人乳头状瘤病毒阳性的头颈鳞状细胞癌特异性基因生物信息学研究

目的分析人乳头状瘤病毒(HPV)阳性的头颈鳞状细胞癌(鳞癌)特异表达基因及关键信号通路,为HPV相关头颈鳞癌筛选有价值的基因标记物,并为进一步的肿瘤机制研究提供参考。方法从GEO高通量基因芯片数据库中筛选出头颈鳞癌具有HPV感染信息的芯片,从中筛出差异基因进行基因本体分析及京都基因和基因组(KEGG)信号通路富集分析,并筛出头颈鳞癌的特征基因簇和通路,以及关键基因并进行蛋白质相互作用网络可视化分析。通过Cbioportal信息门户以及癌症基因组图谱(TCGA)数据库验证这些特异基因在HPV(+)与HPV(-)头颈鳞癌中的表达差异并分析特异基因与头颈鳞癌患者生存预后的相关性。结果从数据集GSE52088与GSE39366中筛选出42个共同差异基因,其中上调基因25个,下调基因17个,经Cytoscape两轮筛选确定白介素-6(IL-6)、细胞表面标记物CD44、基质金属蛋白酶1(MMP1)、CXC趋化因子配体基序1(CXCL1) 4个特异基因。信号通路富集分析显示共同差异基因参与细胞周期、NOD样受体信号通路、肿瘤坏死因子(TNF)信号通路途径等信号通路(P < 0.01)。经TCGA数据库以及Cbioportal检验证实特异基因在HPV(+)与HPV(-)头颈鳞癌中的表达差异,且IL-6、CD44表达水平与头颈鳞癌生存预后呈负相关(P < 0.01)。结论HPV(+)头颈鳞癌具有特异性基因表达,并可能参与关键信号通路调控肿瘤的发生发展。IL-6、CD44、MMP1、CXCL1 4个特异基因可能参与HPV(+)头颈鳞癌发展及侵袭过程,其中MMP1、CXCL1有望作为诊断及预后的标志物,IL-6、CD44与头颈鳞癌预后存在相关性,有望成为治疗HPV(+)头颈鳞癌的潜在靶点。     

Expression of type IV collagen and matrix metalloproteinase‐2 (type IV collagenase) in relation to nodal status in laryngeal cancer

Expression of type IV collagen and matrix metalloproteinase‐2 (type IV collagenase) in relation to nodal status in laryngeal cancer Laryngeal carcinoma has a lower incidence of neck metastases than other malignant carcinomas of the head and neck region. However, some cases are very aggressive, showing neck metastases even in the early stages. In this study the expression of collagen IV and type IV collagenase (MMP‐2) were examined immunohistologically in 50 patients with laryngeal carcinomas, and the results were compared with the incidence of neck metastases and other clinicopathological factors. The correlation between collagen IV expression and the existence of nodal metastases was statistically significant (P < 0.001). There was also significant correlation between collagen IV expression and the histological grading of the tumour. There was a tendency for samples with continuous collagen IV staining to have no matrix metalloproteinase‐2 (MMP‐2) immunoreactivity. No significant correlation was seen between MMP‐2 protein expression and clinicopathological parameters although the correlation between MMP‐2 and existence of nodal metastases was statistically borderline (P = 0.07). Multivariate analysis of the clinicopathological factors that may have an influence on the nodal status in laryngeal cancer revealed that, apart from T stage, collagen IV pattern in the basement membrane surrounding nests of carcinoma is an important prognostic factor.     

Serum levels of vascular endothelial growth factor in patients with head and neck cancer

Angiogenesis is now considered to be crucial for tumor growth and metastasis. In several tumors, microvascular density has been shown to be correlated with metastasis and aggressiveness. Vascular endothelial growth factor (VEGF) is a secreted endothelial cell-specific mitogen, which is induced by hypoxia and is angiogenic in vivo. VEGF has been identified in a wide variety of malignancies including head and neck squamous cell carcinomas (HNSCC). We investigated the circulating level of VEGF in sera from patients with various head and neck squamous cell carcinomas (n = 71) as well as from healthy normal controls (n = 47). Serum VEGF concentrations were determined as serum immunoreactivity by using a quantitative sandwich enzyme immunoassay technique. For statistical analysis, the Wilcoxon 2-sample test and Kruskal-Wallis test were performed. The majority of the patients with HNSCC were found to have high concentrations of serum VEGF. The levels of VEGF in the sera of patients with cancer ranged from below the detection limit to 937.1 pg/ml (mean, 144.5 pg/ml). In contrast, the VEGF serum levels in 47 healthy individuals ranged from below the detection limit to 168.1 pg/ml (mean, 32.7 pg/ml), VEGF serum concentration being significantly higher in HNSCC patients (P = < 0.001). These findings indicate that a positive angiogenesis regulator such as VEGF might function as an endocrine growth factor, particularly for solid HNSCC tumors and may be a useful marker for clinical monitoring. Received: 12 April 1999 / Accepted: 30 September 1999     

