Immunohistochemical detection of PIEZO1 and PIEZO2 in human digital Meissner´s corpuscles.

BACKGROUND: The cutaneous end organ complexes or cutaneous sensory corpuscles are specialized sensory organs associated to low-threshold mechanoreceptors. Mechano-gated proteins forming a part of ion channels have been detected in both the axon and terminal glial cells of Meissner corpuscles, a specific cutaneous end organ complex in the human glabrous skin. The main candidates to mechanotransduction in Meissner corpuscles are members of the Piezo family of cationic ion channels. PIEZO2 has been detected in the axon of these sensory structures whereas no data exists about the occurrence and cell localization of PIEZO1. METHODS: Skin samples (n = 18) from the palmar aspect of the distal phalanx of the first and second fingers were analysed (8 female and 10 males; age range 26 to 61 26-61 years). Double immunofluorescence for PIEZO1 and PIEZO2 together with axonal or terminal glial cell markers was captured by laser confocal microscopy, and the percentage of PIEZOs positive Meissner corpuscles was evaluated. RESULTS: MCs from human fingers showed variable morphology and degree of lobulation. Regarding the basic immunohistochemical profile, in all cases the axons were immunoreactive for neurofilament proteins, neuron specific enolase and synaptophysin, while the lamellar cells displayed strong S100P immunoreactivity. PIEZO1 was detected co-localizing with axonal markers, but never with terminal glial cell markers, in the 56% of Meissner corpuscles; weak but specific immunofluorescence was additionally detected in the epidermis, especially in basal keratinocytes. Similarly, PIEZO2 immunoreactivity was found restricted to the axon in the 85% of Meissner corpuscles. PIEZO2 positive Merkel cells were also regularly found. CONCLUSIONS: PIEZO1 and PIEZO2 are expressed exclusively in the axon of a subpopulation of human digital Meissner corpuscles, thus suggesting that not only PIEZO2, but also PIEZO1 may be involved in the mechanotransduction from low-threshold mechanoreceptors.

Ag- and Sr-enriched nanofibrous titanium phosphate phases as potential antimicrobial cement and coating for a biomedical alloy.

Biomaterials and their surfaces regulate the biological response and ultimately the quality of healing at a possible site of implantation. The physical, chemical and topographical properties of implants' surfaces play a decisive role in the biological integration process for their immediate loading and long-term success. Since at this level of biological interaction nano-dimensionality is basically entailed, bio-functional nanostructured composites either as filling/cement or coating to metallic implants are required. This study shows the possibility of synthesizing two phases of nanostructured titanium phosphate (π and ρ polymorphs) and enriching them with silver nanoparticles and strontium. More importantly, Ag-Sr-enriched nanostructured π­titanium phosphate is induced to grow on a commercially available titanium alloy (Ti-6Al-4V), widely used in orthopedic and dental implants, under highly controlled conditions. Structural and microscopic studies, using XRD, HRTEM and SEM altogether confirm the resultant phases and their enrichment with strontium and silver nanoparticles with an average particle size around 6 nm. Using confocal laser scanning microscopy, the surface roughness was measured and is found to lay at the interface between the nanosized and microsized topologies. Ion release assessments showed that the presence of strontium controlled the release rate of silver ions and this could be beneficial in terms of decreasing the accompanied cytotoxicity that is usually encountered at high concentrations of silver release. Antimicrobial and cell proliferation assays have proved that enriching titanium phosphate with strontium and silver nanoparticles has improved their antimicrobial properties, while the cytotoxicity could be controlled.

Screening of Bifidobacteria and Lactobacilli Able to Antagonize the Cytotoxic Effect of Clostridium difficile upon Intestinal Epithelial HT29 Monolayer.

