The c.*229C > T gene polymorphism in 3'UTR region of the topoisomerase IIß binding protein 1 gene and LOH in BRCA1/2 regions and their effect on the risk and progression of human laryngeal carcinoma.

Topoisomerase IIß binding protein 1 (TopBP1), a multiple-BRCT-domain, protein plays crucial roles in chromosome replication, DNA damage repair, apoptosis, and cell cycle checkpoint signalling. The aim of this study was to identify five SNPs at loci potentially located in the 3'UTR region of the TopBP1 gene (rs185903567, rs116645643, rs115160714, rs116195487, rs112843513), their relationship with the risk of squamous cell laryngeal cancer (SCLC), tumor invasiveness, and prognosis. Genotyping was performed in 323 genetically unrelated individuals with SCLC and 418 randomly selected healthy volunteers. Allele-specific TopBP1 mRNA and protein expressions were determined by using real-time PCR and Western blotting techniques, respectively. LOH in BRCA1/BRCA2 was determined by using microsatellite markers. Compared to homozygous common allele carriers, heterozygosity for the T variant was associated with increased risk of SCLC (adjusted odds ratio [OR] = 9.83, 95 % confidence interval [CI]: 3.12-22.16, p dominant < 0.0001). The presence of risk allele at rs115160714 TopBP1 determined a higher incidence of nodal metastases (OR = 7.98, 95 % CI: 3.94-16.00, p = 0.001) and higher tumor grade (OR = 6.48, 95 % CI: 0.86-48.01, p = 0.03). The heterozygotes displayed diffuse tumor growth with no distinct borderline (OR = 3.10, 95 % Cl: 0.92-10.62, p = 0.049) and higher depth of invasion (OR = 2.66, 95 % Cl: 0.78-9.03, p = 0.04). Relationships were also identified between TopBP1 mRNA/protein expression and overall survival (p < 0.0001). The incidence of LOH in BRCA1/BRCA2 was significantly related to higher tumor grade and TFG (p < 0.05). The results of this study suggest that rs115160714 TopBP1 may be a genetic marker of etiology and progression in laryngeal cancer.

Gene/protein expression of CAPN1/2-CAST system members is associated with ERK1/2 kinases activity as well as progression and clinical outcome in human laryngeal cancer.

Recent evidence indicates the involvement of calpains (CAPNs), a family of cysteine proteases, in cancer development and progression, as well as the insufficient response to cancer therapies. The contribution of CAPNs and regulatory calpastatin (CAST) and ERK1/2 kinases to aggressiveness, disease course, and outcome in laryngeal cancer remains elusive. This study was aimed to evaluate the CAPN1/2-CAST-ERK1/2 enzyme system mRNA/protein level and to investigate whether they can promote the dynamic of tumor growth and prognosis. The mRNA expression of marker genes was determined in 106 laryngeal cancer (SCLC) cases and 73 non-cancerous adjacent mucosa (NCLM) controls using quantitative real-time PCR. The level of corresponding proteins was analyzed by Western Blot. SLUG expression, as indicator of pathological advancement was determined using IHC staining. Significant increases of CAPN1/2-CAST-ERK1/2 levels of mRNA/protein were noted in SCLC compared to NCLM (p < 0.05). As a result, a higher level of CAPN1 and ERK1 genes was related to larger tumor size, more aggressive and deeper growth according to TFG scale and SLUG level (p < 0.05). There were also relationships of CAPN1/2 and ERK1 with incidences of local/nodal recurrences (p < 0.05). An inverse association for CAPN1/2, CAST, and ERK1/2 transcripts was determined with regard to overall survival (p < 0.05). In addition, a higher CAPN1 and phospho-ERK1 protein level was related to higher grade and stage (p < 0.05) and was found to promote worse prognosis. This is the first study to show that activity of CAPN1/2- CAST-ERK1/2 axis may be an indicator of tumor phenotype and unfavorable outcome in SCLC.

The effect of metallothionein 2A core promoter region single-nucleotide polymorphism on accumulation of toxic metals in sinonasal inverted papilloma tissues.