MASPIN tumour-suppressing activity in head and neck squamous cell carcinoma: Emerging evidence and therapeutic perspectives

Conclusions. A large series confirmation may suggest elective neck dissection in cN0 MASPIN-negative oral squamous cell carcinomas (SCCs). Re-activated nuclear MASPIN in association with anti-angiogenic or cytotoxic drugs may be extremely effective in the treatment of laryngeal SCC. Objectives. MASPIN is a serpin showing a unique tumour-suppressing activity. Despite the significant incidence of head and neck SCC (HNSCC), a limited number of studies has considered MASPIN's role in these malignancies. This review focuses on the attempts to translate MASPIN's HNSCC suppressive properties toward a diagnostic/prognostic tool and a novel carcinoma therapy. Methods. A systematic literature review was performed to investigate available evidence about MASPIN subcellular localization and biological role in HNSCC. Results. Five studies have investigated MASPIN's expression in oral SCC. Most of them concluded that low or absent MASPIN cytoplasmic expression was more frequent in oral carcinomas with lymph node metastases. Only our group evaluated MASPIN's role in laryngeal SCC. Higher nuclear MASPIN expression was found in patients without SCC recurrence and was significantly associated with longer disease-free survival. Micro-vascular density was lower in laryngeal carcinomas with MASPIN nuclear staining. The relation between MASPIN and M30-assessed apoptosis in laryngeal SCC supports the hypothesis of an important apoptosis-sensitizing effect of nuclear MASPIN.     

Differential expression of p53, p63 and p73 proteins in human buccal squamous‐cell carcinomas

Abnormalities in the p53 gene are regarded as the most consistent of the genetic abnormalities in oral squamous‐cell carcinoma. Two new members of the p53 gene family, p73 and p63, have recently been identified, with the three sharing considerable sequence homology at the acidic N‐terminal transactivation, central DNA‐binding and C‐terminal oligomerization domains, indicating possible functional and biological interactions. The differential expression of p73, p63 and p53 genes in human oral squamous‐cell carcinoma does not yet appear to be completely understood, however. In this study, therefore, immunohistochemical analysis of protein expression was performed for 40 samples of well‐differentiated human buccal squamous‐cell carcinomas, with 10 specimens of normal buccal mucosa employed as controls. Differential expressions of p63, p73 and p53 proteins in the carcinoma samples were: p63+/p73+/p53 + (n = 28; 70%); p63+/p73+/p53– (n = 4; 10%); p63+/p73–/p53– (n = 8; 20%), respectively; and p63+/p73+/p53– for normal mucosa (n = 10; 100%). A significant correlation between p53, p63 and p73 immunoexpression was demonstrated for the buccal squamous‐cell carcinoma samples (P < 0.0001; Fisher's exact test). Significance was not achieved for the correlation between p73 and p53 immunoexpression and clinicopathological parameters for buccal carcinomas (P > 0.05; Fisher's exact test). Our results indicate that both p73 and p63 may be involved in the development of human buccal squamous‐cell carcinoma, perhaps in concert with p53.     

Syndecan-1 expression in laryngeal cancer

Syndecan-1 is a member of the syndecan family of cell surface heparan sulphate proteoglycans. The aim of this study was the evaluation of syndecan-1 expression in laryngeal cancer. The findings were correlated both with the clinico-pathological parameters of the tumours and with patient survival. Paraffin-embedded samples from 48 patients with laryngeal cancer selected from the files of the ENT Department of the Medical Academy in Lublin were immunostained with anti-syndecan-1 monoclonal antibody. The patients’ mean age was 56 years, and 69% survived for over 3 years . Syndecan-1 immunoreactivity was observed in 48 (100%) of carcinomas. In our study, statistically significant correlations were observed both between syndecan-1 expression and patient survival (Chi-square = 4.364; P<0.05) and between syndecan-1 expression and clinical stage of disease (Chi-square = 4.363; P<0.05). Significant differences in syndecan-1 expression were also observed with various stages of histological differentiation of the carcinomas (Chi-square = 6.588; P<0.05), and also according to the presence or absence of metastatic changes in the regional lymph nodes (Chi-square = 6.289; P<0.05). Our results indicate that syndecan-1 could be used as a prognostic marker in laryngeal cancer. Accepted: 3 October 2001     