Clostridium difficile is an opportunistic pathogen inhabiting the human gut, often being the aetiological agent of infections after a microbiota dysbiosis following, for example, an antibiotic treatment. C. difficile infections (CDI) constitute a growing health problem with increasing rates of morbidity and mortality at groups of risk, such as elderly and hospitalized patients, but also in populations traditionally considered low-risk. This could be related to the occurrence of virulent strains which, among other factors, have high-level of resistance to fluoroquinolones, more efficient sporulation and markedly high toxin production. Several novel intervention strategies against CDI are currently under study, such as the use of probiotics to counteract the growth and/or toxigenic activity of C. difficile. In this work, we have analyzed the capability of twenty Bifidobacterium and Lactobacillus strains, from human intestinal origin, to counteract the toxic effect of C. difficile LMG21717 upon the human intestinal epithelial cell line HT29. For this purpose, we incubated the bacteria together with toxigenic supernatants obtained from C. difficile. After this co-incubation new supernatants were collected in order to quantify the remnant A and B toxins, as well as to determine their residual toxic effect upon HT29 monolayers. To this end, the real time cell analyser (RTCA) model, recently developed in our group to monitor C. difficile toxic effect, was used. Results obtained showed that strains of Bifidobacterium longum and B. breve were able to reduce the toxic effect of the pathogen upon HT29, the RTCA normalized cell-index values being inversely correlated with the amount of remnant toxin in the supernatant. The strain B. longum IPLA20022 showed the highest ability to counteract the cytotoxic effect of C. difficile acting directly against the toxin, also having the highest capability for removing the toxins from the clostridial toxigenic supernatant. Image analysis showed that this strain prevents HT29 cell rounding; this was achieved by preserving the F-actin microstructure and tight-junctions between adjacent cells, thus keeping the typical epithelium-like morphology. Besides, preliminary evidence showed that the viability of B. longum IPLA20022 is needed to exert the protective effect and that secreted factors seems to have anti-toxin activity.

Establishment and genetic characterization of six unique tumor cell lines as preclinical models for sinonasal squamous cell carcinoma.

Sinonasal squamous cell carcinomas (SCC) are rare tumors, etiologically related to occupational exposure to wood and leather dust. In spite of surgical and radiotherapeutic advances, the 5 year survival is still 30-50%. Therefore, alternative treatment options are needed. We report the establishment and characterization of six unique human sinonasal SCC cell lines, named SCCNC1, 2, 4, 5, 6 and 7. In vitro growth and invasion characteristics were evaluated and genetic profiles were compared to those of the original primary tumors. The population doubling times ranged from 21 to 34 hours. Cell lines SCCNC2 and 7 were highly invasive in matrigel. Five cell lines carried a high number of copy number alterations, including amplifications and homozygous deletions, while one showed only three abnormalities. Sequence analysis revealed three cell lines with TP53 mutation and none with KRAS or BRAF. Overexpression of p53 was observed in five, and of EGFR in four cell lines. None of the cell lines showed strong immunopositivity of p16 or presence of human papilloma virus. In conclusion, we have created six new cell lines that are clinically and genetically representative of sinonasal SCC and that will be a useful tool for the preclinical testing of new therapeutic agents.

From laryngeal epithelial precursor lesions to squamous carcinoma of the larynx: the role of cell cycle proteins and ß-catenin.

Novel markers to accurately predict the risk of malignant transformation in laryngeal epithelial precursor lesions (EPL) are needed. We tried to identify some molecular alterations occurring in laryngeal tumorigenesis. In this study, 60 paraffin-embedded EPL and 17 metachronous invasive carcinomas were immunostained for markers associated with proliferation (Ki67), cell cycle control (p53, p21, p16, p27, cyclin D1), and cell adhesion and invasion (laminin and ß-catenin). Aberrant expression of p16 and p53 and positivity at cytoplasm for ß-catenin and cyclin D1 were detected significantly in EPL with progression to invasive laryngeal carcinoma. All cases with basal and suprabasal reactivity of p53 showed ß-catenin overexpression. We found that ß-catenin protein expression increased significantly with the grade of dysplasia. This is one of the studies with the largest number of laryngeal EPL and invasive carcinoma studied sequentially. Our data confirm the role of some cell cycle regulatory proteins in the development of laryngeal carcinoma. Cytoplasmic retention of ß-catenin in EPL seems to be related with more aggressive biological behavior. Combined increased p53 and cytoplasmic ß-catenin protein expression could be biologically important in laryngeal tumorigenesis. Further research is required to clarify the involvement of ß-catenin in the mechanism associated with malignant transformation in laryngeal tissues.

Genetic and protein markers related to laryngeal epithelial precursor lesions and their neoplastic progression.

