significant role in this interaction. These cells frequently express smooth muscle actin and can be classified as myofibroblasts. The adhesion/growth-regulatory lectin galectin-1 is an effector for their generation. In our study, we set the presence of smooth muscle actin-positive cancer-associated fibroblasts in relation to this endogenous lectin and an in vivo competitor (galectin-3). In squamous cell carcinomas of head and neck, upregulation of galectin-1 presence was highly significantly correlated to presence of smooth muscle actin-positive cancer-associated fibroblasts in the tumor (p = 4 × 10(-8)). To pinpoint further correlations on the molecular level, we applied microarray analyses to the transcription profiles of the corresponding tumors. Significant correlations of several transcripts were detected with the protein level of galectin-1 in the cancer-associated fibroblasts. These activated genes (MAP3K2, TRIM23, PTPLAD1, FUSIP1, SLC25A40 and SPIN1) are related to known squamous-cell-carcinoma poor-prognosis factors, NF-κB upregulation and splicing downregulation. These results provide new insights into the significance of presence of myofibroblasts in squamous cell carcinoma., (Copyright © 2012 UICC.)

Epithelial-mesenchymal interaction between stromal fibroblasts and cancer cells influences the functional properties of tumor epithelium, including the tumor progression and spread. We compared fibroblasts prepared from stroma of squamous cell carcinoma and normal dermal fibroblasts concerning their biological activity toward normal keratinocytes assessed by immunocytochemistry and profiling of gene activation for growth factors/cytokines by microarray chip technology. IGF-2 and BMP-4 were determined as candidate factors responsible for tumor-associated fibroblast activity that influences normal epithelia. This effect was confirmed by addition of recombinant IGF-2 and BMP4, respectively, to the culture medium. This hypothesis was also verified by inhibition experiments where blocking antibodies were employed in the medium conditioned by cancer-associated fibroblast. Presence of these growth factors was also detected in tumor samples.

Regulatory T cells (Treg) are important regulators of anti-cancer immune responses, and an increase in Treg frequency was observed in the blood of cancer patients. Blood samples from 112 patients with head and neck squamous cell carcinoma antigen (HNSCC) were obtained at the time of tumour diagnosis, and lymphocyte subpopulations (CD3(+); CD3(-)CD16(+)CD56(+); CD4(+); CD8(+); CD19(+); CD4(+)CD45RA(+)) with emphasis on Treg counts (CD3(+)CD4(+)CD25(+)), complete blood count and tumour markers (squamous cell carcinoma [SCC]; CEA; alpha-1-antitrypsin [AAT]; Cyfra 21-1; C-reactive protein [CRP]) were analysed. The data were grouped according to TNM classification, and their significance for the course of the disease at an interval of 1 year after the end of the therapy was determined. The percentage of CD8(+) cells increased and the CD/D8 ratio decreased with tumour grade. The ratio of B lymphocytes decreased in patients with locoregional metastases (11.25%versus 9.22%). Treg (15.2%) and CD4(+) cells (45.3%) increased, while NK cells (11.8%) decreased in HNSCC patients compared to controls (9.0%, 38.1% and 15.8%, respectively). The data obtained at time of diagnosis were used to assess the significance of tumour markers (SCC, Cyfra 21-1 and AAT) for evaluation of prognosis. The erythrocyte counts (4.64 x 10(12)/l versus 4.45 x 10(12)/l) and haemoglobin levels (14.58 g/dl versus 14.05 g/dl) decreased, while Treg counts (8.91%versus 15.70%) increased in patients with early recurrence. Our results show that examination of these parameters could be helpful for prognostication in HNSCC patients and aid improvement of treatment strategy.

Purpose: The microenvironment established by stromal cells may or may not influence phenotypic aspects of epithelial cells and may be relevant for tumor and stem cell biology. We address this issue for keratinocytes using tumor-derived stromal cells in a co-culture system., Materials and Methods: We isolated stromal cells from human squamous cell carcinoma tissue and studied their effect on phenotypic characteristics of normal human interfollicular keratinocytes in vitro., Results: Stromal fibroblasts significantly influence immuno- and lectin cytochemical properties of co-cultured normal keratinocytes. Expression of keratins 8 and 19, the nucleolar protein nucleostemin, parameters related to adhesion/growth-regulatory galectins and the epithelial-mesenchymal transition were altered. This biological activity of tumor-derived stromal cells, which did not require cell contact, appeared to be stable, because it was maintained during passaging of keratinocytes in the absence of cancer cells., Conclusions: Tumor-derived stromal fibroblasts acquire distinct properties to shape a microenvironment conducive to altering the phenotypic characteristics of normal epithelial cells in vitro.