Snail as a novel marker for regional metastasis in head and neck squamous cell carcinoma

Objective

Previous studies have shown Snail expression integral to the epithelial-mesenchymal transition during tumor progression. However, its behavior in clinical head and neck squamous cell carcinomas (HNSCCs) is yet undefined. We therefore sought to (1) investigate clinical and histopathologic characteristics of Snail-positive HNSCC and (2) understand the link between Snail and other commonly used HNSCC tumor markers.

Study Design

A retrospective case-control study was conducted.

Setting

This study was conducted in a large-scale academic center.

Study Subjects

Of 51 consecutive HNSCC, 42 surgical resections were included.

Methods

Two separate pathologists performed standard histopathologic reviews along with immunohistochemistries (Snail, E-cadherin, p16, epidermal growth factor receptor [EGFR]) and in situ hybridization (human papilloma virus [HPV]). Medical review for all cases was performed.

Results

Twenty-two (52%) of 42 cases stained 4+ Snail (>75% staining). The remaining 20 cases were considered negative. Snail was strongly inversely related to E-cadherin expression (ρ = −0.69, P < .001), but statistically independent from HPV, p16, or EGFR expression. Snail(+) tumors were equally represented from each anatomic subsite. Snail(+) tumors were strongly associated with poor differentiation (P < .001) and basaloid classification (P = .004). Snail(+) tumors were also strongly associated with lymphovascular invasion (P = .02), but not perineural invasion. Ultimately, 11 (50%) of 22 of Snail(+) tumors demonstrated positive nodal metastasis and 11 (79%) of 14 node-positive cases were Snail(+) (P = .02).

Conclusion

This pilot study provides promising evidence of Snail' role as a molecular prognostic marker for HNSCC. Snail positivity is significantly predictive of poorly differentiated, lymphovascular invasive, as well as regionally metastatic tumors. Because Snail positivity appears independent of HPV, p16, and EGFR expression, Snail may prove to improve upon these markers' predictive limitations.     

Bcl‐Xl protein expression in laryngeal squamous cell carcinoma

Bcl‐X1 protein expression in laryngeal squamous cell carcinoma The Bcl‐2 family of proteins regulate one of the steps in an evolutionary conserved apoptotic pathway. The long splice variant of Bcl‐X (Bcl‐Xl) is a potent antagonist of apoptosis. The aim of the study was to evaluate the relation between the presence of immunohistochemically detectable Bcl‐Xl protein in laryngeal squamous cell carcinomas (LSCCs) and clinicopathological data, as well as DNA ploidy status and proliferative activity. In 50 specimens of LSCC, Bcl‐Xl protein expression was evaluated immunohistochemically. Proliferative activity (SG₂M‐phase index) and DNA ploidy were measured by flow cytometry. In our study, Bcl‐Xl protein expression decreased with decreasing tumour differentiation (P = 0.04). The majority of patients with Bcl‐Xl protein immunoreactivity had no metastatic lymph node involvement (P = 0.01). Other factors such as age, gender, primary tumour size (pT) and type of cancer (keratinizing/non‐keratinizing) were not associated with Bcl‐Xl protein level. There was no correlation between Bcl‐Xl protein and SG₂M‐phase index or DNA ploidy status. Our findings show that expression of Bcl‐Xl protein is increased in a great fraction of laryngeal cancers. Further studies, however, are needed to clarify association between Bcl‐Xl protein expression and clinical course of patients.     