CONCLUSION: Various biomarkers might ultimately prove to have prognostic value and could be clinically relevant. It is mandatory confirm the prognostic power of these markers in large, well-designed, and prospective studies. OBJECTIVE: The aim of this study was to investigate the possible role of specific genes and proteins in laryngeal tumorigenesis. METHODS: Genetic analysis by multiple ligation-dependent probe amplification and analysis of protein expression by immunohistochemistry were carried out in a series of 50 tissue samples. RESULTS: In the smoker normal mucosa group TP53 loss was predominant, whereas in the epithelial precursor lesions (EPLs) CDKN2A loss and BCL2L1 gain were most frequent. EPL with progression presented CTNNB1 loss. Positivity at cytoplasm for ß-catenin, cyclin D1 and p53 was detected in all EPL cases with progression to invasive carcinoma. Multivariate analysis showed that expression of ß-catenin and loss of CTTNB1 were associated with laryngeal cancer risk.

Evidence of nestin-positive cells in the human cutaneus Meissner and Pacinian corpuscles.

Nestin is an intermediate filament protein expressed in neuroepithelial stem cells during development and it is later replaced by cell specific neuronal or glial filaments. Nevertheless, nestin⁺ cells remain within adult tissues and they can be regarded as potential neural stem cell (NSC). Nestin⁺ cells have been detected in Schwann cells related with sensory corpuscles of rodent and they have been demonstrated to be NSC. We have investigated the existence of nestin⁺ in human cutaneous cells Meissner and Pacinian corpuscles through the use of immunohistochemistry techniques and in situ hybridization. S100 protein (also regarded as a marker for NSC) and vimentin (the intermediate filament of mature Schwann cells in sensory corpuscles) were also investigated. The results show that the adult human cutaneous sensory Meissner and Pacinian corpuscles contains a small population of Schwann-related cells (vimentin⁺) which on the basis of their basic immunohistochemical characteristics (S100 protein⁺, nestin⁺) can be potential NSCs. Cells sharing identical immunohistochemical profile were also found in the close vicinity of Meissner corpuscles. Because their localization they are easily accessible and may represent a peripheral niche of NSC to be used for therapeutic goals.

Análisis de la inestabilidad de microsatélites en el carcinoma escamoso de laringe / Analysis of microsatellite instability in laryngeal squamous cell carcinoma

Introducción y objetivos: La literatura sobre la participación de la inestabilidad de microsatélites en el carcinoma de células escamosas de cabeza y cuello muestra una gran variabilidad, probablemente debido a las diferencias en la metodología de las pruebas. Utilizando un sistema de detección consensuado, nos planteamos como objetivo llegar a una estimación fiable de la prevalencia de la inestabilidad de microsatélites en un subconjunto de carcinomas de células escamosas de cabeza y cuello. Métodos: Se analizó el estado de inestabilidad de microsatélites en 43 pacientes no tratados previamente y diagnosticados de un carcinoma primario de células escamosas de laringe mediante una prueba de PCR múltiple, incluyendo 5 marcadores repetidos de mononucleótidos. Resultados: En 36 casos se observó un fenotipo estable o microsatélites estables (83,7%), y en 7 casos (16,3%) se mostró un fenotipo positivo de inestabilidad de microsatélites. Uno de los casos mostró inestabilidad en 3 de los 5 marcadores, otro mostró inestabilidad en 2 marcadores y 5 casos en un marcador. Entre los casos de inestabilidad de microsatélites positiva y los casos estables no hubo diferencias con respecto a la edad, el estadio del tumor, la afectación de los ganglios linfáticos o las metástasis a distancia. Conclusiones: Nuestros datos muestran que una parte de los carcinomas de células escamosas de laringe presentan inestabilidad de microsatélites. El conocimiento sobre el estado de inestabilidad de microsatélites de los pacientes permitirá el ajuste de la terapia anti-cancerígena a nivel individual(AU)

