Hideki Naganishi

Endoscopic investigation and evaluation of anxiety for the management of globus sensation

OBJECTIVE To verify whether the severity of globus sensation would be affected by the results of investigations on possible underlying serious pathology in the head, neck and esophagus. METHODS Thirty-six patients with globus sensation were enrolled in this study. All the patients suffered relatively persistent globus sensation which remained after conventional laryngoscopy at their family doctors. These patients were investigated for possible underlying oropharyngeal and esophageal lesions using fiberoptic endoscopy. The severity of globus was evaluated by the visual analog scales (VAS), and the degree of anxiety was evaluated by the state section of State Trait Anxiety Inventory (STAI-s). The questionnaires were administered at their first visits, and 9.6±3.2 months after endoscopy. The follow-up data obtained from 22 patients were incorporated in the further evaluations. Multiple regression analysis was used to evaluate the relationship between the improvement of VAS scores and that of STAI-s scores. Afterwards, Pearson product-moment correlation coefficient was measured. The recorded images of fiberoptic endoscopy were retrospectively verified by an expert of upper gastrointestinal endoscopy. RESULTS No malignancies were observed in the endoscopic examination. Despite no treatment administered during the follow-up period, significant improvement of VAS scores was observed from the initial scores (40±21) to follow-up scores (27±27, p=0.014) in the patients examined in this study. The multiple linear regression analysis proved that the improvement of STAI-s scores was the only factor significantly affected the improvement of VAS scores (p=0.029) among the dependent valuables. The retrospective evaluation of the recorded images revealed comorbid esophagitis in 10 out of the 22 patients. When patients were stratified with the presence of comorbid esophagitis, significant improvement of VAS scores was observed only in the group without comorbid esophagitis at their follow up (17±20, p=0.026) compared with their initial scores (36±17). The multiple linear regression analysis proved that the improvement of VAS scores was significantly affected by the improvement of STAI-s scores (p=0.047) in this group. Moreover, significant positive relationship between the improvement of VAS scores and that of STAI-s scores was observed only in the group without comorbid esophagitis (r=0.61, p=0.047). CONCLUSION Proper investigation to prove no underlying serious pathology may lead to the improvement of globus sensation in the patients without comorbid esophagitis through the reduction of their anxiety even when their symptoms are relatively persistent. Our results also indicated that some treatments against esophagitis may be helpful for the improvement of globus sensation in the patients with this comorbid disease.

Abstract 4039: miR-196a is a novel diagnostic biomarker and therapeutic target for laryngeal cancer

Head and neck cancer (HNC) is a major cause of cancer-related mortality and morbidity, comprising roughly 6% of all malignancies worldwide, and laryngeal cancer has the highest incidence in it. At present, laryngeal cancer is treated with surgery and/or (chemo)radiation, each of which can have devastating consequences on speech and swallowing function. Even with the combined treatment approaches as mentioned above, laryngeal cancers with advanced stages have poor prognosis, therefore in need of novel less invasive treatments against their high morbidity. However, although to date its relevance to smoking has been well documented, the molecular mechanism involved in laryngeal cancer development and its highly sensitive biomarkers remain to be known. Thus, we have focused on microRNAs (miRNAs) showing highly tissue-or disease-specific patterns as potential novel biomarkers or therapeutic targets with clinical applicability. In this study, to screen the miRNAs highly sensitive to laryngeal cancer as putative biomarkers, we first examined the expression of 723 human miRNAs in 8 laryngeal tissues (1 polyp, 2 dysplasias, 3 laryngeal cancers and 2 neighboring normal tissues matched to laryngeal cancers) using microarrays (Agilent Human miRNA V2). The result showed the up-regulation of 6 miRNAs and the down-regulation of 3 miRNAs in laryngeal cancer compared to others. Subsequently, to confirm these findings and quantitate cancer-specific miRNAs, we performed quantitative RT-PCR (qRT-PCR) analysis (TaqMan miRNA assays, Applied Biosystems) using 48 laryngeal tissue samples including 16 cancers on 5 miRNAs (miR-196a, miR-130b*, miR-133b, miR-375, miR-455-5p), which showed markedly differential expression in cancers compared with others on microarrays. The results were consistent with those from microarrays, especially when cancer samples were compared with matched laryngeal tissues as normal controls. Of the 5 miRNAs examined, miR-196a showed significant increase in cancers when compared to either benign tissues or dysplasias. Next, to examine the utility of miR-196a as a promising biomarker for laryngeal cancer, we performed qRT-PCR on miR-196a using 83 tissue samples and 68 serum samples and found that miR-196a could be very useful in diagnosing the disease. We also analyzed miR-196a expression in FFPE sections by in situ hybridization and the result showed its robust expression in cancers but barely detectable in benign tissues Finally, to explore whether miR-196a can promote tumor growth in vitro or in vivo, we treated human laryngeal cancer cells or xenografted nude mice, respectively, with either miR-196a inhibitor or negative control inhibitor, and found that miR-196a inhibition suppresses tumorigenic properties of laryngeal cancer. Together with these findings, our study provided the first demonstration of miR-196a as a putative diagnostic biomarker and therapeutic target for laryngeal cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4039.

