Expression of matrix metalloproteinases (MMP-2 and MMP-9) in vocal fold polyps

Objective. Vocal fold polyps are the most common benign laryngeal lesions. Matrix metalloproteinases (MMP) play an important role in the physiological and pathological remodeling of tissues. The most important subgroup of MMP family consists of gelatinases A and B (MMP-2 and MMP-9). The objective of this study was investigation of the expression of MMP-2 and MMP-9 in vocal fold polyps and normal tissue of vocal folds. Material and methods. The immunohistochemical expression of MMP-2 and MMP-9 was investigated in specimens taken by endolaryngeal microsurgery from vocal fold polyps (n=30) and normal tissue of vocal fold (n=13, control group). Expression of MMP-2 and MMP-9, both in epithelium and stroma cells, was graded on a semiquantitative scale, ranging from 0 (no expression) to 6 points (high expression). Results. A statistically significant increase was observed in the expression of MMP-2 in stroma cells (P=0.0176) of vocal fold polyps compared to control vocal fold group, whereas no significant difference in the expression of MMP-2 was found in epithelium cells (P=0.1487). Comparison of expression of MMP-2 and MMP-9 in epithelium cells revealed a statistically significant increase in MMP-9 expression (P<0.01) in both groups. However, there was no statistically significant difference in the expression of MMP-9 between groups of vocal fold polyps and control vocal folds. Conclusion. Expression of MMP-2 in stroma was significantly higher in polyps than in normal tissue of vocal folds. Our data draw attention to the role of MMP-2 in the development of vocal fold polyps and necessity of further investigations to define its function in morphogenesis of laryngeal benign, premalignant, and malignant lesions

Matrix metalloproteinases (MMP-2,-3,-9) gene polymorphisms in cases of benign vocal fold lesions and laryngeal carcinoma

BACKGROUND/AIM: The matrix metalloproteinases (MMP) play an important role in the physiological and pathological remodeling of tissues including carcinogenesis. The study's aim was to assess the relations between MMP-2(-735C/T), MMP-2(-1306C/T), MMP-9(-1562C/T), and MMP-3(-11715A/6A) polymorphisms, and clinical/morphological manifestation of laryngeal squamous cell carcinoma (LSCC) and benign vocal fold lesions (BVFL). PATIENTS AND METHODS: Two hundred and seventeen patients with LSCC and BVFL and 458 controls were included in this study. The genotyping was performed using the real-time polymerase chain reaction method. RESULTS: The MMP-2(-1306C/T) C/T genotype was significantly rarer among the patients with moderate-poorly differentiated LSCC compared to the control group, however the MMP-3(-11715A/6A) 6A/6A genotype was significantly more frequent compared to controls. Smoking and 6A/6A genotype of MMP-3(-11715A/6A) polymorphism were associated with increased odds of LSCC risk. No associations between MMP genotypes and BVFL were found. CONCLUSION: Smoking and MMP-3 (-11715A/6A) 6A/6A genotype may cause a higher risk for developing LSCC

Association of Relative Leucocyte Telomere Length and Gene Single Nucleotide Polymorphisms ( TERT, TRF1, TNKS2) in Laryngeal Squamous Cell Carcinoma

Background/Aim: The study aimed to evaluate associations of relative leukocyte telomere length (LTL) and polymorphisms of telomere length-associated genes TERT (rs2736098), TERT-CLPTM1L (rs401681), TRF1 (rs1545827, rs10107605) and TNKS2 (rs10509637, rs10509639) in patients with laryngeal squamous cell carcinoma (LSCC). Materials and Methods: The study consisted of 300 patients with LSCC and 369 healthy control subjects. Genotyping and relative LTL measuring were carried out using qPCR. Results: Relative LTL was statistically significantly shorter in the G3 (tumor differentiation grade) subgroup of patients with LSCC compared to the G1 and G2 subgroups. Significant differences were found in genotype distributions of TERT rs401681 and TNKS2 rs10509639 between the study groups. TERT rs401681 C/T and T/T genotypes were associated with approximately 30% decreased odds of LSCC development. Conclusion: LTL was shorter in the G3 subgroup compared to the G2 and G1 subgroups of LSCC patients. TERT rs401681 and its C/T and T/T genotypes were associated with decreased odds of overall LSCC development

Determination of SIRT1 rs12778366, FGFR2 rs2981582, STAT3 rs744166, and RAGE rs1800625 Single Gene Polymorphisms in Patients with Laryngeal Squamous Cell Carcinoma

Purpose: To determine the frequency of the genotype of signal transducer and activator of transcription protein 3 (STAT3) rs744166, sirtuin (SIRT1) rs12778366, fibroblast growth factor (FGFR2) rs2981582, and advanced glycosylation end product-specific receptor (RAGE) rs1800625 gene polymorphisms in patients with laryngeal squamous cell carcinoma (LSCC). Methods: A total of 944 subjects were evaluated, which includes 144 patients with LSCC and 800 healthy controls. The genotyping of STAT3 rs744166, SIRT1 rs12778366, FGFR2 rs2981582, and RAGE rs1800625 was carried out using the RT-PCR. Results: The analysis of STAT3 rs744166, SIRT1 rs12778366, and FGFR2 rs2981582 gene polymorphisms did not reveal any differences in genotype distribution between the patients with LSCC and the control subjects. However, statistical analysis revealed that genotypes (AA, AG, and GG) of rs1800625 in RAGE gene were distributed statistically significantly differently between patients and controls (61.1%, 30.6%, and 23.6% vs. 72.5%, 25.8%, and 1.8%, respectively; p < 0.001). Additionally, statistical significance was observed in allele distribution between these two groups, i.e., allele G at rs1800625 was more frequently observed in the patient group than in controls (23.6% vs. 14.6%; p < 0.001). Conclusion: RAGE rs1800625 gene polymorphism may play a significant role in laryngeal squamous cell carcinoma development