Background: This study presents initial data on presence of nucleostemin--a nucleolar protein typical of stem cells in the normal squamous epithelium of the oropharynx and larynx - in squamous cell carcinoma originating from these epithelia., Materials and Methods: Differentiation and proliferation markers such as keratins, beta-catenin, galectin-1, and Ki-67 were studied in parallel with nucleostemin for defining cell characteristics., Results: Nucleostemin was detected in nucleoli of both proliferating basal cells and terminally differentiated suprabasal cells of normal epithelium and in tumor cells. Importantly, malignant transformation was connected with a significant enlargement of nucleostemin-positive nucleoli in these cell types., Conclusion: Detection of nucleostemin in head and neck cancer cells, together with the size of nucleoli, may be important in the evaluation of tumor differentiation and biology.

Objective: To compare post treatment quality of life (QoL) of patients treated by radiotherapy or endoscopic transoral endolaryngeal surgery using two quality of life scoring tools., Methods: From May 1998 to July 2005, 48 patients (11 women and 37 men) with early glottic cancer were treated with curative radiotherapy (18 patients) or laser cordectomy (30 patients), and retrospectively evaluated using QoL questionnaires; European Organization for Research and Treatment of Cancer (EORTC) - EORTC-QoL Core Questionnaire (QLQ-C30 version 2.0) and organ specific EORTC - QLQ, Head and Neck Module (QLQ-H&N35) at the University Hospital Motol, Czech Republic., Results: Mean follow-up was 24 months. Only patients in complete remission were enrolled in the study. The overall score calculated separately for both questionnaires was not statistically different between both groups. Statistically significant differences were found only in specific group of questions focusing on saliva production (p=0.034) and sexuality performance (p=0.002). The majority of cases treated with cordectomy were Tis lesions. In the radiotherapy group, T1 lesions predominated (p=0.0001). Patients treated with radiotherapy were significantly older than those treated with cordectomy (p=0.027), which could explain the worsened score in sexuality questions. There were no significant differences found between genders allocated either to cordectomy or radiotherapy group., Conclusion: The overall QoL did not differ between patients treated with cordectomy or radiotherapy, despite the fact that patients treated with radiotherapy had more advanced disease and were older. There was significantly worse saliva and sexuality question score in the radiotherapy group.

Cellular activities in the regulation of growth or adhesion/migration involve protein (lectin)-carbohydrate recognition at the cell surface. Members of the galectin family of endogenous lectins additionally bind distinct intracellular ligands. These interactions with protein targets explain the relevance of their nuclear and cytoplasmic presence. Expression profiling for galectins and accessible binding sites is a histochemical approach to link localization with cellular growth properties. Non-cross-reactive antibodies for the homodimeric (proto-type) galectins-1, -2 and -7 and the chimera-type galectin-3 (Gal-3) as well as the biotinylated lectins were tested. This analysis was performed with the FaDu squamous carcinoma cell line and long-term cultured human and porcine epidermal cells as models for malignant and normal cells of squamous cell epithelial origin. A set of antibodies was added for phenotypic cell characterization. Strong nuclear and cytoplasmic signals of galectins and the differential reactivity of labeled galectins support the notion of their individual properties. The length of the period of culture was effective in modulating marker expression. Cytochemical expression profiling is a prerequisite for the selection of distinct proteins for targeted modulation of gene expression as a step toward functional analysis.

This study represents a combined lectin and immuno-histochemical analysis of normal, dysplastic and malignant squamous epithelia of the upper aerodigestive tract with emphasis on the relation between lectin reactivity and the cells' proliferative/anti-apoptotic potential. Staining by double-labeling to detect pKi67, DeltaNp63alpha, alpha2,3/6-linked sialic acid (NeuNAc) and galectin-3-reactive epitopes was performed on specimens of 8 normal cases, 27 primary tumours and 15 regional lymph node metastases. Normal epithelia expressed alpha2,6-linked NeuNAc in the basal layer, while alpha2,3-linked NeuNAc was observed in suprabasal layers. In normal epithelium pKi67 and DeltaNp63alpha positivity was seen in the basal layer and galectin-3-reactive sites in suprabasal layers. The studied squamous cell carcinomas and their metastases showed a tendency for stratification but with markedly altered architecture. The expression of the studied markers was heterogeneous between the different cancer cases but comparable between the corresponding primary and secondary lesions. Glycophenotypic properties were correlated with the level of differentiation. Tumour cell populations were characterized by occurrence of the p63+/pKi67+ alpha2,3-NeuNAc+ alpha2,6-NeuNAc+ phenotype. Analysis of the expression patterns of pKi67, p63 and galectin-3-reactive epitopes (Gal-3-RE) delineated statistically significant interrelationships (Gal-3-RE vs. p63: r = -0.709; Gal-3-RE vs. pKi67: r = -0.623; pKi67 vs. p63: r = 0.895). Of the studied markers only Gal-3-RE expression was correlated to the differentiation-dependent grading (G1 vs. G2: p = 0.007, G2 vs. G3: p = 0.006, G1 vs. G3: p = 0.002). DeltaNp63alpha expression was statistically different only between G1 and G3 (p = 0.03). No statistically significant differences were detected between the primary tumours and the corresponding regional lymph node metastases. Based on the concept of the sugar code further analysis of cell characteristics such as proliferation together with lectin histochemical features, especially using tissue lectins as probes, is thus warranted.

