Cytokeratin 13, Cytokeratin 17, and Ki-67 Expression in Human Acquired Cholesteatoma and Their Correlation With Its Destructive Capacity

OBJECTIVES: Cholesteatoma is a nonneoplastic destructive lesion of the temporal bone with debated pathogenesis and bone resorptive mechanism. Both molecular and cellular events chiefly master its activity. Continued research is necessary to clarify factors related to its aggressiveness. We aimed to investigate the expression of Ki-67, cytokeratin 13 (CK13) and cytokeratin 17 (CK17) in acquired nonrecurrent human cholesteatoma and correlate them with its bone destructive capacity. METHODS: A prospective quantitative immunohistochemical study was carried out using fresh acquired cholesteatoma tissues (n=19), collected during cholesteatoma surgery. Deep meatal skin tissues from the same patients were used as control (n=8). Cholesteatoma patients were divided into 2 groups and compared (invasive and noninvasive) according to a grading score for bone resorption based upon clinical, radiologic and intraoperative findings. To our knowledge, the role of CK17 in cholesteatoma aggressiveness was first investigated in this paper. RESULTS: Both Ki-67 and CK17 were significantly overexpressed in cholesteatoma than control tissues (P < 0.001 for both Ki-67 and CK17). In addition, Ki-67 and CK17 were significantly higher in the invasive group than noninvasive group of cholesteatoma (P=0.029, P=0.033, respectively). Furthermore, Ki-67 and CK17 showed a moderate positive correlation with bone erosion scores (r=0.547, P=0.015 and r=0.588, P=0.008, respectively). In terms of CK13, no significant difference was found between cholesteatoma and skin (P=0.766). CONCLUSION: Both Ki-67 and CK17 were overexpressed in cholesteatoma tissue and positively correlated with bone resorption activity. The concept that Ki-67 can be a predictor for aggressiveness of cholesteatoma was supported. In addition, this is the first study demonstrating CK17 as a favoring marker in the aggressiveness of acquired cholesteatoma.

Immunofluorescence examination of CK-13 expression in cell line KB differentiated by all-trans retinoic acid or As2 O3 / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To examine the expression of cytokeratin-13 (CK-13) in oral squamous cell carcinoma (OSCC) and to discuss the effects of all-trans retinoic acid (ATRA) or arsenic trioxide (As2 O3) on the differentiation of human oral undifferentiated squamous cell carcinoma cell line KB cells.</p><p><b>METHODS</b>The cultured KB cells were divided into three groups, ATRA group, As2 O3 group, and control. The expression of CK-13 in KB cells was detected using the immunofluorescence before and after KB cells were induced by ATRA or As2 O3.</p><p><b>RESULTS</b>The expression rates of CK-13 in KB cells in the ATRA group and As2 O3 group were significantly higher than that in the control (P < 0.05), but there was no significant difference in the expression between ATRA and As2 O3 group(P > 0.05).</p><p><b>CONCLUSIONS</b>ATRA and As2 O3 both have the ability to differentiate the KB cells, and the expression is associated with the degree of tumor differentiation. CK-13 may serve as a molecular marker to evaluate the effect of the differentiation treatment on OSCC.</p>

[ immunohistochemical comparative evaluation of CK13 and CK18 expression in odontogenic cysts and ameloblastoma]

Evaluating cytokeratin contents of odontogenic cysts and tumors can be one of the important aspects in comparative study of these lesions. The aim of this study was to evaluate expression of CK13 and CK18 in odontogenic cysts and ameloblastomas. In this study, radicular cysts [20 cases], dentigerous cysts [20 cases], OKC [20 cases], unicystic ameloblastoma [20 cases] and solid ameloblastoma [20 cases] were evaluated. Then, 3-4 micro m sections were prepared and stained with immuhistochemical procedure [avidin- biotin method. The obtained data by microscopic consideration was analyzed with statistical tests such as: Chi-square, Mann-Whitney, Wilcoxon [p<0.05]. All of the odontogenic cysts and cystic parts of unicystic ameloblastoma expressed CK13 in the upper layers [middle and superficial] more than the deeper ones [basal and parabasal]. In ameloblastomas, CK 13 expression in stellate reticulum and acanthomatose component was more than preameloblasts. Expression of CK 18 in the upper layers [middle and superficial] was more than deeper layers [basal and parabasal] in radicular and dentigerous cysts. Results of immunoreactivity with CK18 in OKC was negative in 100% of cases. In addition, ameloblastomas [unicystic and solid] did have not significant differences in expression of CK13 and CK18. CK13 did not show any differences between these lesions but significant difference in CK18 expression at all 4 layers of the epithelium in odontogenic cysts and ameloblastomas may indicate the different nature of these lesions and probably confirms this suggestion that unicystic ameloblastoma is initiated as a neoplasm essentially, no from neoplastic change of an odontogenic cyst, in particular dentigerous cyst. The same CK 13 and CK18 expression pattern in unicystic and solid ameloblastomas reveals the same biomolecular nature despite different gross and clinical features