Metallothioneins (MTs) are intracellular thiol-rich heavy metal-binding proteins which join trace metal ions protecting cells against heavy metal toxicity and regulate metal distribution and donation to various enzymes and transcription factors. The goal of this study was to identify the -5 A/G (rs28366003) single-nucleotide polymorphism (SNP) in the core promoter region of the MT2A gene, and to investigate its effect on allele-specific gene expression and Cd, Zn, Cu and Ni content in sinonasal inverted papilloma tissue (IP), with non-cancerous sinonasal mucosa (NCM) as a control. The MT2A promoter region -5 A/G SNP was identified by restriction fragment length polymorphism using 117 IP and 132 NCM. MT2A gene analysis was performed by quantitative real-time PCR. Metal levels were analyzed by flame atomic absorption spectrometry. The frequency of A allele carriage was 99.2% and 100% in IP and NCM, respectively. The G allele carriage was detected in 23.9% of IP and in 12.1% of the NCM samples. As a result, a significant association of -5 A/G SNP in MT2A gene with mRNA expression in both groups was determined. A significant association was identified between the -5 A/G SNP in the MT2A gene with mRNA expression in both groups. A highly significant association was detected between the rs28366003 genotype and Cd and Zn content in IP. Furthermore, significant differences were identified between A/A and A/G genotype with regard to the type of metal contaminant. The Spearman rank correlation results showed the MT2A gene expression and both Cd and Cu levels were negatively correlated. The results obtained in this study suggest that the -5 A/G SNP in the MT2A gene may have an effect on allele-specific gene expression and toxic metal accumulation in sinonasal inverted papilloma.

Gene and protein expression of glucose transporter 1 and glucose transporter 3 in human laryngeal cancer-the relationship with regulatory hypoxia-inducible factor-1α expression, tumor invasiveness, and patient prognosis.

Increased glucose uptake mediated by glucose transporters and reliance on glycolysis are common features of malignant cells. Hypoxia-inducible factor-1α supports the adaptation of hypoxic cells by inducing genes related to glucose metabolism. The contribution of glucose transporter (GLUT) and hypoxia-inducible factor-1α (HIF-1α) activity to tumor behavior and their prognostic value in head and neck cancers remains unclear. The aim of this study was to examine the predictive value of GLUT1, GLUT3, and HIF-1α messenger RNA (mRNA)/protein expression as markers of tumor aggressiveness and prognosis in laryngeal cancer. The level of hypoxia/metabolic marker genes was determined in 106 squamous cell laryngeal cancer (SCC) and 73 noncancerous matched mucosa (NCM) controls using quantitative real-time PCR. The related protein levels were analyzed by Western blot. Positive expression of SLC2A1, SLC2A3, and HIF-1α genes was noted in 83.9, 82.1, and 71.7% of SCC specimens and in 34.4, 59.4, and 62.5% of laryngeal cancer samples. Higher levels of mRNA/protein for GLUT1 and HIF-1α were noted in SCC compared to NCM (p < 0.05). SLC2A1 was found to have a positive relationship with grade, tumor front grading (TFG) score, and depth and mode of invasion (p < 0.05). SLC2A3 was related to grade and invasion type (p < 0.05). There were also relationships of HIF-1α with pTNM, TFG scale, invasion depth and mode, tumor recurrences, and overall survival (p < 0.05). In addition, more advanced tumors were found to be more likely to demonstrate positive expression of these proteins. In conclusion, the hypoxia/metabolic markers studied could be used as molecular markers of tumor invasiveness in laryngeal cancer.

Metallothionein 2A core promoter region genetic polymorphism and its impact on the risk, tumor behavior, and recurrences of sinonasal inverted papilloma (Schneiderian papilloma).

Inverted papillomas are a unique group of locally aggressive benign epithelial neoplasms in the nasal cavity and paranasal sinuses arising from the Schneiderian mucosa. Metallothioneins are sulfhydryl-rich heavy metal-binding proteins required for metal toxicity protection and regulation of biological mechanisms including proliferation and invasion. The goal of this study was to identify three SNPs at loci -5 A/G (rs28366003) and -209 A/G (rs1610216) in the core promoter region and at locus +838 C/G (rs10636) in 3'UTR region of the MT2A gene with IP risk and with tumor invasiveness according to Krouse staging. Genotyping was performed using the PCR restriction fragment length polymorphism technique in 130 genetically unrelated IP individuals, and 418 randomly selected healthy volunteers. The presence of the rs28366003 SNP was significantly related to the risk of IP within the present population-based case-control study. Compared to homozygous common allele carriers, heterozygosity and homozygosity for the G variant had a significantly increased risk of IP (adjusted odds ratio [OR] = 7.71, 95% confidence interval [CI]: 4.01-14.91, p(dominant) < 0.001). Moreover, risk allele carriers demonstrated higher Krouse stage (pT1 vs. pT2-4) (OR = 19.32; 95% CI, 2.30-173.53; p < 0.0001), diffuse tumor growth (OR = 4.58; 95% CI, 1.70-12.11; p = 0.0008), bone destruction (OR = 4.13; 95% CI, 1.50-11.60; p = 0.003), and higher incidence of tumor recurrences (OR = 5.11; 95% CI, 1.68-15.20; p = 0.001). The findings suggest that MT2A gene variation rs28366003 may be implicated in the etiology of sinonasal inverted papilloma in a Polish population.