Cross-cultural adaptation and validation of Lithuanian-NOSE scale

Purpose: To evaluate validity and reliability of Lithuanian version of Nasal Obstruction Symptom Evaluation Scale (L-NOSE), designed for the assessment of nasal obstruction. Methods: Cross-cultural adaptation of L-NOSE was accomplished according to generally accepted methodology. L- NOSE was tested for its reliability, validity, and responsiveness in the group of 50 septoplasty patients and 100 healthy volunteers' controls. Results: L- NOSE showed good internal consistency (Cronbach's alpha coefficient 0.796 for test, 0.791 for retest, 0.792 for post-operative group, and 0.817 for control group) scores and high test-retest reliability (r = 0.94, p < 0.01) scores. In patients' group, positive moderate correlations between L-NOSE scores and Sino-nasal Outcome Test-22 logically similar domain scores were found, thus indicating good convergent construct validity. L-NOSE scores for control subjects were generally lower than for patients with nasal obstruction (p < 0.001), thereby indicating good discriminant validity of questionnaire. The exploratory factor analysis confirmed one-factor structure of questionnaire. The component matrix of L-NOSE ranged from 0.667 to 0.781 (KMO = 0.754, p < 0.0001). The mean L-NOSE score improved from 58.4 ± 18.2 points to 11.1 ± 9.5 points after septoplasty (p < 0.0001), indicating good responsiveness of q

Association of Leukocyte Telomere Length and Genes Involved in Its Regulation With Oral Carcinoma

Background/aim: This study aimed to determine the relationship between the relative leukocyte telomere length (RLTL) and gene polymorphisms involved in its regulation with the occurrence of oral squamous cell carcinoma (OSCC). Patients and methods: Patients with OSCC and healthy subjects were examined. Genotyping and RLTL measurement were carried out using rPCR. Results: The OSCC group had longer telomeres than controls (p=0.001). Minor allele T at TERF1rs1545827 may increase RLTL shortening (p=0.047). TNKS2rs10509639 A/G and A/G+G/G genotypes were associated with a 2.6-fold increased odd (p=0.012) and a 2.4-fold increased odd (p=0.019) of RLTL elongation compared to A/A genotype. The A/G genotype was associated with a 2.6-fold increased odd (p=0.011) compared to the A/A+G/G genotypes. Each G allele was associated with a 2.1-fold increased odd of longer RLTL (p=0.036). Conclusion: Longer telomeres were found in patients with OSCC than in controls. The TERF1 rs1545827 and the TNKS2 rs10509639 polymorphisms were associated with an increase in RLTL

Long-Distance Communication between Laryngeal Carcinoma Cells

<div><p>Tunneling nanotubes and epithelial bridges are recently discovered new forms of intercellular communication between remote cells allowing their electrical synchronization, transfer of second messengers and even membrane vesicles and organelles. In the present study, we demonstrate for the first time in primary cell cultures prepared from human laryngeal squamous cell carcinoma (LSCC) samples that these cells communicate with each other over long distances (up to 1 mm) through membranous tunneling tubes (TTs), which can be open-ended or contain functional gap junctions formed of connexin 43. We found two types of TTs, containing F-actin alone or F-actin and α-tubulin. In the LSCC cell culture, we identified 5 modes of TT formation and performed quantitative assessment of their electrical properties and permeability to fluorescent dyes of different molecular weight and charge. We show that TTs, containing F-actin and α-tubulin, transport mitochondria and accommodate small DAPI-positive vesicles suggesting possible transfer of genetic material through TTs. We confirmed this possibility by demonstrating that even TTs, containing gap junctions, were capable of transmitting double-stranded small interfering RNA. To support the idea that the phenomenon of TTs is not only typical of cell cultures, we have examined microsections of samples obtained from human LSCC tissues and identified intercellular structures similar to those found in the primary LSCC cell culture.</p></div

Formation of TT2s between LSCC cells.

<p>(A) The leading lamellipodia of cell-1 and cell-2 indicated by white arrows are involved in cell movement. Rear lamellipodia are indicated by red arrows, and one of them outgrowing from the cell-1 forms the TT2 with the cell-2. (B–D) TT2s contain both F-actin and α-tubulin. (E) Crawling endings (“paws”, indicated by yellow arrows in A and B) of the rear and secondary lamellipodia, in addition to F-actin and α-tubulin, contain Cx43 hemichannel clusters. (F–G) Top view (DIC image) and Z-X reconstruction showing the TT2 raised above the substratum.</p

Formation of TT1s between LSCC cells.

<p>(A–D) TT1s form in the process of cell division and successive dislodgment. (E–G) TT1s contain both F-actin and α-tubulin.</p

Formation of TT5s between LSCC cells.

<p>(A–C) TT5s contain F-actin but not α-tubulin. Cx43 hemichannel clusters, which can be seen inside and/or on the TT5 surface, form functional GJs at the cell border. (D) The multiple TT5s are formed between the cell-1 and the rear lamellipodium of the cell-2 (DIC image). (E) TT5s can be formed by protrusions from apposing cells, which come into contact and form functional Cx43 GJs (green). (F and G) Several TT5s between the cell-1 and the cell-2 were found raised ∼6 µm above the substratum (DIC image).</p