Archives of Otol

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American Head & Neck Society
July 21-25, 2012
Metro Toronto Convention Centre
Toronto, ON, Canada

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Presentation: P165
Topic: Basic Science
Type: Poster
Date: Sunday - Tuesday, July 22 - 24, 2012
Session: Designated Poster viewing times
Authors: Shunya Egawa, MD, Shinichi Iwai, MD PhD, Tomohiro Ono, MD, Takehiko Sambe, MD, Takeshi Hayashi, MD, Kakei Ryu 1, DDS, Toshikazu Shimane, MD PhD, Katsuji Oguchi, MD PhD
Institution(s): Department of 1) Pharmacology and 2) Otorhinolaryngology, Showa University School of Medicine. 1-5-8 Hatanoda,Shinagawa-ku,Tokyo,Japan

Introduction: The presence of metastasis is the main prognostic factor of patients with malignant tumors. Cancer cells migration and invasion are important factors for metastasis in oral squamous cell carcinomas (SCCs) such as the tongue and floor of the mouth. The matrix metalloproteases (MMPs) are a family of zinc-dependent proteases involved in the degradation of extracellular matrix components. They are also thought to play a major role on cell behaviors such as cell migration, adhesion/dispersion, invasion and proliferation. Therefore, MMP inhibitor (MMPI) is one of useful agents for treatment of the cancers. Marimastat acts as a broad-spectrum MMPI. We reported previously that (-)-epigallocatechin-3-gallate (EGCG) as a green tea polyphenol had ability of MMPI. Several papers reported EGCG was an anti-cancer agent in vivo and vitro. It was very difficult to observe cancers cells migration and invasion in real time until recently. Recently real-time cell analysis system (RTCA) was developed for cell migration and invasion in vitro. In this study, we investigated that MMPIs agents inhibited the cells migration and invasion in oral SCCs in real time by RTCA. Materials and Methods: We used human SCCs of the tongue cell line SCC-4 and SAS cells, and floor of the mouth cell line H0-1-u-1 cells which were obtained by Health Science Research Resources Bank. Marimastat and EGCG were obtained by Wako. We used the RTCA (Roche) for migration and invasion on SCCs. RTCA was that the more cells were attached on the electrodes in CIM-plate 16, the larger the increased in electrode impedance. Migration was used by CIM-plate 16 with fibronectin coating. SCCs (8.0 or 16.0 x 104 cells/well) were cultured for 24 or 48 hours with CIM-plate 16. Invasion was used by CIM-plate 16 with matrigel coating. The mRNA expression of MMPs was determined by real-time RT-PCR. Results: SCC-4 cells demonstrated high ability for cell migration and invasion compared with SAS and H0-1-u-1 cells by RTCA. The mRNA of MMP-9 in SCC-4 cells increased more than two times compared with SAS and H0-1-u-1 cells. On the other hand, MMP-2 expressions did not have significant changes among SCCs. All examined SCCs were suppressed cell migration and invasion by 100 microM marimastat and EGCG. The cell invasion had inhibition more clear than the cell migration among SCCs by MMPIs. Moreover, The cell invasion of SCC-4 cells was suppressed clearly by MMPIs. As cells with much expression of MMP such as SCC-4, the effect of MMPI is strong. Conclusion: We demonstrated that the cells migration and invasion in oral SCCs were inhibited in real time by RTCA. We were able to capture the individuality of SCCs by RTCA. The cells migration and invasion in oral SCCs were shown that were associated with MMP. Our results suggest that RTCA was useful for cells migration and invasion on oral SCCs in real time. Therefore, RTCA will be useful for development of more effective new MMPI drugs.


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