Marcin Fraczek

Epidermal differentiation complex (locus 1q21) gene expression in head and neck cancer and normal mucosa.

Epidermal differentiation complex (EDC) comprises a number of genes associated with human skin diseases including psoriasis, atopic dermatitis and hyperkeratosis. These genes have also been linked to numerous cancers, among them skin, gastric, colorectal, lung, ovarian and renal carcinomas. The involvement of EDC components encoding S100 proteins, small proline-rich proteins (SPRRs) and other genes in the tumorigenesis of head and neck squamous cell cancer (HNSCC) has been previously suggested. The aim of the study was to systematically analyze the expression of EDC components on the transcript level in HNSCC. Tissue specimens from 93 patients with HNC of oral cavity and 87 samples from adjacent or distant grossly normal oral mucosawere analyzed. 48 samples (24 tumor and 24 corresponding surrounding tissue) were hybridized to Affymetrix GeneChip Human 1.0 ST Arrays. For validation by quantitative real-time PCR (QPCR) the total RNA from all180 samples collected in the study was analyzed with Real-Time PCR system and fluorescent amplicon specific-probes. Additional set of samples from 14 patients with laryngeal carcinoma previously obtained by HG-U133 Plus 2.0 microarray was also included in the analyses. The expression of analyzed EDC genes was heterogeneous. Two transcripts (S100A1 and S100A4) were significantly down-regulated in oral cancer when compared to normal mucosa (0.69 and 0.36-fold change, respectively), showing an opposite pattern of expression to the remaining S100 genes. Significant up-regulation in tumors was found for S100A11, S100A7, LCE3D, S100A3 and S100A2 genes. The increased expression of S100A7 was subsequently validated by QPCR, confirming significant differences. The remaining EDC genes, including all encoding SPRR molecules, did not show any differences between oral cancer and normal mucosa. The observed differences were also assessed in the independent set of laryngeal cancer samples, confirming the role of S100A3 and LCE3D transcripts in HNC. In HNC of oral cavity only one family of EDC genes (S100 proteins) showed significant cancer-related differences. A number of other transcripts which showed altered expression in HNC require further validation.

A microarray study of gene expression profiles in nasal polyps

OBJECTIVE Nasal polyposis (NP) is a multifactorial disease manifesting in chronic inflammation of upper respiratory tract of unknown etiology. We studied mRNA gene expression profiles in NP compared with normal mucosa as well as pointed at genes characteristic of different expression in examined tissues. MATERIAL AND METHODS Fifty-three patients with NP (36 eosinophilic and 17 neutrophilic NP) were included into the study. Transcriptional activity of genes was analyzed using oligonucleotide microarray in 17 NP and 8 cases of normal nasal mucosa. A study of mRNA expression of selected genes was performed using QRT-PCR. RESULTS We identified 556 genes, which were differentially expressed between the studied and the control group. Among them 217 showed significantly higher expression, whereas 339 lower expression in NP than in controls. The microarray and QRT-PCR results were compatible for 7 of 8 evaluated genes. In NP strongly significant higher transcriptional activity of MMP10, NOS2A, ALOX15 and IL-8 genes was observed. In the control group, significantly higher expression of DMBT1, ALOX12 and LTF genes was detected. CONCLUSION The analysis of gene expression in inflammatory changed nasal polyp tissues may become a supplementary method in diagnostics and treatment. Molecular alterations may indicate changes during the clinical course of the disease.

Expression of c-myc oncoprotein in laryngeal squamous cell carcinoma.

Objective c-myc seems to play a pivotal role in normal growth and development as well in cellular transformation and carcinogenesis. Overexpression of the c-myc oncogene has been observed in many hematopoetic and solid tumors. The role of c-myc protein in squamous cell carcinoma of the head and neck in general and laryngeal squamous cell carcinomas (LSCCs) in particular is far from clear. The aim of this study was to evaluate the relations between the level of c-myc protein in LSCCs and the clinicopathological data of patients, DNA ploidy and the SG2M phase index (PI). Material and Methods The c-myc protein level was evaluated immunohistochemically in tumor specimens from 50 patients with LSCC. The DNA index and SG2M PI were determined by means of flow cytometry. Results We found c-myc protein in 34 (68%) tumors. Expression of c-myc protein was demonstrated to be frequent in non-metastatic cases (p=0.016). There was no association between c-myc protein level and age, primary tumor size, histological grading or type of cancer. In 13 (26%) cases we observed DNA aneuploid tumors. The mean value of the SG2M PI was 22.5%. Expression of c-myc protein was not related to SG2M PI or DNA ploidy. Conclusions We have shown that c-myc oncoprotein may be involved in the genesis of LSCC. Our findings suggest that the detectability of c-myc protein is associated with a lower metastatic potential. The c-myc oncogene is probably not as important in laryngeal cancers compared to other cancers. Further investigations must be performed to establish the value of predicting nodal metastases in LSCC.