Introduction and objectives: The literature on the involvement of microsatellite instability in head and neck squamous cell carcinoma shows great variability, probably due to differences in the testing methods. Using a consensus detection system, we aimed to reach a reliable estimate of microsatellite instability prevalence in a subset of head and neck squamous cell carcinoma cases. Methods: The microsatellite instabilityI status of 43 patients with previously untreated primary laryngeal squamous cell carcinomas was analyzed by a multiplex polymerase chain reaction assay including 5 mononucleotide repeat markers. Results: Thirty-six cases showed a stable phenotype or a microsatellite stable phenotype (83.7%) and 7 cases (16.3%) showed an microsatellite instability-positive phenotype. One case showed instability in 3 of 5 markers, 1 case in 2 markers and 5 cases in 1 marker. The microsatellite instability-positive and stable cases did not differ with respect to age, tumour stage, lymph node or distant metastases. Conclusions: Our data showed that a proportion of laryngeal squamous cell carcinomas are microsatellite instability positive. Knowledge of microsatellite instability patient status will allow adjusting anticancer therapy at an individual level(AU)

[Analysis of microsatellite instability in laryngeal squamous cell carcinoma]. / Análisis de la inestabilidad de microsatélites en el carcinoma escamoso de laringe.

INTRODUCTION AND OBJECTIVES: The literature on the involvement of microsatellite instability in head and neck squamous cell carcinoma shows great variability, probably due to differences in the testing methods. Using a consensus detection system, we aimed to reach a reliable estimate of microsatellite instability prevalence in a subset of head and neck squamous cell carcinoma cases. METHODS: The microsatellite instabilityI status of 43 patients with previously untreated primary laryngeal squamous cell carcinomas was analyzed by a multiplex polymerase chain reaction assay including 5 mononucleotide repeat markers. RESULTS: Thirty-six cases showed a stable phenotype or a microsatellite stable phenotype (83.7%) and 7 cases (16.3%) showed an microsatellite instability-positive phenotype. One case showed instability in 3 of 5 markers, 1 case in 2 markers and 5 cases in 1 marker. The microsatellite instability-positive and stable cases did not differ with respect to age, tumour stage, lymph node or distant metastases. CONCLUSIONS: Our data showed that a proportion of laryngeal squamous cell carcinomas are microsatellite instability positive. Knowledge of microsatellite instability patient status will allow adjusting anticancer therapy at an individual level.

Genomic profiling of sinonasal squamous cell carcinoma.

BACKGROUND: Sinonasal squamous cell carcinomas (SCCs) are rare tumors with no etiologic link to tobacco and alcohol, as opposed to other SCCs of the head and neck (HNSCC). Little is known about the genetic changes in sinonasal SCC. METHODS: DNA copy number changes of sinonasal SCC were analyzed by multiplex ligation-dependent probe amplification (MLPA) and microarray comparative genomic hybridization (maCGH), and results were related to clinicopathologic features. RESULTS: Copy number losses most frequently included genes at 9p21, 13q14, 17p13, 17q21, and 18q11. Frequent gains were observed on 8q24, 11q13, 17q12, 19p13, and 20q11-q13. CONCLUSION: The genomic profile of sinonasal SCC showed a number of chromosomal regions with copy number changes similar to those known in HNSCC, in spite of the differences in etiology.

Genetic model of transformation and neoplastic progression in laryngeal epithelium.

BACKGROUND: The aim of this study was to analyze genetic alterations in the transformation-progression model of laryngeal tumors. METHODS: Copy number changes of 37 genes were analyzed by multiple ligation-dependent probe amplification (MLPA) in 94 tissue samples. RESULTS: In the smoker normal mucosa group TP53 loss was predominant, whereas in the precursor lesions CDKN2A loss and CDKN2D gain were most frequent. Precursor lesions with progression presented CTNNB1 loss. In the carcinoma group the most common changes were CDKN2A, MLH1, CTNNB1, and CASP6 losses and RECQL4, CCND1, and EMS1 gains. Positive lymph node primary tumors were related to TP53, IL1A, and RB1 losses and STK11 gain. The lymph node metastases differed from their corresponding primary tumor in LMNA, RECQL4, and IGF1R losses, and N33 and CDKN2D gains. CONCLUSIONS: Genetic changes and new key genes were found to be associated with specific steps. We included new steps, not presented in the classic models: normal mucosa tobacco exposed, positive lymph node primary tumor, and corresponding lymph node metastases.

Genetic differences between primary larynx and pharynx carcinomas and their matched lymph node metastases by multiplex ligation-dependent probe amplification.

