Zellen der Tumorfront

Background

With a frequency of about 90?%, head and neck squamous cell carcinomas (HNSCCs) are the most common malignancies of the upper aerodigestive tract. The cancer stem cell (CSC) hypothesis postulates that CSCs are the dangerous part of the tumor and are relevant to metastasis, invasiveness and resistance to chemotherapy.

Methods

Tissue samples taken from HNSCCs and normal mucosa were tested for the expression of several established CSC markers. The expression and activity of the matrix metalloproteinase MMP-9 was also investigated.

Results

Cells of the invasive tumor front expressed the basal stem cell markers CD44, ALDH1 and CK14. However, in contrast to the noninvasive basal cell layer of normal mucosa, HNSCC samples were also positive for active MMP-9, which lends the tumor its gelatinolytic activity.

Conclusion

These observations suggest a model in which cells of the invasive front are derived from the basal cell layer of normal mucosa and harbour the CSCs. Future studies should thus focus on the cells of the invasive front in particular, since the activity of these cells may form the basis for tumor recurrence and therapy resistance.     

Characterization of FaDu-R,a radioresistant head and neck cancer cell line,and cancer stem cells

Objectives

The aim of this study was to evaluate the impact of CSC on insensitivity to radiotherapy in HNSCC.

Primäre nasale NK/T-Zell-Lymphome

Background

Nasal NK/T-cell lymphomas are rare malignancies in Europe or North America. Histological diagnosis is difficult, because tumors imbedded in large necrotic areas and neoplastic infiltrates may be admixed with small lymphocytes, plasma cells, eosinophils, and histiocytes, and thus the process could be misdiagnosed as chronic inflammation. Progression of the disease leads to septal perforation and may also result in destruction of the hard palate, and if left untreated it ends fatally. This introduced the term “lethal midline granuloma”, a term which should not be used any more.

Material and methods

Clinical features, pathohistology, and current classification of primary nasal NK/T-cell lymphomas are described against the background of the recent literature and a case report.

Results

Immunophenotyping is essential for the diagnosis. Tumor cells are uniformly infected by Epstein-Barr virus, which could be verified by EBER in situ hybridization. Immunohistochemically, tumor cells are positive for CD56, cytoplasmic CD3ε, and CD2 and they express cytotoxic molecules like granzyme B, TIA-1, GMP17, and perforin. Therapeutic options are radio- or radiochemotherapy. On average, 2- and 5-year survival rates of 50% are obtained in stages I and II. The prognosis of advanced tumor stages is very poor.

Conclusions

Immunohistochemical and molecular genetic early diagnosis is of crucial prognostic relevance.     

Selective Delivery of a Therapeutic Gene for Treatment of Head and Neck Squamous Cell Carcinoma Using Human Neural Stem Cells

Objectives

Based on studies of the extensive tropism of neural stem cells (NSCs) toward malignant brain tumor, we hypothesized that NSCs could also target head and neck squamous cell carcinoma (HNSCC) and could be used as a cellular therapeutic delivery system.

Methods

To apply this strategy to the treatment of HNSCC, we used a human NSC line expressing cytosine deaminase (HB1.F3-CD), an enzyme that converts 5-fluorocytosine (5-FC) into 5-fluorouracil (5-FU), an anticancer agent. HB1. F3-CD in combination with 5-FC were cocultured with the HNSCC (SNU-1041) to examine the cytotoxicity on target tumor cells in vitro. For in vivo studies, an HNSCC mouse model was created by subcutaneous implantation of human HNSCC cells into athymic nude mice. HB1.F3-CD cells were injected into mice using tumoral, peritumoral, or intravenous injections, followed by systemic 5-FC administration.