Expression of collagenase-1 (MMP-1), collagenase-3 (MMP-13) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in laryngeal squamous cell carcinomas

Matrix metalloproteinases (MMPs) that have a proteolytic activity against the components of extracellular matrix (ECM) play an important role in the invasive and metastatic spread of tumors. The role of MMPs and tissue inhibitors of MMPs (TIMPs) in laryngeal squamous cell carcinoma (LSCC) has not been elucidated sufficiently. The aim of the present study was to evaluate the correlation between the expression of collagenase-1 (MMP-1), collagenase-3 (MMP-13) and TIMP-1, as well as the clinicopathological features of LSCCs. The expression of collagenases and TIMP-1 was examined immunohistochemically in 50 cases of surgically obtained specimens of primary LSCCs. Analyses indicated that LSCC cells as well as stromal cells expressed MMP-1, MMP-13 and TIMP-1 immunostaining. Overexpression of TIMP-1 occurred more frequently in non-metastasizing cases ( P =0.009). TIMP-1 and MMP-1 staining correlated significantly with the histologic type of LSCC. The keratinizing type of carcinomas exhibited higher TIMP-1 protein expression than the nonkeratinizing variety ( P =0.01). TIMP-1 staining was associated with the grade of differentiation, since it was found predominantly in well and moderately differentiated carcinomas ( P =0.04). The findings confirm that expression of analyzed MMPs and TIMP-1 is characteristic of LSCC and that these enzymes contribute to the progression of tumors. TIMP-1 upregulation might exhibit lower metastatic potential in LSCCs and is linked rather with an early stage of tumor progression. It seems also that TIMP-1 expression is dependent on the grade of differentiation.

Expression patterns of cyclin E, cyclin A and CDC25 phosphatases in laryngeal carcinogenesis

The aim of the research was to evaluate the expression levels of cyclin E/A and CDC25A/B during laryngeal carcinogenesis. The expression was demonstrated using immunohistochemistry in 46 cases of laryngeal cancer (LSCC), 23 epithelial dysplasias (ED) and 21 samples of normal mucosa (NM). The mean labeling indices (LI) for cyclin E in LSCC, ED and NM were 10.6, 4.9 and 0%; for cyclin A 27.2, 17.5 and 7%; for CDC25A 73.9, 53 and 32% and for CDC25B 36.5, 25.9 and 0%, respectively. A gradual increase in cyclin A and CDC25A expression levels from mild through moderate and severe dysplasia to in situ carcinoma were noted. Cyclin A LI significantly increased also from NM through ED to LSCC. Cyclin A and CDC25A LI significantly increased from NM to ED. Overexpression of cyclin A and CDC25A was significantly associated with proliferation among ED. Linear interdependency was detected in ED between the expression of CDC25A and cyclin A. Cyclin E and CDC25B overexpression occurs as a late event in neoplastic transformation. The progressive expression of proteins supports the multistep model of laryngealcarcinogenesis. The results indicate a possible role of cyclin A as a marker reflecting cell proliferation. The enhanced immunoexpression of cyclin A and CDC25A suggests the potential for malignant formation in preneoplastic lesions.

Clinicopathologic significance and prognostic role of cyclin E and cyclin A expression in laryngeal epithelial lesions.

Conclusions. The determination of cyclin A expression might be helpful in the identification of laryngeal squamous cell cancer (LSCC) patients with increased risk of metastases. The results suggest that cyclin A may be a more informative marker for cell proliferation than Ki-67. Abnormalities of cyclin E and cyclin A may play an important role in LSCC development and progression; however, the expression of cyclin E does not seem to have prognostic significance. Objective. The aim of the study was to elucidate a possible association between cyclin E and cyclin A expression and clinicopathologic factors and their potential role as prognostic markers for patients with laryngeal epithelial lesions. Materials and methods. Expression of cyclins E and A, and Ki-67 was examined immunohistochemically in a formalin-fixed, paraffin-embedded series of 46 LSCC; 23 epithelial dysplasias (ED); and 21 normal mucosae (NM). Results. The mean labeling indices (LIs) for cyclin E in LSCC, ED, and NM were 10.6%, 4.9%, and 0%, and for cyclin A 27.2%, 17.5%, and 7%, respectively. In LSCC, a statistically significant correlation was found between enhanced cyclin A expression and a higher incidence of locoregional lymph node metastasis (p<0.01). The enhanced expression of cyclin A was linked with cell proliferation in LSCC, ED, and NM. No association was observed between cyclin E and A and other clinicopathologic parameters or applied treatments. The prognostic significance of cyclin E, cyclin A, and Ki-67 in determining overall survival time showed no statistically significant differences.

Fuzzy rule-based classification system for computer-aided diagnosis in contact endoscopy imaging

In this paper the image analysis system designed for contact endoscopy (CE) image interpretation is presented. The morphometric analysis of segmented nuclei is followed by the fuzzy rule-based classification. Rules generation is realized basis of fuzzy clustering approach. The classification results provide quantitative and qualitative interpretation and would be very helpful in the computer-aided cancer diagnosis of the larynx.