Lymph nodes metastasis is a major risk factor related to poor survival in larynx and pharynx carcinomas. The aim of this study is to search for markers of lymph node involvement analyzing the genetic differences between primary larynx and pharynx squamous cell carcinomas and their corresponding lymph node metastases. Twenty-five primary tumors and their corresponding lymph node metastases were examined. DNA copy number changes of 37 genes were analyzed by multiplex ligation-dependent probe amplification (MLPA). Loss of CDKN2A (9p21) occurred in 14 out of 25 pairs (56%) of primary tumor and lymph node metastases. Loss of LMNA (1q21) was exclusively detected in 8 lymph node samples (32%). Loss of CTNNB1 (3p22) and gain of CDKN2D (19p13) were also significantly more frequent in lymph node metastases. Other aberrations related to lymph node metastases were loss of MFHAS1 (8p23), RECQL4 (8q24) and gain of N33 (8p22) and TP53 (17p13). Primary tumor and corresponding lymph node metastases showed common genetic changes. However, the lymph node metastases presented with a number of additional alterations. Acquisition of these alterations may play a role in lymphatic metastasis development.

[Nasosinusal adenocarcinoma: molecular and genetic analysis by MLPA]. / Análisis genético molecular con MLPA en los adenocarcinomas nasosinusales.

INTRODUCTION AND AIM: Intestinal-type sinonasal adenocarcinomas (ITACs) are rare epithelial tumours, primarily originating in the nasal cavities and paranasal sinuses and characterized by glandular structures. The aims of this study are: to determine the genetic alterations in ITACs by MLPA (Multiplex ligation-dependent probe amplification) and to correlate the findings to the clinical behavior and follow-up information of the patients. MATERIAL AND METHOD: We performed a longitudinal prospective study on 20 patients with ITAC, seen in our department between 1998 and 2004. DNA was extracted from primary tumor samples and analyzed by MLPA. RESULTS: The T stage of our series was T2: 4 (20 %), T3: 6 (30 %), T4a: 3 (15 %) and T4b: 7 (35 %). All cases initially were N0 and M0. Seventeen patients (85 %) had professional exposure to wood dust. All patients underwent surgical intervention and 70 % received complementary radiotherapy. Overall 5 and 10 year survival was 42 % and 22 %, respectively. Gains were found most frequently for PTP4A3 and PDCD8 (65 %), TNRFSF7 (50 %), RECQL4 and LMO2 (45 %), and losses for BCL2 (70 %), IL13 (55 %), ABCB1 and RB1 (50 %), PIK3CA and CDH1 (45 %). CONCLUSIONS: Losses of F3, MIF, and BRCA1 significantly correlated with the posterior development of metastases and with worse survival. Also gains of PIK3CA, UTY, and RELA correlated with poor clinical outcome. Losses of BRCA1 and F3 were significant in multivariate analysis.

Análisis genético molecular con MLPA en los adenocarcinomas nasosinusales / Nasosinusal adenocarcinoma: molecular and genetic analysis by MLPA

Introducción y objetivo: Los adenocarcinomas nasosinusales (ACNS) son tumores epiteliales raros, primarios de las fosas nasales y los senos paranasales, que se caracterizan por presentar estructuras glandulares. Los objetivos de este trabajo son: determinar las alteraciones génicas en los ACNS mediante MLPA (multiplex ligation-dependent probe amplification) y establecer la relación entre los cambios génicos del tumor y el comportamiento clínico-evolutivo de los pacientes. Material y método: Realizamos un estudio longitudinal prospectivo a 20 pacientes con ACNS controlados en nuestro servicio entre 1998 y 2004. A las muestras tumorales se les extrajo el ADN para realizar la MLPA. Resultados: La categoría T de los pacientes fue para T2: 4 (20 %), T3: 6 (30 %), T4a: 3 (15 %) y T4b: 7 (35 %). Todos eran inicialmente N0 y M0. El 85 % había estado expuesto, en el medio laboral, al polvo de madera. El tratamiento indicado fue quirúrgico (100 %) con radioterapia complementaria (70 %). La supervivencia global a los 5 y 10 años fue del 42 y el 22 %, respectivamente. Las ganancias génicas más frecuentes fueron: PTP4A3 y PDCD8 (65 %), TNRFSF7 (50 %), RECQL4 y LMO2 (45 %); las pérdidas: BCL2 (70 %), IL13 (55 %), ABCB1 y RB1 (50 %), PIK3CA y CDH1 (45 %). Conclusiones: La aparición de metástasis se correlacionó de forma significativa con pérdidas en F3, MIF y BRCA1. La peor supervivencia con pérdidas en F3, BCRA1 y MIF y ganancias en PIK3CA, UTY y RELA. En el análisis multivariable alcanzaron significación estadística las pérdidas en BRCA1 y F3