Results

In vitro, the HB1.F3-CD cells significantly inhibited the growth of an HNSCC cell line in the presence of the 5-FC. Independent of the method of injection, the HB1.F3-CD cells migrated to the HNSCC tumor, causing a significant reduction in tumor volume. In comparison to 5-FU administration, HB1.F3-CD cell injection followed by 5-FC administration reduced systemic toxicity, but achieved the same level of therapeutic efficacy.

Conclusion

Transplantation of human NSCs that express the suicide enzyme cytosine deaminase combined with systemic administration of the prodrug 5-FC may be an effective regimen for the treatment of HNSCC.     

Charakterisierung von Stammzellen des neonatalen auditorischen Sinnesepithels

Background

In contrast to regenerating hair cell-bearing organs of nonmammalian vertebrates the adult mammalian organ of Corti appears to have lost its ability to maintain stem cells. The result is a lack of regenerative ability and irreversible hearing loss following auditory hair cell death. Unexpectedly, the neonatal auditory sensory epithelium has recently been shown to harbor cells with stem cell features. The origin of these cells within the cochlea’s sensory epithelium is unknown.

Material And Methods

We applied a modified neurosphere assay to identify stem cells within distinct subregions of the neonatal mouse auditory sensory epithelium. Sphere cells were characterized by multiple markers and morphologic techniques.

Results

Our data reveal that both the greater and the lesser epithelial ridge contribute to the sphere-forming stem cell population derived from the auditory sensory epithelium. These self-renewing sphere cells express a variety of markers for neural and otic progenitor cells and mature inner ear cell types.

Conclusion

Stem cells can be isolated from specific regions of the auditory sensory epithelium. The distinct features of these cells imply a potential application in the development of a cell replacement therapy to regenerate the damaged sensory epithelium.     

Neurogene Stammzelltransplantation in die Kochlea

Background

Stem cell therapy is especially interesting for inner ear related diseases, since the hair cells are very sensitive and do not regenerate. Hair cell loss is therefore irreversible and is accompanied by hearing loss. In the last few years, different research groups have transplanted stem cells into the inner ear with promising results. In the presented study, our aim was to gain insight into how neuronal stem cells behave when they are transplanted, both in vitro and in vivo, into a damaged inner ear.

Methods

Neuronal stem cells from E9.5 day old mouse embryos were collected and infected with an adenoviral vector encoding green fluorescent protein (GFP). GFP+ cells were then transplanted into a damaged organ of Corti in vitro or into a damaged mouse inner ear in vivo.

Results

We were able to detect GFP+ cells close to the organ of Corti in vitro and in the organ of Corti in vivo. The GFP+ cells do not seem to be randomly distributed in either the in vitro or in vivo situation. Most interestingly, GFP+ cells could be detected close to places where hair cells had been lost in vivo.

Conclusion

Neuronal stem cells are interesting candidates to replace lost hair cells. However, a great deal of research is still needed before they can enter clinical trials.     

Pr?valenz auff?lliger Schleimhautbefunde bei Patienten in HNO-Praxen

Objective

To date, no secondary prevention program is in place for patients carrying an increased risk for developing head and neck cancer (HNSCC). In terms of successful, long-term curative therapy and increased quality of life, it would be useful to detect such diseases at an early stage.

Patients and Methods

A total of 370?patients with at least one risk factor such as ??smoking??, ??alcohol??, or ??reflux disease?? and without any symptoms were examined during a 1-year period using standard HNO methods (e.?g. endoscopy) for suspicious alterations of the mucosa of the upper aerodigestive tract.

Results

In 13 (3.5%) of all 370 cases a biopsy was taken for further diagnosis. Squamous cell carcinoma was found in eight cases, while one further patient was suffering from non-Hodgkin lymphoma.

Conclusions

It is simple and safe to examine patients at risk of developing HNSCC by standard HNO methods. The rate of detected carcinomas is much higher than in former investigations, likely because our survey focused only on patients with specific risk factors.     