Prognostic significance of Ki-67 expression for patients with laryngeal squamous cell carcinoma primarily treated by total laryngectomy

The aim of this study was to determine the prognostic value of Ki-67 immunostaining in laryngeal squamous cell carcinomas. Ki-67 labeling was quantified in 63 laryngeal squamous cell carcinomas by counting at least 1,000 tumor cells in the most immunoreactive area in each sample, and the Ki-67 labeling index was calculated as a percentage. The antigen expression was compared with clinical factors, histopathological grading and prognosis. The Ki-67 mean proliferation index for all patients was 25.44% ( range, 2–75%). A significant correlation was found between Ki-67 mean proliferation index and patient age (P<0.05), T-stage (P<0.05), nodal metastasis (P=0.001) and recurrence (P<0.001). There was no significant association between the Ki-67 mean proliferation index and tumor site or histologic grade. A univariate analysis showed that the Ki-67 labeling index >21% (P<0.001), T-stage (P<0.001) and nodal metastasis (P=0.001) are determinants of recurrence. In the multivariate analysis, the Ki-67 labeling index >21% (P<0.001), T-stage (P<0.001) and nodal metastasis (P<0.05) were independent predictors of recurrence. Kaplan-Meier plots of survival in patients with Ki-67 values above and below the median (21%) of the general study population showed that a high Ki-67 labeling index correlated with a shorter disease-free survival (P<0.0001). The analysis of the Ki-67 labeling index at the time of initial surgery may be a powerful prognostic marker for patients with laryngeal squamous cell carcinoma and may be useful for selecting subgroups of patients who should be treated with more aggressive therapies.     

Identification of Genes Overexpressed in Head and Neck Squamous Cell Carcinoma Using a Combination of Complementary DNA Subtraction and Microarray Analysis

Objectives/Hypothesis To discover unique genes specific for squamous cell carcinoma of the head and neck for eventual development as tumor markers and vaccine candidates. Study Design Molecular biological analysis of fresh‐frozen head and neck squamous cell cancer (HNSCC). Methods A subtractive library was made from two HNSCC and six normal tissues using a polymerase chain reaction (PCR)–based approach. Genes from this library were PCR amplified and placed on a microarray glass slide. RNA was prepared or obtained from 16 fresh‐frozen HNSCC and 22 normal tissue sources. Fluorescent probes were made from the polyA+ RNA derived from the tumor and normal tissues. The probes were hybridized to the glass slides and excited by a tuneable laser. One hundred seven of the genes showing the highest differential fluorescence value between tumor and normal tissue were identified by sequence analysis. Results Thirteen independent genes were found to be overexpressed in tumor tissues. Of these, nine were previously known: keratins K6 and K16, laminin‐5, plakophilin‐1, matrix metalloproteinase‐2 (MMP), vascular endothelial growth factor, connexin 26, 14–3‐3 sigma, and CaN19. The level of polyA+ RNA of these genes in the tumors was significantly different from the levels in normal tissue (P < .05). Four previously unidentified genes were also discovered to have increased expression in tumor tissue. Comparing the total tumor group (n = 16) to the normal group (n = 22), only one of these genes showed significant overexpression. Conclusion We report the identification of nine known genes that are significantly overexpressed in HNSCC as compared to normal tissue using subtractive and microarray technology. In addition, we present four previously unidentified genes that are overexpressed in a subset of tumors. These genes will be developed as tumor markers and vaccine candidates.     

Argyrophilic nucleolar organizer regions as a prognostic indicator of laryngeal carcinomas

Nucleolar organizer regions (NORs) were studied in surgically removed specimens from 76 patients with squamous cell carcinomas of the larynx using an argyrophilic (Ag) staining technique. The mean number of AgNORs per nucleus was 4.3±1.38 (SD). The mean AgNOR number for T1 or T2 disease was statistically lower than that for T3 or T4 lesions (P<0.05). The mean AgNOR number was lower in patients with N0 disease than in patients with N1 or N2 or N3 tumors (P<0.05). There was also a statistically significant difference between the mean AgNOR number for stage II and stage III disease (P<0.01), for stage III and stage IV disease (P<0.05), but not for stage I and stage II disease. According to the histological grading, there was a significant difference between the mean AgNOR number for the well-differentiated and moderately differentiated tumors (P<0.05), and for the moderately differentiated and poorly differentiated types (P<0.01). These results suggest that the number of AgNORs in the nucleus is a significant indicator of laryngeal carcinomas.     

Nitric oxide and apoptosis during human head and neck squamous cell carcinoma development.