Expression of Th17 cell population regulatory cytokines in laryngeal carcinoma - Preliminary study.

AIM OF THE STUDY: Aim of the study was to evaluate the potential role of regulatory and proinflammatory cytokines IL-23 and IL-17 as Th17 lymphocyte activity markers in relation to invasiveness in laryngeal cancer. MATERIAL AND METHODS: The immunological analysis was conducted in 50 patients treated for squamous cell laryngeal carcinoma and 30 healthy volunteers as controls. The levels of IL-23 and IL-17 in supernatants of purified peripheral blood mononuclear cell cultures were determined by using the enzyme-linked immunosorbent assay (ELISA). The clinicomorphological criteria included pTNM, stage, G, and the total tumour front grading (TFG) score. RESULTS: Our data demonstrated higher concentrations of IL-23 in patients as compared to controls (p = 0.0001). No statistical difference for IL-17 in these groups was observed. Our study revealed significant dependences in IL-23 expression on pT (p = 0.04), histological differentiation (p = 0.04), and TFG total score (p = 0.02). Advanced tumours (pT3-pT4) with higher grade (G2-G3) and higher invasiveness (> 14 TFG points) were characterised by elevated IL-23 levels in PBMC supernatants. Our data did not indicate a relationship between cytokine levels and three- and five-year survival. However, a tendency towards lower content of IL-23 in PBMC cultures in patients who lived longer than five years after treatment was noted. The relationships between IL-17 level in PBMC cultures and clinicomorphological and prognostic parameters have not been disclosed. CONCLUSIONS: The results of this study suggest the importance of regulatory cytokine IL-23 in determining the aggressive potential of laryngeal carcinomas.

Expression of prostaglandin E2 prostanoid receptor EP2 and interleukin-1ß in laryngeal carcinoma - preliminary study.

AIM OF THE STUDY: Expression of EP2 protein, the prostaglandin E2 (PGE2) receptor, produced by tumour microenvironment inflammatory cells as well as tumour cells, may promote cellular proliferation and growth in an autocrine and paracrine fashion. The phenomenon involving these proteins is regulated by interleukin 1ß (IL-1ß). Many researchers indicate a connection of EP2 and IL-1ß in various types of neoplasms with higher tumour progression and poor prognosis. The aim of this study was to analyse the EP2 expression within laryngeal carcinoma tissue and IL-1ß levels in peripheral blood mononuclear cell supernatants and to find relationships between clinicomorphological features. MATERIAL AND METHODS: A group of 50 patients with verified squamous cell laryngeal carcinoma was analysed in this study. The pathological evaluation included pTNM depth of invasion according to tumour front grading criteria. Immunohistochemical analysis for membranous staining of EP2 in tumour tissues was used. The IL-1ß expression was determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: Increased EP2 expression in carcinoma cells was confirmed for more advanced tumours (pT3-pT4 vs. pT1-pT2, p < 0.0001 and pN1-3 vs. pN0, p = 0.02). Tumours with the highest aggressiveness identified by deeper invasion of submucosa or cartilage were characterised by the highest expression of EP2 (p < 0.0001). In laryngeal carcinomas characterised by a lower differentiation the highest EP2 expression in tumour cells was noted (p = 0.009). A positive relationship between IL-1ß expression and the presence of lymph node metastases was also confirmed (p = 0.04). CONCLUSIONS: The study indicates the potential effect of EP2 receptor and IL-1ß on tumour progression in laryngeal carcinoma.

The -5 A/G single-nucleotide polymorphism in the core promoter region of MT2A and its effect on allele-specific gene expression and Cd, Zn and Cu levels in laryngeal cancer.