Gezüchtete Knorpeltransplantate

Background

The main technique used in tissue engineering for the generation of autologous cartilage grafts is the production of autologous transplant material from living cells or tissues and/or cell matrices. Incompletely absorbed residual fibrous matter, unforeseeable interactions between cells and biological materials and uneven cell distribution of cells in the cell carriers still present unsolved problems.

Methods

For these reasons a three-dimensional aggregate culture system was developed in which cells can generate cartilaginous tissue without the use of biomaterials. Chondrocytes and adult mesenchymal stem cells were used for this purpose and generate cartilaginous tissue with various phenotypes both in the aggregate culture system and in the athymic nude mouse model. The newly generated cartilage was subjected to histomorphological, immunochemical and biochemical investigation.

Results

After 3 weeks of in vitro aggregate culture the chondrocytes of all subclasses formed cartilaginous tissue. After 6 weeks’ in vivo maturation in the athymic nude mouse model the new cartilage was found to differ in typical phenotype depending on the native cartilage used.

Conclusions

Cartilage cells of various subclasses and adult menchymal stem cells generate cartilaginous tissue corresponding to their own phenotypes in a 3D aggregate culture system. This culture system is a promising method of producing cartilage grafts for use in reconstructive head and neck surgery.     

Krebshemmende Wirkung des ?therischen Salbei-?ls auf eine Plattenepithelzellkarzinom-Zelllinie der Mundh?hle (UMSCC1)

Background

Every year there are several hundred thousand new cases of oral cancer worldwide. Clinical oncology is still challenged by toxicity and side effects of multimodal therapy strategies in which is associated with poor prognosis for patients. There is an urgent necessity to develop novel therapy strategies. As the majority of anticancer drugs are of natural origin, natural products represent a valuable source for the identification and development of novel treatment options for cancer. The aim of this investigation was to study the cytotoxicity of Salvia officinalis L. (sage) essential oil.

Methods

Salvia officinalis essential oil was gained by aqueous extraction from plant material and subsequently analyzed by gas chromatography. The cytotoxicity of the essential oil on the squamous human cell carcinoma cell line of the oral cavity (UMSCC1) was assessed with the XTT assay. These experiments revealed the half maximal inhibitory concentration (IC₅₀) of the essential oil. It was used in the microarray-based analysis of gene expression of UMSSC1 cells. The results were submitted to a signaling pathway analysis.

Results

The main constituents of Salvia officinalis essential oil include the monoterpenes thujone, ??-pinene, and 1,8-cineol. Low concentrations of the essential oil increased vitality of the UMSCC1 cells. Beyond the concentration of the IC₅₀ of 135???g/ml, sage essential oil reduced UMSSC1 cells viability to a minimum. In the microarray gene expression analysis, genes involved in cancer, cellular growth and proliferation, cell death, cell morphology, cell cycle, gene expression, and DNA repair were the most prominent. The three most significantly regulated pathways by sage were aryl hydrocarbon receptor signaling, cell cycle (G1/S checkpoint) regulation, and p53 signaling.

Conclusion

To the best of our knowledge, this study suggests for the first time the ability of Salvia officinalis essential oil to inhibit human HNSCC cell growth. The therapeutic potential of sage essential oil might exceed that of its common use in otorhinolaryngology.     

Solitärer fibröser Tumor der Nasenhaupt- und Nasennebenhöhlen

Objective

The solitary fibrous tumor is an uncommon, benign lesion with a mesenchymal origin which arises most commonly from the pleura and, in extremely rare cases, from the mucosa of the nose and paranasal sinuses.

Patient and methods

We describe a case of solitary fibrous tumor in the nasal cavity with an extension into both ethmoid sinuses and destruction of the nasal septum in a 64-year-old woman presenting with nasal obstruction and frontal headache. In the histopathologic examination and by immunohistochemistry, the tumor was composed of spindle cells and nodular collagen, and was confirmed as a solitary fibrous tumor. It was completely removed using an endonasal approach with the patient being free of any complaints and without evidence of disease 2 years after surgery.