MicroRNA Expression Signature in Laryngeal Cancer

OBJECTIVES: In this study, we examined the microRNA (miRNA) expression profile in laryngeal cancer. We further investigated the role of miR-133b and miR-375 in the growth of head and neck cancer (HNC) in vitro. METHODS: Expression profile of 723 human miRNAs in 8 laryngeal tissues (polyp, dysplasia, cancer, neighboring normal tissue matched to laryngeal cancer) was examined using microarrays. Subsequently, qRT-PCR was performed to confirm the microarray data using 48 laryngeal tissues including 16 laryngeal cancers on 3 miRNAs (miR-130b*, miR-133b, miR375), which showed distinctive expression in cancer on microarray. Six cancer samples had matched laryngeal tissues next to the tumor. Pairwise Mann-Whitney’s U test and Wilcoxon signed ranks test were utilized for statistical analyses. Furthermore, impact of miR-133b or miR-375 transfection on the growth of HNC cell lines was examined using WST-based assay. RESULTS: Microarray analysis revealed the up-regulation of 8 miRNAs, including miR-130b* and the down-regulation of 3 miRNAs, including miR-133b and miR-375. The results obtained from qRT-PCR were consistent with those obtained from microarray. Especially, the difference of miR-133b expression level between matched samples was significant (p 0.0277). MiR-133b suppressed the growth of SAS cell line and miR-375 suppressed the growth of JHU-011 cell line 5 days after transfection compared to control. CONCLUSIONS: We demonstrated the miRNA expression signature in laryngeal cancer and the role of miRNAs on the growth of HNC cell lines. Our novel findings may pave the way to elucidate the molecular mechanism of oncogenesis in HNC.

S192 – 3D Diagnosis of Vocalizing Larynx Using Cone Beam CT

Objectives Cone beam computed tomography (CBCT) was developed for the office-based quick and precise 3D visualization of maxillofacial region. CBCT requires scanning time of less than 10 seconds and provides isotropic 3D image with high resolution. In this study, we examined the potential role of CBCT as a tool for 3-dimensional evaluation of the glottal insufficiency. Methods CBCT was performed in 2 normal controls and in 14 patients with unilateral vocal fold paralysis (13 cases) or paresis (1 case). Laryngeal CBCT images were taken in the resting state and during phonation. Furthermore, preoperative and postoperative images were evaluated to assess the impact of phonosurgeries in 5 cases. Results CBCT of the vocalizing vocal fold was successfully performed in all the cases, producing clear coronal images. During phonation, no remarkable differences in levels between the paralyzed and normal vocal folds were observed in 10 cases, proving the 2-dimensional glottal gap (2D cases). On the other hand, the levels of paralyzed vocal folds stayed higher during phonation compared to the normal side in 4 cases, proving the 3-dimensional glottal gap (3D cases). 4 of the 2D cases had injection laryngoplasty and 1 of the 3D cases had arytenoid adduction procedure to correct their glottal gaps. Postoperative CBCT images verified the successful correction of these gaps. Conclusions Our study proved the feasibility of CBCT for 3D evaluation of vocalizing larynx. This quick and easy system may have an informative role to suggest the proper phonosurgical procedure for glottic insufficiency.