Introduction and aim: Intestinal-type sinonasal adenocarcinomas (ITACs) are rare epithelial tumours, primarily originating in the nasal cavities and paranasal sinuses and characterized by glandular structures. The aims of this study are: to determine the genetic alterations in ITACs by MLPA (Multiplex ligation-dependent probe amplification) and to correlate the findings to the clinical behavior and follow-up information of the patients. Material and method: We performed a longitudinal prospective study on 20 patients with ITAC, seen in our department between 1998 and 2004. DNA was extracted from primary tumor samples and analyzed by MLPA. Results: The T stage of our series was T2: 4 (20 %), T3: 6 (30 %), T4a: 3 (15 %) and T4b: 7 (35 %). All cases initially were N0 and M0. Seventeen patients (85 %) had professional exposure to wood dust. All patients underwent surgical intervention and 70 % received complementary radiotherapy. Overall 5 and 10 year survival was 42 % and 22 %, respectively. Gains were found most frequently for PTP4A3 and PDCD8 (65 %), TNRFSF7 (50 %), RECQL4 and LMO2 (45 %), and losses for BCL2 (70 %), IL13 (55 %), ABCB1 and RB1 (50 %), PIK3CA and CDH1 (45 %). Conclusions: Losses of F3, MIF, and BRCA1 significantly correlated with the posterior development of metastases and with worse survival. Also gains of PIK3CA, UTY, and RELA correlated with poor clinical outcome. Losses of BRCA1 and F3 were significant in multivariate analysis

[Tumour recurrence in squamous head and neck cancer]. / Recidivas en el cáncer escamoso de cabeza y cuello.

INTRODUCTION: For most patients with squamous head and neck cancer (HN-SCC), locoregional tumour recurrence (TR) carries an extremely poor prognosis and is therapeutically challenging. OBJECTIVE: To define the clinical aspects of TR and their implication on the survival in patients with HN-SCC. PATIENTS AND METHOD: The clinical management and the outcome of 652 patients with HN-SCC were reviewed. RESULTS: The overall incidence of TR in this series of HN-SCC was 19.9% (n=130). The most frequent locations of the primary cancers were oropharynx (32%), hypopharynx (24%), and oral cavity (21%). The rates of recurrence were locoregional 50%, local 43 % and stomal recurrence 7%. The appearance of a TR reduces the overall survival of patients with HN-SCC to 15%. Survival is better in glottic (38%) and supraglottic (27%), carcinomas, and worse in oro-hypopharynx tumours (2-4%). CONCLUSIONS: RT are more frequent in pharyngeal tumours, especially locoregional recurrences. Patients with recurrence in pharynx were definitely associated with poor prognosis and in these cases salvage surgery seems not to improve survival rates.

Recidivas en el cáncer escamoso de cabeza y cuello / Tumour recurrence in squamous head and neck cancer

Introducción: Las recidivas tumorales (RT) causan elevada mortalidad en los pacientes con cáncer escamoso de cabeza y cuello (CECC) y complican mucho la actitud terapéutica. Objetivo: Describir los aspectos clínicos de las RT y su implicación en la supervivencia. Pacientes y método: Se revisa a 652 pacientes con CECC y se recoge las características clínicas de las RT observadas. Resultados: Las RT se presentaron en el 19,9 % (130 pacientes) de nuestra serie de CECC. Las localizaciones más frecuentes son mesofaringe (32 %), hipofaringe (24 %) y cavidad oral (21 %). El 50 % de las RT son de carácter locorregional; el 43 %, locales, y el 7 %, estomales. La aparición de una RT reduce al 15 % la supervivencia general de los pacientes con CECC, es mayor en la laringe glótica (38 %) y la supraglótica (27 %), y baja al 2-4 % en mesofaringe e hipofaringe. Conclusiones: Las RT son más frecuentes en tumores faríngeos y de tipo locorregional. El pronóstico en esta localización es muy malo y el tratamiento de rescate apenas modifica la supervivencia