PURPOSE: Apoptosis index (AI), Bcl-2, and Bax have shown prognostic significance in head and neck squamous cell carcinoma (HNSCCa). Other areas of research have implicated nitric oxide (NO) or its various intermediate species in both proapoptotic and antiapoptotic processes. We have previously shown that NO-generating enzymes are significantly increased during the stepwise progression to HNSCCa. The aim of this study was to explore the interrelationship of NO and a known consequence of NO-related oxidative stress, apoptosis, during this step-wise process. MATERIALS AND METHODS: Formalin fixed-paraffin embedded tissue samples of 10 normal oral mucosa, 15 reactive/dysplastic lesions, and 17 HNSCCa lesions studied previously were subjected to the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP labeling (TUNEL) assay as well as immunohistochemical staining against Bcl-2, Bax, and p53. Patient charts were reviewed and clinical data were compared. The study pathologist (G.K.H) reviewed these slides blinded to patient identifiers or clinical data. The number of immunopositive cell nuclei or staining intensity was graded, noting the pattern of immunostaining. These staining characteristics were compared with the results of immunostaining previously obtained for endothelial constitutive NO synthase (ecNOS) and nitrotyrosine. RESULTS: Compared with normal oral mucosa, the AI, Bcl-2, Bax, Bcl-2/Bax intensity and frequency ratios, and mutant p53 intensity significantly changed in reactive/dysplastic and HNSCCa lesions (P <.001 for all). Correlations between the staining characteristics of the antigens studied are presented. Furthermore, perilesional inflammatory cells showed staining in the TUNEL assay. CONCLUSIONS: In a set of tissue samples previously well characterized, these new findings implicate a link between NO and the induction of apoptotic cell death in HNSCCa development.     

The expression of vascular endothelial growth factor (VEGF) and VEGF‐C in early laryngeal cancer: relationship with radioresistance

The expression of vascular endothelial growth factor (VEGF) and VEGF‐C in early laryngeal cancer: relationship with radioresistance Angiogenesis is essential for tumour growth and invasion. Vascular endothelial growth factor (VEGF) is a prime mediator of tumour angiogenesis. VEGF‐C is a closely related protein that effects lymphatic endothelial cells and may be important in the process of lymphatic metastasis. The purpose of this study was to evaluate the expression of these cytokines in patients with T1 and T2a glottic, squamous cell carcinoma, in comparison with normal epithelial control tissue, to ascertain any association with radioresistance. Twenty‐two tumours treated by radiotherapy (13 radiosensitive, nine radioresistant) and seven normal control tissues were studied. The minimum follow‐up was 2 years after radiotherapy. Expression of VEGF and VEGF‐C was evaluated by immunohistochemistry of formalin‐fixed, paraffin‐embedded biopsy specimens. Analysis was carried out using a quantitative computer image analyser. Both VEGF and VEGF‐C were detectable in tumour and normal control specimens. There was increased expression in tumour specimens of both VEGF (P = 0.03) and VEGF‐C (P < 0.001). In addition, the expression of VEGF‐C was associated with tumours of higher histological grade (P = 0.021). There was, however, no difference in VEGF and VEGF‐C expression between radioresistant and radiosensitive tumours. The expression of VEGF and VEGF‐C is increased in early laryngeal squamous cell carcinoma (SCC). However, measuring the expression of these proteins cannot predict radioresistance in this tumour group.     

Long‐term quality‐of‐life outcomes in children undergoing adenotonsillectomy for obstructive sleep apnoea: a longitudinal study