Metallothioneins (MTs) are low molecular weight, cysteine-rich heavy metal-binding proteins which participate in the mechanisms of Zn homeostasis, and protect against toxic metals. MTs contain metal-thiolate cluster groups and suppress metal toxicity by binding to them. The aim of this study was to determine the -5 A/G (rs28366003) single-nucleotide polymorphism (SNP) in the core promoter region of the MT2A gene and to investigate its effect on allele-specific gene expression and Cd, Zn and Cu content in squamous cell laryngeal cancer (SCC) and non-cancerous laryngeal mucosa (NCM) as a control. The MT2A promoter region -5 A/G SNP was determined by restriction fragment length polymorphism using 323 SCC and 116 NCM. MT2A gene analysis was performed by quantitative real-time PCR. The frequency of A allele carriage was 94.2% and 91.8% in SCC and NCM, respectively, while G allele carriage was detected in 5.8% and 8.2% of SCC and NCM samples, respectively. As a result, a significant association was identified between the -5 A/G SNP in the MT2A gene with mRNA expression in both groups. Metal levels were analyzed by flame atomic absorption spectrometry. The significant differences were identified between A/A and both the A/G and G/G genotypes, with regard to the concentration of the contaminating metal. The Spearman rank correlation results showed that the MT2A expression and Cd, Zn, Cu levels were negatively correlated. Results obtained in this study suggest that -5 A/G SNP in MT2A gene may have an effect on allele-specific gene expression and accumulation of metal levels in laryngeal cancer.

Expression of cell adhesion molecules in laryngeal carcinoma - preliminary analysis.

AIM OF THE STUDY: Intercellular adhesion molecules present in immunocompetent cells as well as endothelium and tumour cells can regulate cell migration, angiogenesis, apoptosis, proliferation, and metastases in solid tumours. The aim of this study was to analyse the sICAM-1 (soluble intercellular adhesion molecule 1) and sVCAM-1 (soluble vascular cell adhesion molecule 1) expression in peripheral blood mononuclear cell (PBMC) cultures, and to find their relationships with clinicomorphological characteristics in laryngeal cancer. MATERIALS AND METHODS: The analysis included a group of 50 patients with verified squamous cell carcinoma of the larynx. The control group constituted 30 healthy volunteers. The pathological assessment included pTNM, stage, histological grade, and type of invasion according to the tumour front grading. The expression of adhesion molecules was assessed using the enzyme-linked immunosorbent assay (ELISA). RESULTS: Increased expression of sICAM-1 and sVCAM-1 was an indicator of more aggressive laryngeal carcinomas. More advanced local changes evaluated on the pT feature were connected with a higher sVCAM-1 (p = 0.017), but not sICAM-1 level. The presence of lymph node metastases correlated with a higher expression of adhesion molecules (p = 0.012 and p = 0.003, for sICAM-1 and sVCAM-1, respectively). Tumours with more diffuse growth and infiltrating with small cell groups (< 15/hpf) was characterised by the highest level of adhesive proteins (p = 0.001 and p = 0.02 for sICAM and sVCAM, respectively). Moreover, lower levels of sICAM-1 and sVCAM-1 were observed more frequently in patients who lived longer than five years after treatment. CONCLUSIONS: The study indicates the importance of the sICAM and sVCAM expression as indicators of advanced changes and prognosis in patients with laryngeal carcinoma.

Diagnostic impact of promoter methylation and E-cadherin gene and protein expression levels in laryngeal carcinoma.

AIM OF THE STUDY: Inactivation of the tumor suppressor E-cadherin (CDH1) and its decreased expression is an important occurrence during carcinogenesis. Nevertheless, the relationship of CDH1 expression and the promoter methylation with laryngeal cancer cell aggressiveness is still unclear. The purpose of this study was to elucidate the gene and protein E-cadherin expression and the DNA methylation levels and to describe the correlations with morphological features in squamous cell laryngeal cancer. MATERIAL AND METHODS: The authors studied E-cadherin and the DNA methylation level in 86 cases to gain a further understanding of the clinicopathologic significance of analyzed parameters. The pathological evaluation included pTNM classification and the tumor front grading (TFG) criteria. Quantitative analysis of the amplified product in real time (qRT-PCR) for estimation of CDH1 mRNA was used. The methylation status was investigated by using methyl-specific polymerase chain reaction (MSP). The level of CDH1 protein expression by Western blot was determined. RESULTS: Downregulation of E-cadherin was found to be related to promoter methylation (p < 0.001). In tumors with the highest invasiveness according to TFG criteria the lowest E-cadherin gene and protein level in the study group was observed (p = 0.046 and p = 0.0002, respectively). In SCLC with muscle and cartilage invasion and disperse infiltration the lowest CDH1 gene and protein expression was noted (p = 0.0003 and p = 0.003 for deep invasion, p = 0.033 and p = 0.003 for multifocal infiltration, respectively). CONCLUSIONS: The current findings suggest an association of E-cadherin tumor expression with progression of laryngeal cancer. CDH1 gene level may be an auxiliary molecular marker for advanced cases of laryngeal carcinoma; however, further studies are necessary.

