Fukuoka Hideki, Kawasaki Satoshi, Yokoi Norihiko, Yamasaki Kenta, Kinoshita Shigeru
Division of Ophthalmology, Department of Advanced Medicine, National Center for Geriatrics and Gerontology, Obu, Japan; Shiley Eye Institute, University of California, San Diego, Calif., USA.
Department of Ophthalmology, Osaka University Graduate School of Medicine, Osaka, Japan; Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Case Rep Ophthalmol. 2016 May 12;7(1):253-61. doi: 10.1159/000445937. eCollection 2016 Jan-Apr.
To report the cytopathological features of corneal intraepithelial neoplasia (CIN) through the investigation of cytokeratin expression pattern, keratinization, cell proliferation, apoptosis, and epithelial mesenchymal transition.
Corneal tissue excised from a CIN patient was examined in this study. Cryosections of the excised CIN epithelial tissue were examined by immunostaining analysis using antibodies against cytokeratins, keratinization-related proteins, Ki-67, human telomerase reverse transcriptase (hTERT), and epithelial mesenchymal transition (EMT)-related proteins. Subcellular localization of F-actin was also analyzed using phalloidin. For the detection of apoptotic cells, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was performed. Real-time polymerase chain reaction was performed to quantify the expression level of hTERT in the CIN epithelium.
The CIN epithelium exhibited a significantly altered cytokeratin expression pattern compared to normal corneas with an upregulated expression of keratinization-related proteins. The CIN epithelium also demonstrated an increased number of Ki-67-positive cells with an upregulated expression of hTERT, while exhibiting an increased number of apoptotic cells. EMT did not occur in the CIN epithelium.
CIN epithelium seems to be slightly dedifferentiated from the corneal epithelial lineage. The status of cell proliferation and apoptosis in the CIN epithelium was significantly altered from that of normal corneal epithelium, but its malignancy level does not appear to be as high as that of metastasis-competent malignant cancers.
通过研究细胞角蛋白表达模式、角化、细胞增殖、凋亡及上皮-间质转化，报告角膜上皮内瘤变（CIN）的细胞病理学特征。
本研究检查了从一名CIN患者切除的角膜组织。使用针对细胞角蛋白、角化相关蛋白、Ki-67、人端粒酶逆转录酶（hTERT）及上皮-间质转化（EMT）相关蛋白的抗体，通过免疫染色分析对切除的CIN上皮组织冰冻切片进行检查。还使用鬼笔环肽分析F-肌动蛋白的亚细胞定位。为检测凋亡细胞，进行了末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）检测。进行实时聚合酶链反应以定量CIN上皮中hTERT的表达水平。
与正常角膜相比，CIN上皮表现出明显改变的细胞角蛋白表达模式，角化相关蛋白表达上调。CIN上皮还显示Ki-67阳性细胞数量增加，hTERT表达上调，同时凋亡细胞数量增加。CIN上皮未发生EMT。
CIN上皮似乎从角膜上皮谱系略有去分化。CIN上皮中细胞增殖和凋亡状态与正常角膜上皮相比有显著改变，但其恶性程度似乎不如有转移能力的恶性肿瘤高。