Clin. Otolaryngol. 2011, 36 , 475–481 Objectives: To assess a cohort of patients who underwent adenotonsillectomy for obstructive sleep apnoea, 4 years after surgery for evidence of continued and long‐term improvement in quality of life. We also sought to compare our results to Child Health Questionnaire scores obtained from our previous study. We also compared our data with a healthy UK children population from normative data available. Design: Longitudinal study. Settings: University Hospital Tertiary Referral Centre. Participants: A 4‐year follow‐up study of 37 children who underwent adenotonsillectomy for obstructive sleep apnoea confirmed on polysomnography. There were 19 boys and 18 girls from our initial cohort. The primary caregiver completed the validated Child Health Questionnaire Parental Form version‐28, 4 years after initial surgery. Our control group consist of 221 healthy children aged 6–18 that were included as ‘normal’ controls in a study looking at children with juvenile arthritis. The children were defined as healthy by a physician and/or after declaration by the parent. Main outcome measure: Child Health Questionnaire Parental Form version‐28 scores. Results: A total of 33 patients (89%) from our initial cohort were contacted. The mean age was 10.6 (median, 11; range, 5–16). When compared with results obtained 3 months postoperatively, the mean scores were higher in five domains and were statistically significant in three subscales (Role Limitations P < 0.00001; Bodily Pain P < 0.002; and Global Health P < 0.02). There was a significant deterioration in Behaviour subscale (P < 0.0007) in spite of surgery. Compared with controls, 4‐year follow‐up scores were higher in five domains with the Global Health domain (P < 0.0004) being statistically significant. When the 4‐year follow‐up scores were compared with preoperative values, these were higher in all 13 domains with statistically significant improvements in nine domains, indicating that improvements had persisted 4 years after surgery. At 4 years, however, the means scores in many domains remain lower when compared with controls. Conclusion: Quality‐of‐life data are an important measure when deciding on a specific clinical intervention. In the short term, quality‐of‐life measures have been shown to improve after adenotonsillectomy for obstructive sleep apnoea. Our study demonstrates that the benefits of surgery are still persistent and the children continue to improve in the long term.     

Prevalence and clinical features of human papillomavirus in head and neck squamous cell carcinoma in Okinawa,southern Japan

Previous studies from Okinawa, a subtropical island in southern Japan, demonstrated a higher prevalence of human papillomavirus (HPV) in oral carcinoma and a higher incidence of oral and pharyngeal carcinoma than those for mainland Japan. The present study aims to investigate epidemiologic and clinical features of HPV in head and neck squamous cell carcinoma (HNSCC) in Okinawa. A total of 150 DNA samples from 150 Okinawan patients with head and neck squamous cell carcinoma (HNSCC) were screened for HPV sequences by PCR using three consensus primer sets, and HPV types were determined by direct sequencing. The samples were consisted of 46 cases from the hypopharynx, 44 from the oropharynx, 16 from the larynx, 25 from the oral cavity, 10 from the maxillary sinus, and 9 from the nasopharynx. HPV DNA was detected in 45 (30.0%) HNSCCs, and HPV-16 was identified in 86.7% of positive specimens. The highest prevalence of the HPV sequence was found in oropharyngeal carcinomas (50.0%), especially in tonsillar cancer (63.6%). Multivariate analysis showed that oropharyngeal carcinoma (P = 0.002; OR = 5.34; 95% CI = 1.83–15.58), oral cavity carcinoma (P = 0.012; OR = 4.94; 95% CI = 1.43–17.10), and histological poor differentiation (P = 0.011; OR = 4.25; 95% CI = 1.39–13.04) each independently increased the prevalence of HPV infection. The present study reveals that patients with HNSCC, e.g., oropharyngeal and oral cavity carcinomas, in Okinawa have relatively high HPV-16 positive rates and low HPV-18 positive rates comparing with mainland Japan.     

Vascular endothelial growth factor in nasal polyps: a comparison of asthmatic and non‐asthmatic patients

The cause of nasal polyps remains unknown, although there is a well‐recognized clinical association between nasal polyposis and asthma. The characteristic histological features of nasal polyps include large quantities of extracellular fluid. Vascular endothelial growth factor (VEGF) is a potent mediator of angiogenesis and vascular permeability. This study aimed to compare expression of VEGF in nasal polyps from patients with asthma and those with no apparent respiratory disease. Twenty‐four asthmatic and 35 non‐asthmatic patients were studied using immunohistochemistry for VEGF. VEGF expression was identified in endothelial, inflammatory and epithelial cells. There was significantly greater endothelial expression of VEGF in asthmatic patients (P < 0.05). Greater epithelial expression was observed in asthmatic patients but this did not reach statistical significance (P = 0.07). There was no difference in the density of inflammatory cells expressing VEGF. Differences between the two groups may reflect differences in disease severity or in the nature of the inflammatory process.     