Conclusions

Solitary fibrous tumor should be included in diagnostic considerations in patients with sinonasal masses, especially in cases with the appearance of spindle cells. Diagnostic procedures, clinicopathological features, therapeutic options and prognosis of the such tumors are discussed.     

Ectonucleotidase CD39 expression in regional metastases in head and neck cancer

Introduction: CD39 is the rate-limiting enzyme in the generation of immunosuppressive adenosine and its expression and activity are significant in tumor progression. Squamous cell carcinoma of the head and neck (HNSCC) shows an overall poor prognosis due to high local recurrence rates and early metastatic spread.

Material and methods: Primary tumor specimens and lymph node specimens harvested during neck dissection of 65 patients with a diagnosis of HNSCC were subjected to immunohistochemical and H-score analysis of CD39 expression. Demographics, histopathology and subsequent outcome were analyzed.

Results: The primary cancer was squamous cell carcinoma in all patients (male/female 55:10). H-score for CD39 expression in the primary lesion and metastatic lymph nodes was significantly higher in advanced compared to early stages with no significant differences among different tumor locations. High intratumoral and intrametastatic CD39 expression was associated with an inferior patients’ overall survival at a mean follow-up of 83.4 months (6–204 months).

Conclusion: CD39 expression in HNSCC correlated positively with tumor stage and appears to predict poor prognosis. Therefore, CD39 expression in primary lesions and metastatic lymph nodes seems to identify patients at high risk in HNSCC of all tumor sites. Immunotherapeutic approaches targeting CD39 might be promising for this patient population.     

Selektive Halslymphknotenausräumung beim primär klinisch und histopathologisch metastasenfreien Plattenepithelkarzinom der Mundhöhle

Background

The aim of this retrospective study was to evaluate prognostic factors for the development of secondary local lymph node metastases in patients with oral squamous cell carcinoma who had undergone selective neck dissection for primary node-negative disease.

Patients and Methods

The study included 331 patients with primary squamous cell carcinoma of the oral cavity who underwent 431 selective neck dissections between January 1986 and December 2002 in Germany at the Hannover Medical School’s Department of Oral and Maxillofacial Surgery. Several potential prognostic factors were evaluated for their influence on the development of secondary metastases following primary neck dissection.

Results

No statistically significant relationship to the appearance of secondary local metastasis following selective neck dissection was detected concerning: patient age or sex, histopathologic tumor stage, primary tumor grade, or adjuvant therapies such as pre- or postoperative radiotherapy and radiochemotherapy. The only study factor with a statistically significant influence was the extent of lymphadenectomy, in which particularly the region of the carotid bifurcation played a decisive role.

Conclusion

Significantly fewer secondary metastases occurred following neck dissections that included the carotid trigone. In light of these results, we recommend that neck dissection for primary oral squamous cell cancer always include the region of the carotid bifurcation, regardless of the above mentioned associated patient and tumor factors.     

Head and neck squamous carcinomas with exophytic and endophytic type of growth have the same prognosis after surgery and adjuvant radiotherapy