Expression of CTLA-4 and Foxp3 in peripheral blood T cells of patients with squamous cell laryngeal carcinoma.

Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4, CD152) and Foxp3 (forkhead box P3) are receptors present on T cells which play a critical role in the down-regulation of antigen-activated immune responses. To evaluate the potential influences of CTLA-4 and Foxp3 on cancer invasiveness, a case-control study was conducted in 86 patients treated for squamous cell laryngeal carcinoma. The abundance of CTLA-4 and Foxp3 gene transcripts in the purified peripheral blood mononuclear cells (PBMCs) by quantitative real-time PCR (qRT-PCR) was determined. The analysis of proteins by Western blot was performed. The relationships between CTLA-4 and Foxp3 gene and protein expression as well as the aggressiveness of tumor determined on pT, type and depth of invasion were investigated. Our work revealed a significant dependence of mRNA CTLA-4 on tumor front grading (TFG) total score (p = 0.04) as well as CTLA-4 protein expression on pT (p = = 0.03) and type of invasion (p = 0.03). Advanced pT3-pT4 tumors with diffuse infiltration and > 14 TFG points were characterized by higher average values of CTLA-4 protein in PBMCs. Our data also demonstrated significant differences between Foxp3 protein levels in relation to pT (p = 0.04), depth of invasion (p = = 0.02) and type of invasion (p = 0.03). In tumors with the highest invasiveness identified by the pT3-pT4 status, deep invasion with involvement of cartilage and diffuse infiltration, the highest Foxp3 protein level was observed. In conclusion, these results suggest an impact of CTLA-4 and Foxp3 in determining proliferative and aggressive potential of laryngeal carcinoma, highlighting the significance of CTLA-4 and Foxp3 as potential predictive indicators.

Impact of EGFR immunoexpression on STAT3 activation and association with proinflammatory/regulatory cytokine pattern in laryngeal squamous cell carcinoma.

Stimulation of epidermal growth factor receptor (EGFR) results in the activation of signal transducer and activator of transcription-3 (STAT3), a transcriptional factor associated with carcinogenesis. Proinflammatory cytokines are capable of activating a tumor cell death program by reducing EGFR tyrosine phosphorylation. This study aimed to identify EGFR expression in laryngeal carcinoma and determine the relationship with STAT3 and proinflammatory/regulatory cytokine secretion. An analysis of EGFR expression (membranous EGFR-m and cytoplasmic EGFR-c) was performed in tumor tissues by immunohistochemical (IHC) staining in 45 medical cases of laryngeal carcinoma. STAT3 expression in freshly isolated tumor and non-cancerous normal epithelial cells by RT-PCR was analyzed in 24 patients after total larynx resection. The concentrations of TNFalpha, IL-2, IL-6, IL-8 and IFNgamma secreted by purified peripheral blood mononuclear cells (PBMCs) or contained in whole blood samples were measured by ELISA. The relationship between EGFR and mRNA STAT3 expression as well as the level of secreted cytokines was investigated. In our study, 93.3% tumors expressed EGFR-m and 37.8% EGFR-c. It also revealed a statistically significant dependence of the EGFR status on STAT3 expression in neoplastic tissues. Tumors with IHC EGFR-m positive staining >50% of the total number of cells, as well as with EGFR-c positive staining, were characterized by the most frequent presence of STAT3 expression. Our data demonstrate a significant negative relationship between EGFR-m expression and TNFalpha concentration, and a positive connection between membranous EGFR and IL-8 or IFNgamma levels recorded in isolated PBMCs. Furthermore, this study revealed a significant relationship between EGFR-c immunoexpression and IL-8 or IFNgamma concentration. Our findings have confirmed a key role of EGFR in determining the proliferative and malignant potential of laryngeal carcinoma.

EGFR immunoexpression and peripheral blood cytokine secretion as potential biomarkers of tumor behavior in laryngeal squamous cell carcinoma.

