{"created":"2023-06-23T12:22:20.801480+00:00","id":14195,"links":{},"metadata":{"_buckets":{"deposit":"924d20c6-c94a-487e-bc3f-8a0bf07adb69"},"_deposit":{"created_by":21,"id":"14195","owners":[21],"pid":{"revision_id":0,"type":"depid","value":"14195"},"status":"published"},"_oai":{"id":"oai:asahi-u.repo.nii.ac.jp:00014195","sets":["46:348:501"]},"author_link":["3561","16014","1290","16934","16620","4301","4042","16581","4109","8868","2875","3998","4090","14482","496","16580"],"item_10002_biblio_info_77":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2022-10","bibliographicIssueDateType":"Issued"},"bibliographicIssueNumber":"2","bibliographicPageEnd":"80","bibliographicPageStart":"73","bibliographicVolumeNumber":"49","bibliographic_titles":[{"bibliographic_title":"岐阜歯科学会雑誌"},{"bibliographic_title":"The Journal of Gifu Dental Society","bibliographic_titleLang":"en"}]}]},"item_10002_description_80":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"口腔粘膜細胞診は、非侵襲性に実施できる利点がありPapanicolaou染色（PAP 染色）による細胞や核の形態的変化を細胞異型として判定している。しかし、その判定には熟練した専門的知識を必要とし、これを裏付ける客観的判定基準の統一が求められている。そこで本研究では、口腔粘膜細胞診検体のPAP染色でみられる形態的変化を細胞骨格関連分子の変化で捉えて、客観的判定基準に裏付けるために検索を行った。実験は診断後の液状化検体細胞診（Liquid based cytology, LBC）標本と10％ホルマリンで固定したLBC（FLBC）標本を用いてCK（AE1/AE3）、F-actin、Cortactinに対する蛍光染色とDAPIによる核蛍光染色を行い、PAP染色との比較を行った。また組織標本にて扁平上皮癌及び健全粘膜組織と確認できた症例を用いてCortactinに対する免疫染色を行い、光学顕微鏡で観察した。1．CK染色に対する免疫蛍光染色において、細胞質や核の蛍光強度は，腫瘍性変化に伴って上昇し、PAP染色の細胞質や核の変化にそれぞれ対応していた。2．F-actin染色では、NILMとSCCでは微細構造に変化を認め、悪性化に伴って糸状仮足や細胞外に突出した結節状変化を認めた。なお、共焦点レーザー顕微鏡の画像解析ソフトから算出したN/C比で有意差検定をしたところ、NILMと比較してSCCで有意な上昇を示した。3．F-actinとCortactinの二重染色における共発現は、異型を示す深層系細胞の浸潤突起にのみ限局して発現を認めた。組織標本から検索したCortactinの発現は、正常組織では有棘層および基底層に弱陽性像を認め、SCC周辺の上皮性異形成部では、中間層から基底層にかけて陽性像を認めた。SCCでは10症例中7例で深部および浸潤癌胞巣の最外層細胞に強い陽性像を認めた。以上の結果から、LBC法を用いた口腔粘膜細胞診と共焦点レーザー顕微鏡による細胞骨格関連分子や核の蛍光標識は、PAP染色の形態的所見の裏付けとなるだけでなく、細胞診判定の客観的指標を持った根拠として、腫瘍性変化の早期発見・早期治療や細胞診のAI化に繋げることができると考えた。","subitem_description_type":"Abstract"}]},"item_10002_description_81":{"attribute_name":"内容記述","attribute_value_mlt":[{"subitem_description":"In the oral region, exfoliative cytology is a relatively non-invasive procedure. Specimens from oral mucosa are stained by Papanicolaou (PAP) technique to identify cellular atypia. The present criteria to diagnose such orphological changes is subjective; therefore, objective criteria should be established on the basis of histological evidence. We investigated the morphological changes of oral epithelial cells with PAP and florescent stainings, focusing on changes in cytoskeleton-related molecules. Liquid-based cytology (LBC) and LBC fixed with 10% formalin were used for exfoliative specimens, which were previously diagnosed as negative for intraepithelial lesion or malignancy (NILM), low-grade or high-grade dysplasia or squamous cell carcinoma (SCC). Also we used 10% formalin-fixed paraffin sections of tongue SCC including the dysplastic area around carcinoma and normal buccal mucosa. The cytoskeleton-related molecules including pan-cytokeratin (CK) AE1/AE3, F-actin and cortactin were labeled fluorescently, and nuclei were labeled with 4’5-diamidino-2-phenylindole dihydrochloride. We observed these molecules and nuclei by using confocal laser scanning microscopy, and examined the findings of these fluorescent and PAP stainings. We also performed immunohistochemistry for cortactin in the specimens of the tongue SCC and the normal buccal mucosa. In immunofluorescence staining of CK AE1/AE3, immunofluorescence intensities in cytoplasm and nuclei were increased as the neoplastic change progressed, which corresponded to the changes in intensities observed in PAP-stained cytoplasm and nuclei. With F-actin specific fluorescent phalloidin, changes in microstructures of cells were observed in SCC and NILM epithelial cells, showing filamentous pseudopodia or extracellular nodular form associated with cell malignancy. The nucleus-to-cytoplasm ratio was