Foreign Body in the Peripheral Bronchus: Extraction Using an Interventional Radiologic Method

Conclusions. Inter-regional differences in the distribution of genetic polymorphisms in glutathione S-transferases (GSTs) exist, which may have significant effect on the outcome of other GST polymorphism studies. The GSTT1 null genotype appears to be involved in modulation of the risk for head and neck squamous cell carcinoma (HNSCC). Background. The risk of HNSCC is strongly associated with smoking of cigarettes and consumption of alcohol, resulting in a load of toxins/carcinogens. Detoxification of such exogenous harmful compounds often occurs by phase II enzymes such as GSTs. Proper functioning of these enzymes may be deficient due to the presence of particular genetic polymorphisms in these GSTs, and this may increase the risk for HNSCC. We compared the GSTT1, GSTM1 and GSTP1 genotype frequencies in two groups of healthy blood donors, collected from different but adjacent regions in the Netherlands, with those of a group of patients with HNSCC. Subjects and methods. The GSTM1,GSTT1 and GSTP1 genotype frequencies in two Dutch Caucasian control populations (n=207 and n=285) from different but adjacent geographical regions (Maastricht and Nijmegen; distance, 125 km) and 185 patients with HNSCC from the Maastricht region were determined by PCR-related methods. Results. For the occurrence of the GSTT1 null genotype we found a significant difference (p=0.003) between the two control groups (20.3% vs 33.0% null genotype in the Nijmegen and Maastricht control groups, respectively). Since the HNSCC patients were collected from the Maastricht area, comparison with the Maastricht controls reveals a significant difference for GSTT1 null rates, which are lower in patients vs controls (OR?=?0.49, CI?=?0.32–0.76).     

Serum vascular endothelial growth factor in patients with head and neck squamous cell carcinoma

Vascular endothelial growth factor (VEGF) is a key pro-angiogenic cytokine expressed by most human tumours. Two isoforms, VEGF₁₂₁ and VEGF₁₆₅, are soluble and can be assayed in serum. Serum VEGF has been shown to be significantly raised in patients with solid tumours and shows some promise as a potentially useful tumour marker. Serum levels of VEGF were assayed in 52 patients with untreated head and neck squamous cell carcinoma (HNSCC) and 104 healthy controls. Serum VEGF is significantly raised in patients with HNSCC (P < 0.001), but there was no association with either tumour stage or specifically the presence of nodal metastases. Sixteen patients (31%) had a higher serum VEGF than 95th centile of controls, suggesting that serum VEGF measurement is of little practical use as an initial diagnostic tool. The finding that patients with HNSCC have significantly raised serum VEGF probably relates to enhanced platelet aggregation in these patients.     

Very long‐term Voice Handicap Index Voice Outcomes after Montgomery Thyroplasty: A cross‐sectional study

Objective

The aim of this multicentric cross‐sectional study was to examine the permanency of Montgomery thyroplasty (MTIS) results from a patient's perspective.

Design

The study consisted of collecting Voice Handicap Index (VHI‐30) questionnaires from patients who had previously been operated with MTIS between 2 and 12 years before. Very long‐term (>2 years) postoperative data were compared with the previously acquired preoperative and early postoperative VHI results. Influence of factors such as age, gender, size/side of the prosthesis and length of the follow‐up were also analysed.

Setting

Multicentric study involving three tertiary European voice centres.

Participants

Forty‐nine unilateral vocal fold paralysis (UVFP) patients, treated by MTIS, were included in the study.

Main outcome measures

The Voice Handicap Index‐30 score.

Results & Conclusions

The median VHI was significantly different over time‐points (Friedman's test P < .001), with a significant difference between preoperative and early postoperative time‐points (median VHI: 70 vs 21, respectively; P < .001) and between preoperative and very long‐term postoperative time‐points (median VHI: 70 vs 16, respectively; P < .001). The median VHI did not differ for the early and very long‐term postoperative time‐points (median VHI: 21 vs 16; P = .470). Age differences, gender differences and size/side differences of the prostheses, centres where surgery took place and length of the follow‐up showed no significant influence. Medialisation thyroplasty (MT) overall and MTIS, in particular, should be considered as a possible standard of care for UVFP when permanency of voice results is sought.     

Expression of epidermal growth factor receptor in glottic carcinoma and its relation to recurrence after radiotherapy

expression of EGFR was determined immunohistochemically in two groups of patients with glottic carcinoma, one that recurred after a full course of radiotherapy and one that did not. Using a 4-graded scale (-, +,++,+++) 80% (12/15) of the recurrent carcinomas had a staining intensity and proportion of stained cells of ++ or more. The same figure for non-recurrent carcinomas was 39% (7/18). The difference is statistically significant (chi-squared with Yates' correction, P &<5). The results indicate that an increased expression of EGFR may influence the rate of recurrence of glottic squamous cell carcinoma after radiotherapy.