BACKGROUND: Epidermal growth factor receptor (EGFR) and proinflammatory cytokines have been implicated in the dysregulated cell growth and increased aggressiveness of human cancers. The purpose of this study was to analyze EGFR immunoexpression in neoplastic tissues and IL-6 and TNFalpha secretion by peripheral blood mononuclear cells (PBMCs) and to investigate their relationships with certain clinicopathological characteristics. MATERIAL/METHODS: Tumor expression of membranous and cytoplasmic EGFR was measured in 45 cases of laryngeal squamous cell carcinoma by IHC staining. IL-6 and TNFalpha concentrations in 21-h PBMC cultures were measured by ELISA. Relationships between EGFR and cytokine secretion and clinicopathological characteristics such as pT status, pN status, and TFG classification, which include the parameters of the most invasive zones of neoplastic tissue and histological grade, were analyzed. RESULTS: The membranous EGFR index had a very strong association with pT stage, mode of invasion, and lymphocytic plasma infiltration of the tumor stroma. Relationships between the cytoplasmic EGFR index and nuclear polymorphism as well as TFG score for advanced carcinomas and histological grade in less invasive tumors were highlighted. The correlations of IL-6 and TNFalpha levels with TFG score, pT status, histological grade, and mode of tumor invasion were also significant. CONCLUSIONS: These findings confirmed the importance of EGFR immunoexpression rate as well as IL-6 and TNFalpha secretion by PBMCs as potential biomarkers for assessing the aggressive tumor phenotype in laryngeal carcinoma.

[Expression of transcription nuclear factor NFkappaB in laryngeal carcinoma tumor cells--correlation with IL-10 expression by blood mononuclear cells and clinicomorphological features]. / Ekspresja transkrypcyjnego czynnika jadrowego NFkappaB w komórkach raka krtani--korelacja z ekspresja IL-10 oraz cechami kliniczno-morfologicznymi guza.

PURPOSE: Transcription nuclear factor NFkappaB (p65-p50/RelA) is an activator of transcription process and regulates the apoptosis, cell cycle, proliferation, secretion of cytokines, angiogenic and growth factors by initiation gene transcription. NFkappaB can be activated by signaling pathway following IL-10 stimulation. The aim of this study was to estimate NFkappaB cytoplasmic and nuclear immunoexpression and to analyze the connection with clinicopathological features and IL-10 concentration produced by blood mononuclear cells in patients with laryngeal carcinoma. MATERIALS AND METHODS: Analysis of NFkappaB expression by immunmohistochemistry and IL-10 production by PBMCs and measured by Elisa in 45 patients with squamous cell carcinoma of the larynx were performed. Connections NFkappaB immunoexpression with clinicomorphological features (pT, pN, Anneroth, Batsakis i Lunas' classification) and IL-10 secretion were analyzed. RESULTS: Authors reported statistical correlation between NFkappaB cytoplasmic level and clinicomorphological parameters such as neoplasm advance according to Anneroth, Batsakis i Lunas' classification and depth of invasion as well as IL-10 secretion by PBMCs. CONCLUSION: Our studied indicated the important influence of NFkappaB activity on advance in laryngeal carcinoma as well as connection of anti-inflammatory and regulatory cytokine IL-10 produced by immunocompetent cells for course of neoplasm disease.

[Expression of activation antigens on the lymphocytes T in patients with carcinoma of the larynx and connection with tumor features]. / Ekspresja wybranych antygenów aktywacji na limfocytach T u chorych z rakiem krtani a cechy kliniczno-morfologiczne guza.

INTRODUCTION: Results of studies analyzing the role of immunocompetent cells in tumor environment and whole peripheral blood indicate their responsibility for aggressiveness of neoplasm, prognosis and therapeutic effect. Atcivation of lymhocytes T is connected with expression the markers (antigens) on their surface. The aim of this study was the analysis of activation antigens expression on lymphocytes T in patients with laryngeal carcinoma and the connection with clinicomorphological features. MATERIAL AND METHODS: Analysis of activation antigens expression CD69, CD71 and CD25, CD26, HLA/DR on lymphocytes T CD4+ i CD8+ in 33 patients with squamous cell carcinoma of the larynx was performed. Flow cytometry-based analysis of activation antigens in T cell cultures with and without PHA stimulation was used. The connection of these molecules and clinicomorphological features was examined (pT, pN, G, Anneroth, Batsakis and Lunas' classification). RESULTS: The significant correlation between chosen markers of activation and tumor features were noted: pT with HLA/DR/CD4, CD69CD8, CD71CD8, pN with CD26CD8, G with CD25CD8, CD71CD8, ABL score with CD25CD4. CONCLUSION: Our data indicated the connetion of immunocompetent cell activity and spread of neoplasm in patients with laryngeal carcinoma.

[Production of cytokines by peripheral blood mononuclear cells--correlation with clinicomorphological features in laryngeal carcinoma]. / Wydzielanie wybranych cytokin przez monocytarne komórki krwi--korelacja z cechami kliniczno-morfologicznymi w raku krtani.