An impressive variety of regulatory processes including cell adhesion and migration, proliferation, apoptosis and differentiation folding and routing of glycoproteins have been found to be mediated by specific lectin-carbohydrate interactions. This article summarizes the data on glycobiological aspects of differentiation of squamous epithelia in the head and neck region under physiological conditions and in cancer. The possible function of lectins in tumor development and invasiveness is debated. Introduction of labeled endogenous lectins as a tool for the study of functional glycomics at the cellular level in head and neck squamous epithelia and carcinomas enables a complex interpretation of studied data because these lectins are normally occurring in these tissues. The lectinology of Langerhans cells in head and neck squamous epithelia and carcinoma is also mentioned. Finally, the use of the described data in the diagnosis and prospectively in the treatment of head and neck squamous cell carcinoma is shown., (Copyright 2005 S. Karger AG, Basel.)

Galectin-3, is a multifunctional effector. It is the only chimera-type member of the galectin family of endogenous lectins, which share specificity with beta-galactosides and have a jelly-roll-like folding pattern. It's activity profile includes modulation of cell-cell and cell-extracellular matrix interactions and the regulation of proliferation and apoptosis/anoikis. While lectin histochemistry with plant/invertebrate proteins is routine practice and immunohistochemical analysis of endogenous lectins has been thoroughly examined, the application of an endogenous lectin as a marker is presently primarily a promising concept. The aims of our study were to test galectin-3 as a technical probe and to correlate staining by the tissue lectin, localising accessible ligands in situ, to clinicopathological characteristics and the prognosis of patients (relapse-free and overall survival) in advanced head and neck squamous cell cancer. We measured galectin-3-dependent staining in 53 surgically resected oropharyngeal and laryngeal cancer specimens (stage III or IV). Patients were divided into two groups based on a threshold of 5% positivity in the tumour cell population. The patient's degree of positivity was significantly correlated with their level of differentiation and keratinisation and lack of lymph node involvement (P=0.0001, P=0.0007 and P=0.0224, respectively). Periods of relapse-free and overall survival were significantly shortened when the tumour population failed to meet the positivity criterion, i.e. to harbour ligands for the endogenous lectin (P=0.0039 and P=0.0259, respectively). We conclude that (a) studies with an endogenous lectin as a marker are technically feasible and (b) detection of accessible galectin-3-specific ligands is an independent prognostic marker in advanced head and neck squamous cell cancer with therapeutic potential. Of note, histochemical application of an endogenous effector after its purification and labelling may bear relevance beyond the galectins.

Head and neck squamous cell cancer (HNSCC) requires precise characterization of their biological properties to better stratify treatment approaches. Some biological features of tumor cells are related to altered glycosylation of their surface. This study characterized alpha2,3/6-N-acetylneuraminic acid (NeuNAc) expression in adult squamous epithelia of primary and metastatic HNSCC in relation to expression of well established differentiation markers, such as cytokeratins. The expression of NeuNAc was assessed by plant lectins: Maackia amurensis agglutinin type 2 (MAA) and Sambucus nigra agglutinin (SNA) specific for alpha2,3NeuNAc and alpha2,6NeuNAc, respectively. Double labeling studies at the single-cell level were performed to compare alpha2,3/6NeuNAc linkage with expression of intermediate filaments. In studied normal adult epithelia, predominantly basal expression of alpha2,6NeuNAc and suprabasal expression of alpha2,3NeuNAc was seen, showing their differentiation-dependent linkage. The cells with markers of terminal differentiation (i.e. CK10+ alpha2,3NeuNAc+ alpha2,6NeuNAc-) prevailed in HNSCC, with increasing proportion from differentiation grade G1 to G3. High proportions of cytokeratin pCK37+ carcinoma cells occurred in all studied tumors and were accompanied by a hypersialylated phenotype (i.e. alpha2,3 NeuNAc+ alpha2,6NeuNAc+) not generally seen in non-transformed epithelia. It is hypothesized that these cells could be the pool for tumor spreading in the organism. Monitoring HNSCC using multiparameter phenotype analysis can produce new data important for the understanding of the biological behavior of HNSCC, and useful for the treatment rationalization.