Iwona Lewy-Trenda

The expression of SOCS1 and TLR4-NFkappaB pathway molecules in neoplastic cells as potential biomarker for the aggressive tumor phenotype in laryngeal carcinoma.

Suppressor of cytokine signaling 1 (SOCS1) is the key regulator of cytokine-mediated innate and adaptive immunity. One of the molecular mechanisms of SOCS1 is connected with inhibition of TLR4-NFkappaB pathway. The relationships among these molecules in laryngeal carcinoma are not exactly known. In this preliminary study we focused on their special activity and role in regulation of development and progression of laryngeal carcinoma. To investigate NFkappaB (p65 subunit) nuclear and cytoplasmic expression in 45 tumor samples of advanced laryngeal carcinoma IHC staining was performed. To determine the mRNA expression levels of TLR4, IRAK1, TRAF6 and SOCS1 in isolated neoplasm cells and non-cancerous adjacent mucosa epithelial cells RT-PCR was used. The invasiveness of laryngeal carcinomas was evaluated according to tumor front grading, TFG, which included tumor-related features (cytoplasmic differentiation, nuclear polymorphism, number of mitoses) and adjacent stroma-related characteristics of the peripheral edge of tumor infiltration (mode of infiltration, depth of invasion and plasmalymphocytic infiltration). The relationships between pT, pN status, the histological G grade, certain clinicopathological characteristics as well as postoperative observation time and the mRNA expression of the molecules mentioned earlier were investigated. Significant differences of TLR4-NFkappaB pathway molecules and SOCS1 mRNA expression in laryngeal tumor cells and normal adjacent mucosa cells as well as significant interconnections of TLR4, SOCS1 and NFkappaB(p65) in isolated tumor cells were obtained. This preliminary study demonstrated that the expression of SOCS1 and TLR4-NFkappaB pathway molecules had a strong association with the aggressiveness of laryngeal carcinoma. Positive relationships of TRAF6 in tumor margin cells with the histological grade and the mode of tumor invasion as well as the TFG total score were highlighted. Significant positive correlations were found between the TLR4 in tumor central cells and the TFG total score. Negative relationships of SOCS1 in tumor central cells with the histological grade were also noted. Significant positive correlations were found between the cytoplasmic NFkappaB(p65) and the mode of invasion as well as TFG total score. Our findings confirmed the importance of SOCS1 and TLR4-NFkappaB pathway molecules as potential biomarkers for assessment of the aggressive tumor phenotype 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.

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.

Kliniczno-histopatologiczna charakterystyka przypadków brodawczaka odwróconego jam nosa i zatok przynosowych leczonych w Klinice Laryngologii Onkologicznej UM w Łodzi w latach 2002–2006 – przegląd najnowszego piśmiennictwa

Summary 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, clinical and histopathological criteria and treatment methods of Schneiderian papilloma were introduced in study.

Ekspresja transkrypcyjnego czynnika jądrowego NFκB w komórkach raka krtani – korelacja z ekspresją IL-10 oraz cechami kliniczno-morfologicznymi guza☆☆☆◊◊◊

Summary Purpose Transcription nuclear factor NFκB (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. NFκB can be activated by signaling pathway following IL-10 stimulation. The aim of this study was to estimate NFκB 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 NFκB 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 NFκB immunoexpression with clinicomorphological features (pT, pN, Anneroth, Batsakis i Lunas’ classification) and IL-10 secretion were analyzed. Results Authors reported statistical correlation between NFκB 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 NFκB 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.

Secretion of inflammatory and regulatory cytokines and the association with depth and type of tumour invasion in patients with cancer of the larynx

prozapalnych, immunoregulatorowych, proonkogennych i prometastatycznych, wydzielane przez komórki układu odpornościowego oraz komórki nowotworowe, sprzyjają progresji choroby nowotworowej. Celem pracy była ocena wydzielania wybranych cytokin prozapalnych i regulatorowych, oznaczonych we krwi obwodowej u osób z rakiem krtani, w aspekcie ich zastosowania jako biomarkerów fenotypu większej agresywności guza. Materiał i metody: Analizą objęto grupę 106 osób, w tym 55 chorych ze zweryfikowanym rakiem płaskonabłonkowym krtani. W patomorfologicznej ocenie zaawansowania zmian (głębokość i sposób inwazji nowotworowej) uwzględniono kryteria klasyfikacji oceny zmian we froncie guza (tumour front grading – TFG). Stężenia cytokin, takich jak interleukiny (IL-6, IL-8 i IL-10), czynnik martwicy nowotworów (tumour necrosis factor – TNF), interferon γ (IFN-γ) we krwi obwodowej chorych, oceniano w dwóch punktach czasowych (21 godz. i 72 godz.) metodą immunoenzymatyczną ELISA. Zestawiono wyniki badań immunologicznych i kliniczno-morfologicznych. Wyniki: Analiza wyników przeprowadzonych badań wykazała, że w rakach krtani charakteryzujących się najwię k szą rozległością miejscową pT4 ob ser wowano znamiennie najmniejsze wy dzielanie IL-6 i IFN-γ oraz największe wydzielanie TNF. W guzach nowotworowych o największej głębokości inwazji, z naciekaniem warstwy lamina propria oraz chrząstki krtani stwierdzono znamiennie najmniejsze stężenia IL-6 i IFN-γ. W rakach krtani, które charakteryzowały się dobrze odgraniczonymi marginesami nacieku, stwierdzono znamiennie największe stężenia IL-6 i IL-10. Wnioski: Przeprowadzone badania wskazują na znaczenie aktywności cytokin w przebiegu choroby nowotworowej jako biomarkerów fenotypu zaawansowania i progresji zmian.

Ekspresja wybranych antygenów aktywacji na limfocytach T u chorych z rakiem krtani a cechy kliniczno-morfologiczne guza ☆ ☆☆ ◊ ◊◊ ★

Summary 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.

Wydzielanie wybranych cytokin przez monocytarne komórki krwi – korelacja z cechami kliniczno-morfologicznymi w raku krtani ☆ ☆☆

Summary 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 chosen 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. Materials 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, TNFα produced by isolated cells and IL-2, IL-6, TNFα and IFNγ in whole blood, pN and IL-8, IL-10, IFNγ; ABL score and IL-6, TNFα, IFNγ produced by isolated cells and IL-2, IL-6, IL-10, TNFα, IFNγ 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.

Morfologiczna ocena nacieku nowotworowego oraz ekspresji metaloproteinazy błonowej macierzy zewnątrzkomórkowej typu I jako prognostyczny wskaźnik powstawania mikroprzerzutów w raku krtani

Summary 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.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.

Morfologiczna ocena nacieku nowotworowego oraz ekspresji metaloproteinazy błonowej macierzy zewnątrzkomórkowej typu I jako prognostyczny wskaźnik powstawania mikroprzerzutów w raku krtaniMorphological tumor front grading and matrix metalloproteinases type I expression as a prognostic parameter of the presence of lymph node micrometastases in laryngeal carcinoma

Summary 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.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.