INTRODUCTION: In studied analyzed role of the cytokines in pathology of neoplasms of various origin the importance of these proteins in regulation of immunocompetent cells function has been described. The aim of this study was to estimate of cho sen cytokines concentration produced by peripheral blood mononuclear cells and in whole blood in patients with laryngeal carcinoma and to analyze the connection of cytokines profile with clinicopathological features. MATERIALA AND METHODS: 55 patients with squamous cell carcinoma of the larynx treated at ENT Department Medical University of Lodz between 2003-2007 were analyzed. For estimation of cytokine secretion the cultures of isolated peripheral blood mononuclear cells (T lymphocytes) and the whole blood were established. Production of cytokines in supernatants was detected by Elisa. Connections with clinicomorphological features (pT, pN, Anneroth, Batsakis i Lunas' classification) were analyzed. RESULTS: Authors reported statistical correlation between chosen cytokines concentration and clinicomorphological parameters: pT and IL-2, IL-6, IL-8, TNFalpha produced by isolated cells and IL-2, IL-6, TNFa and IFNgamma in whole blood, pN and IL-8, IL-10, IFNgamma; ABL score and IL-6, TNFalpha, IFNgamma produced by isolated cells and IL-2, IL-6, IL-10, TNFalpha, IFNgamma in whole blood. CONCLUSION: Our studied indicated the important influence of proinflammatory and regulatory cytokines produced by immunocompetent cells for course of neoplasm disease, aggressiveness and advance in laryngeal carcinoma.

Fibroblast growth factor receptor 1 and 3 expression is associated with regulatory PI3K/AKT kinase activity, as well as invasion and prognosis, in human laryngeal cancer.

PURPOSE: Aberrant fibroblast growth factor receptor (FGFR) expression is thought to contribute to the development of many types of cancer. As yet, however, their impact on the course and prognosis of head and neck cancer remains to be determined. Here, we aimed to investigate the effects of expression of the FGFR family members FGFR1 and FGFR3, as well as their downstream PI3K/AKT signal-regulated kinases, on the aggressiveness and prognosis of laryngeal cancer. METHODS: In total 137 surgically removed squamous cell laryngeal cancer (SCLC) and 100 matched non-cancerous laryngeal mucosa (NCLM) samples were assessed for mRNA expression using quantitative real-time PCR. The corresponding proteins were analyzed by Western blotting. SLUG expression was assessed by immunohistochemistry. The expression data were subsequently related to tumor front grading (TFG), local/nodal recurrences, prognosis and overall survival. RESULTS: The FGFR1, FGFR3 and PI3K/AKT kinase mRNA and protein levels were found to be significantly higher in the SCLC than the NCLM samples (p < 0.05). A high FGFR1 mRNA/protein expression level was found to be associated with an increased invasion rate, according to TFG scale and SLUG level, a high local/nodal recurrence rate and a poor prognosis (p < 0.05). Similarly, we found that a high FGFR3 mRNA/protein expression level was associated with a shorter survival time (p < 0.05). In addition, we found that high PI3K/AKT kinase mRNA/protein levels were associated with a high TFG (p < 0.05). We also found that FGFR1/3 mRNA and FGFR1 protein levels were inversely associated with overall survival (log-rank test: FGFR1 mRNA p = 0.03, FGFR3 mRNA p = 0.04, FGFR1 protein p = 0.03). Subsequent multivariate analyses revealed that high FGFR3 mRNA expression may serve as an independent poor prognostic factor (HR 2.32, 95% CI 1.03-6.59; p = 0.04). We also found that the p-PI3K regulatory kinase protein level was significantly associated with survival in the cohort studied (HR 1.78, 95% CI 0.64-8.53; p = 0.03). CONCLUSIONS: From our data we conclude that FGFR1 and FGFR3, as well as its downstream regulatory PI3K/AKT kinases, may serve as potential biomarkers for the invasiveness and prognosis of laryngeal cancer. The expression of FGFR1/3-PI3K/AKT regulatory pathway members may be instrumental for the identification of patients at risk for an unfavorable clinical outcome.

[Correlation of membrane type I matrix metalloproteinase (MT1-MMP) expression with clinicomorphological features of tumor front in squamous cell carcinoma of the larynx]. / Korelacja ekspresji metaloproteinazy blonowej macierzy zewnatrzkomórkowej typu I (MT1-MMP) we froncie nacieku nowotworowego z cechami kliniczno--morfologicznymi w raku plalkonablolkowym krtani.