calculated by using the image analysis, and the ratio of SCC cells was significantly higher than that of NILM cells. In the double staining of F-actin and cortactin, the co-expression of these two molecules was localized in the invadopodia of the deeper tumor cells. Strong immunohistochemical expression of cortactin was observed in 7 of 10 SCC sections, and their intensities were stronger in the deeper region and outermost layer of invasive carcinoma nests than in the dysplastic area around carcinoma, whereas weak expression of cortactin was observed in the stratum spinosum and the basal layers of normal tissues. In conclusion, the use of LBC and microscopic observation of fluorescent-labeled cytoskeleton-related molecules and nuclei can support the morphological findings observed in PAP-stained smears. Therefore, our results may contribute to applying objective criteria for cytodiagnosis and the development of deep learning artificial intelligence technology that will facilitate the early detection of oral cancer. 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Dentistry"},{"subitem_textarea_value":"朝日大学歯学部口腔病態医療学講座口腔外科学分野\nDepartment of Oral & Maxillofacial Surgery Division of Oral Pathogenesis & Disease Control Asahi University School of Dentistry"},{"subitem_textarea_value":"明海大学保健医療学部\n順天堂大学医学部附属浦安病院口腔ケア室\nSchool of Health Sciences, Meikai University\nDepartment of Oral Care, Juntendo Urayasu Hospital"}]},"item_10002_version_type_95":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_970fb48d4fbd8a85","subitem_version_type":"VoR"}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorAffiliations":[{"affiliationNameIdentifiers":[{"affiliationNameIdentifier":"33703","affiliationNameIdentifierScheme":"kakenhi","affiliationNameIdentifierURI":"https://kaken.nii.ac.jp/ja/search/?qm=33703"}],"affiliationNames":[{"affiliationName":"朝日大学","affiliationNameLang":"ja"}]}],"creatorNames":[{"creatorName":"篠島, 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kB"}],"format":"application/pdf","licensetype":"license_11","mimetype":"application/pdf","url":{"label":"gifushika492_7380_2022","url":"https://asahi-u.repo.nii.ac.jp/record/14195/files/gifushika492_7380_2022.pdf"},"version_id":"819599dd-c5b7-43d5-ab3a-b74ab996c55a"}]},"item_keyword":{"attribute_name":"キーワード","attribute_value_mlt":[{"subitem_subject":"共焦点レーザー顕微鏡","subitem_subject_scheme":"Other"},{"subitem_subject":"液状化検体細胞診","subitem_subject_scheme":"Other"},{"subitem_subject":"Cytokeratin","subitem_subject_scheme":"Other"},{"subitem_subject":"F-actin","subitem_subject_scheme":"Other"},{"subitem_subject":"Cortactin","subitem_subject_scheme":"Other"},{"subitem_subject":"Confocal laser scanning microscopy","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Liquid based cytology","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Cytokeratin","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"F-actin","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Cortactin","subitem_subject_language":"en","subitem_subject_scheme":"Other"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"jpn"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"departmental bulletin paper","resourceuri":"http://purl.org/coar/resource_type/c_6501"}]},"item_title":"口腔粘膜細胞診で判定する上皮細胞の形態学的特徴を裏付ける細胞骨格分子","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"口腔粘膜細胞診で判定する上皮細胞の形態学的特徴を裏付ける細胞骨格分子"},{"subitem_title":"Cytoskeletal molecules supporting the morphological characteristics of epithelial cells determined by oral mucosal cytology","subitem_title_language":"en"}]},"item_type_id":"10002","owner":"21","path":["501"],"pubdate":{"attribute_name":"公開日","attribute_value":"2023-04-25"},"publish_date":"2023-04-25","publish_status":"0","recid":"14195","relation_version_is_last":true,"title":["口腔粘膜細胞診で判定する上皮細胞の形態学的特徴を裏付ける細胞骨格分子"],"weko_creator_id":"21","weko_shared_id":-1},"updated":"2024-02-17T02:49:06.353947+00:00"}