Head and neck squamous cell carcinomas (HNSCC) are characterized by exophytic or endophytic growth. We hypothesized that the growth pattern predicts outcome and associates with distinct clinical and immunological profiles. Tumors obtained from 60 HNSCC patients treated with surgery and adjuvant radiotherapy were identified as exophytic or endophytic. Recurrence-free survival (RFS) at 42 months was determined. In a subsets of 30 patients (22 exophytic and 8 endophytic) tumor stroma and parenchyma were evaluated for infiltrating CD4⁺ and CD8⁺ T, dendritic, myeloid and FOXP3⁺ regulatory T cells (Treg) and expression of immunosuppressive cytokines by immunohistochemistry. The localization and frequency of positive cells were determined microscopically and analyzed by hierarchical clustering to distinguish exophytic versus endophytic tumors. 34/60 patients had exophytic and 26/60 endophytic tumors. No differences in clinicopathologic data, disease progression or RFS were seen between the two cohorts. Infiltrates of CD3⁺CD8⁺ T cells were larger in endophytic than exophytic tumors, while FOXP3⁺ Treg, TGF-β⁺, IL-10⁺, Arg-1⁺, CD11b⁺ cells were equally prominent in both. FOXP3⁺ Treg accumulated in endophytic tumor nests, while the exophytic tumor stroma was enriched in IL-10⁺ cells (both at p < 0.05). Hierarchical clustering based on immunophenotyping failed to identify different clusters in these two tumor types. However, CD68⁺ macrophages and FOXP3⁺ Treg showed a distinct distribution. The HNSCC growth pattern did not predict RFS. Although higher numbers and differences in localization of immunosuppressive cells in endophytic versus exophytic tumors were observed, no significant relationship was established between the growth pattern and the immune profile of infiltrating lymphocytes.     

Wertigkeit der konventionellen oralen Bürstenbiopsie

Background

This study evaluated the performance of oral brush biopsies using standard morphological analysis and haematoxylin and eosin (HE) staining for detecting oral squamous cell carcinomas and their respective precursor lesions

Patients and methods

Brush biopsies were obtained in 169 consecutive patients who underwent routine biopsies and histological examination for clinically suspicious oral lesions. Air-dried smears were processed by acetone fixation and HE staining. Cytological assessment used well-established criteria of atypia to classify the specimen as either “tumor negative” (no signs of atypia, no malignant cells) or “tumor positive” (malignant cells, any sign of atypia or doubtful cells).

Results

Despite a sufficient number of cells, a definite cytological diagnosis could not be established in six cases. According to the criteria specified above, these specimens were classified as “tumor positive.” The cytological analysis identified 49 out of 62 oral malignancies (sensitivity 79%). Seven out of 107 benign lesions were classified as false positive (specificity 93%). The positive and negative predictive values were each 88%.

Conclusion

Oral brush biopsies will identify only about 80% of oral malignancies when the smears are processed by routine HE stains and are analysed via standard morphological criteria. Thus, this technique should not be used for diagnostic proof or to exclude malignant cells in a lesion suspicious for cancer. However, oral brush biopsy provides a versatile back-up strategy to uncover the true nature of the disease if a lesion is clinically considered benign by mistake.     

Serumkonzentrationen der löslichen Adhäsionsmoleküle sICAM-1 und sVCAM-1 bei Patienten mit malignen HNO-Tumoren

Background. Adhesion molecules are implicated in various stages of tumor progression and metastasis. Soluble forms of intercellular adhesion molecule-1 (sICAM-1) and vascular cell adhesion molecule-1 (sVCAM-1) were recently described. The aim of this study was to determine a differences in concentration of these molecules in patients with head and neck squamous cell carcinoma (HNSCC) compared to healthy controls. Patients and methods. We investigated the circulating levels of sICAM-1 and sVCAM-1 in sera from patients with various head and neck squamous cell carcinomas (n=52) as well as from normal, healthy controls (n=35). Serum concentrations were determined as serum immunoreactivity by using a quantitative sandwich enzyme immunoassay technique. For statistical analysis, the Student's t-test was performed. Results. The majority of the patients with HNSCC were found to have high concentrations of sICAM-1 and sVCAM-1. The mean level of sICAM-1 in patients was 313 ng/ml and in the control group 237 ng/ml (p=0.0005). The mean level of sVCAM-1 in patients was 624 ng/ml and in the control group 435 ng/ml (p=0.009). The concentrations of sICAM-1 and sVCAM-1 were significantly higher in HNSCC patients. Conclusions. These results show the difference in expression of these adhesion molecules in patients with HNSCC compared to healthy controls. Endothelial adhesion molecule determination may find clinical applications in the follow-up of cancer therapy. For this reason, studies incorporating the longitudinal follow-up of patients are required.     