UNLABELLED: Matrix metalloproteases (MMPs) are proteolytic enzymes contribute to tumor expansion by degrading components of the extracellular matrix. MPPs play a key role in tumor invasion and metastases of various cancers and head and neck carcinoma as well. The aim of this study was to investigate MT1-MMP expression in squamous cell carcinoma of the larynx to relate these levels of expression to clinicohistological features of the tumor and lymph nodes e.g. TNM, nodal micrometastases, tumor front grading and 3- and 5-year survival. MATERIAL AND METHODS: The study included 22 patients with laryngeal cancer surgical treated in ENT Department Medical University of Lódz between 1998 and 1999. The expression of MT1-MMP was evaluated immunohistochemically using monoclonal antibodies anti-MT1-MMP (Mouse Anti-Human MT1-MMP, MAB3319 Chemicon). The polyclonal antibodies anti-CK for developing nodal micrometastases was used as well. RESULTS: Positive MT1-MMP expression in 68.2% cases was observed. Immunoexpression of MT1-MMP in advanced laryngeal carcinoma as indicator for 3-year survival was noted. In addition, levels of staining correlated with number of mitoses in tumor front and plasmocytolymphatic infiltration in its environment. CONCLUSIONS: The expression of MT1-MMP in tumor front appears to play an important role in determining prognosis in patients with carcinoma of the larynx.

[Clinico-histopathological characteristic of inverted papilloma of nasal cavity and paranasal sinuses cases treated in Department of Laryngological Oncology UM of Lodz between 2002-2006--review of latest literature]. / Kliniczno-histopatologiczna charakterystyka przypadków brodawczaka odwróconego jam nosa i zatok przynosowych leczonych w Klinice Laryngologii Onkologicznej UM w Lodzi w latach 2002-2006--przeglad najnowszego pismiennictwa.

Authors introduced diagnostic procedures, clinico-morphological features and treatment results of 16 cases of nasal cavity and paranasal sinuses inverted papillomas treated surgically on Department of Laryngological Oncology of Medical University of Lodz between 2002-2006. References review, definition, classification, clinico-hiostopathological criteria and treatment methods of Schneiderian papilloma were introduced in study.

[Morphological tumor front grading and matrix metalloproteinases type I expression as a prognostic parameter of the presence of lymph node micrometastases in laryngeal carcinoma]. / Morfologiczna ocena nacieku nowotworowego oraz ekspresji metaloproteinazy blonowej macierzy zewnatrzkomórkowej typu I jako prognostyczny wskaznik powstawania mikroprzerzutów w raku krtani.

INTRODUCTION: Occult foci of neoplasm cells in lymph nodes (referred to as micrometastases) in various squamous cell carcinomas may be discovered by immunohistochemistry by using anti-CKs (cytokeratine filaments) policlonal antibodies which reactive with epithelial cells. Matrix metalloproteinases (MMPs) are proteolytic family enzymes represent a group of endopeptidases which are capable to degrading components of the extracellular matrix and have been implicated as playing an important role in cancer invasion and metastases. The purpose of this study was to analyze the morphological parameters and to investigate MT1-MMP expression in laryngeal carcinoma to relate the expression to CKs in pN0 lymph nodes. MATERIALS AND METHODS: To presented the direct correlation between the morphological features of tumor front and the probability of micrometastases and prediction of prognosis we have analyzed 22 patients operated for squamous cell carcinoma of the larynx. The total score of TFG classification, tumor clinicomorphological features and grade of matrix metalloproteinase membrane type 1 staining in tumor front were analyzed to predict the presence of micrometastases and prognosis. Immunohistochemical methods with a panel of CKs antigens in lymph nodes and MT1-MMP expression in tumor tissue were performed. RESULTS: Our study showed that the total morphologic score TFG is very useful in the prediction of micrometastases in patients with laryngeal squamous cell carcinoma. The statistical analysis has revealed a significant correlation between the total TFG score and the depth of invasion and the presence of micrometastases. Positive MT1-MMP expression in 68.2% cases was observed. There was no significant relationship for immunoexpression of MT1-MMP in examined group of patients with advanced laryngeal carcinoma and positive poliCKs stain in lymph nodes. CONCLUSIONS: The results of study suggest that extended traditional pathologic evaluation by features from the TFG classification, especially the depth of invasion, could aid in diagnosis of micrometastases. The positive expression of poliCKs in the conventional pathological examination of pN0 lymph nodes appears to play an important role in determining prognosis in patients with carcinoma of the larynx.