Introduction: For most patients with squamous head and neck cancer (HN-SCC), locoregional tumour recurrence (TR) carries an extremely poor prognosis and is therapeutically challenging. Objective: To define the clinical aspects of TR and their implication on the survival in patients with HN-SCC. Patients and method: The clinical management and the outcome of 652 patients with HN-SCC were reviewed. Results: The overall incidence of TR in this series of HN-SCC was 19.9 % (n=130). The most frequent locations of the primary cancers were oropharynx (32 %), hypopharynx (24 %), and oral cavity (21 %). The rates of recurrence were locoregional 50 %, local 43 % and stomal recurrence 7 %. The appearance of a TR reduces the overall survival of patients with HN-SCC to 15 %. Survival is better in glottic (38 %) and supraglottic (27 %), carcinomas, and worse in oro-hypopharynx tumours (2-4 %). Conclusions: RT are more frequent in pharyngeal tumours, especially locoregional recurrences. Patients with recurrence in pharynx were definitely associated with poor prognosis and in these cases salvage surgery seems not to improve survival rates

Chromosomal changes in relation to clinical outcome in larynx and pharynx squamous cell carcinoma.

Invasive head and neck squamous carcinomas are among the cytogenetically most complex tumors. Perhaps for this reason, there is little consensus on the prognostic value of specific chromosomal aberrations. Here we present results of CGH analysis of 56 clinically well-characterized set of head and neck cancers, consisting of larynx and pharynx only. The aim was to find possible associations with clinical outcome. The major chromosome arms showing gains were (in decreasing order): 3q, 7q, 8q, 5p, 11q13, 17q and 18p, and losses occurred at 3p, 11qter, 4p, 18q, and 5q. The segments most frequently amplified were 3q26-qter, 11q13, 11q22, 3q12-13, 18p11.3, 18q11.2 and 8q24.3. Tumors with stages III and IV, and lymph node positive tumors had a worse clinical outcome. Surprisingly, no specific chromosomal abnormality correlated with disease-free survival. The only aberration that correlated to one of the clinico-pathological parameters was amplification 11q13, that occurred solely in lymph node positive, stage IV tumors. However 11q13 amplification did not correlate with disease-free survival. These results seem to indicate that genetic alterations at the level of chromosomes have limited prognostic value in patients with invasive larynx and pharynx squamous cell carcinomas.

Numerical aberrations of chromosomes 8, 9, 11, and 17 in squamous cell carcinoma of the pharynx and larynx: a fluorescence in situ hybridization and DNA flow cytometric analysis of 50 cases.

Squamous cell carcinoma of the pharynx and larynx (SCCPL) is a genetically complex disease and is frequently associated with nonrandom chromosomal alterations. Fifty primary SCC of the pharynx (oropharynx, n=11): see and hypopharynx, n=11) and larynx ( n=28) were examined for numerical aberrations of chromosomes 8, 9, 11, and 17 with a panel of chromosome-specific repetitive DNA probes by fluorescence in situ hybridization (FISH). DNA ploidy analysis was also performed by flow cytometry (FCM). Aneusomic copy numbers of chromosomes 8, 9, 11, and 17 were discovered in 66%, 68%, 68% and 78% of tumors, respectively. FCM showed abnormal DNA content in 74% of cases (mean DNA index=1.69). Polysomy was the main finding in both DNA-aneuploid and DNA-diploid tumors (64.5% of cases). Numerical aberrations of chromosomes 8 and 11 correlated to DNA ploidy by FCM (P< 0.05). Aneusomy was present in 69.23% of DNA-diploid tumors. Marked intratumoral and intertumoral chromosomal heterogeneity was noted between individual tumors, suggesting a notable heterogeneity in aneuploid and diploid cell populations. Interphase FISH can be used to study important cytogenetic changes which occur during the development of SCC of the pharynx and larynx.