Trägt Nikotin zur Krebsentstehung im oberen Aerodigestivtrakt bei?

Background

It is accepted that nicotine in tobacco smoke causes addiction via nicotinic acetylcholine receptors in the central nervous system. For a long time, the tumorigenic potential of smoking was attributed to compounds other than nicotine. However, more recently data have accumulated which suggest that nicotine may add to the cancer risk by stimulating cellular growth via non-neuronal acetylcholine receptors, by suppressing apoptosis, and by inducing angiogenesis not only in atheromatous plaques but also in tumors. In the present study the possible direct genotoxic effects of nicotine on DNA were investigated in human target cells of carcinogenesis in the upper aerodigestive tract.

Patients and methods

Human nasal mucosa, lymphatic tissue of the palatine tonsils, supraglottic epithelium of the larynx, and peripheral lymphocytes were exposed to rising concentrations of nicotine. DNA damage was investigated by alkaline single-cell microgel electrophoresis (Comet) assay. Cytotoxicity was assessed by trypan blue exclusion.

Results

Nicotine induced dose-dependent DNA damage in all cell types at low cytotoxic concentrations that allowed viabilities well above 80%. The lowest nicotine concentrations eliciting a significant increase in DNA migration were 1 mM for tonsillar cells and 0.25 mM for all other cell types.

Conclusion

Nicotine induces genotoxic effects in human target cells of carcinogenesis in the upper aerodigestive tract at relevant concentrations. Thus, nicotine may contribute directly to tumor initiation resulting from smoking.     

Qualitätssicherung nach rekonstruktiver Mittelohrchirurgie

Background

The ongoing improvement of imaging quality nowadays focuses increasingly more on immediate quality evaluation of surgical measures.

Methods

Rotational tomography (RT) has been experimentally proven to be an alternative imaging technique to computed tomography (CT) in terms of quality and postoperative localization of middle ear implants in temporal bones. Based on these results imaging parameters were determined in the form of a surgical check list in order to systematically check individual but important surgical reconstructive steps.

Results

The positive benefit of RT concerning immediate and significant postoperative quality control was confirmed by a patient study. The surgical check list was successfully implemented under clinical framework requirements. Among these standardized parameters the determined angulation of inserted middle ear implants was given outstanding significance due to the predictive value concerning functional outcome.

Conclusion

The RT procedure has been proven to be a reliable imaging tool for postoperative quality control following reconstructive middle ear surgery.     

Expression of VEGF, HGF, IL-6, IL-8, MMP-9, Telomerase in Peripheral Blood of Patients with Head and Neck Squamous Cell Carcinoma

Objectives

This study investigated the telomerase expression in peripheral blood mononuclear cells (PBMCs) and the relationship between the serum level of several soluble factors such as vascular endothelial growth factor (VEGF), hepatocyte growth factor, interleukin (IL)-6, IL-8, and matrix metallopeptidase-9 and the clinicopathological features of patients with head and neck squamous cell carcinoma (HNSCC).

Methods

Peripheral blood samples were collected from 50 HNSCC patients and 15 normal controls. The telomerase activity in the PBMCs was measured by Telomere Repeat Amplification Protocols. The serum levels of the soluble factors were analyzed by enzyme-linked immunosorbent assay.

Results

The expression of telomerase in the PBMCs of HNSCC patients was significantly correlated with the N and American Joint Committee on Cancer (AJCC) stages. The serum VEGF level was significantly higher in the patients with an advanced T stage, N stage and AJCC stage. Serum VEGF was significantly related with the expression of telomerase in the PBMCs. The telomerase expression and the VEGF expression were shown to be independent factors associated with poor survival.

Conclusion

The telomerase expression in the PBMCs and the serum VEGF level of HNSCC patients were significantly correlated with the N stage, the AJCC stage and the prognosis.