Application of protein markers in combination with ThinPrep bronchial brush cytology in classification of lung cancer subtypes / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the value of application of cellular protein markers stained by immunocytochemistry in combination with ThinPrep bronchial brush cytology in classification of lung cancer subtypes.</p><p><b>METHODS</b>Remaining bronchial brush cytology samples from 206 lung cancer patients with positive cytological diagnosis and 45 fine needle aspiration samples of resected lung carcinomas were collected. The expressions of CK10/13, CK7, CK18, CD56 and SYN in those samples were detected by immunocytochemistry (ICC) using corresponding antibodies.</p><p><b>RESULTS</b>The sensitivity and specificity of CK10/13 were 94.7% and 72.0%, respectively, in diagnosis of squamous cell carcinoma. The sensitivity and specificity of CK7 were 98.6% and 61.5%, and those of CK18 were 98.6% and 37.5%, respectively, in diagnosis of adenocarcinoma. The sensitivity and specificity of CD56 were 86.3% and 82.9%, and those of SYN were 81.6% and 93.5%, respectively, in diagnosis of small cell lung cancer. No significant difference was found in the expressions of CK10/13, CK7 and CK18 protein markers among differently differentiated lung squamous cell carcinomas and adenocarcinomas (P > 0.05). The classification rate of cytology in combination with ICC in differential diagnosis for 44 cases of unclassified lung cancer reached 90.0% for squamous cell carcinoma, 96.3% for adenocarcinoma, and 100.0% for small cell lung carcinoma.</p><p><b>CONCLUSION</b>Application of cellular protein markers in combination with ThinPrep bronchial brush cytology is helpful to improve the differential diagnosis of lung cancer subtypes, and may become a supplementary diagnostic method in subclassification of lung cancer.</p>

Cytokeratin18, 13 and their gene expression in post-operative maxillary cyst linings with metaplastic epithelium / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To study the cytokeratin 18 and 13 and their gene (CK) expression in post-operation maxillary cyst linings with metaplastic epithelium.</p><p><b>METHODS</b>CK expressions were examined with immunohistochemistry in 46 post-operative maxillary cyst (POMC) which were lined with pseudostriated columnar cells only (13 cases), both kinds of columnar and squamous cells (30 cases) and squamous cells only (3 cases).</p><p><b>RESULTS</b>The expressions of CK8, CK13 and CK18 were observed in 39, 9 and all of the 43 columnar epithelial linings, respectively. Metaplastic squamous epithelia expressed more CK13 and less CK18 and CK8. Of the 33 metaplastic linings, 24 expressed CK8, 23 CK13 and 26 linings expressed CK18. The expression of CK13- and CK18-mRNA was generally correlated with the protein expression level. By in situ hybridization, CK18-mRNA expression was observed not only in 26 metaplastic linings which were positive for CK18 protein but also in five of the seven metaplastic linings which did not express CK18 protein. In addition, RT-PCR revealed an expression of CK18-mRNA in all metaplastic squamous linings although the expression level was weaker than that in the columnar epithelial linings. The CK13-mRNA was expressed in a fashion inverse to the CK18-mRNA.</p><p><b>CONCLUSIONS</b>These results indicate that CK18-mRNA is preserved through metaplasia although the protein expression decreases and metaplastic squamous cells differentiate with a decrease of CK18 and an increase of CK13 expression.</p>

Three-dimensional Culture Model of the Conjunctival Epithelium

PURPOSE: To reconstruct a cultured conjunctival equivalent that closely resembles normal conjunctival epithelium in three-dimensional culture systems. METHODS: Human conjunctival epithelial cells were cultured on dead de-epidermized dermis in the air-exposed state. After 2 weeks of culture, the sections were stained with hematoxylin and eosin. Immunohistochemical and electron microscopic studies were performed. The results were compared with those of normal conjunctiva and cultured eyelid skin equivalent. RESULTS: In the cultured conjunctival equivalent, nonkeratinizing stratified epithelium was formed similarly to normal conjunctival epithelium. Keratin 13 was expressed, but not keratin 10, in the cultured conjunctival equivalent, similarly to normal conjunctival epithelium. However, in the cultured eyelid skin equivalent, keratinizing stratified epithelium was formed. In addition, keratin 10 was expressed, but not keratin 13, contrary to those of the cultured conjunctival equivalent. In the cultured conjunctival equivalent, ultrastructurally, keratin intermediate filaments and desmosomes were found. In addition, microvilli were seen in the uppermost epithelial cells. CONCLUSIONS: These findings demonstrate that the cultured conjunctival equivalent resembles normal conjunctival epithelium morphologically, biochemically and ultrastructurally, thereby suggesting that the cultured conjunctival equivalent may have a great potential in the study of conjunctival epithelium.

Proteomic analyses of cervical cancer tissues by two-dimensional gel electrophoresis and mass spectrometry / 대한산부인과학회지

OBJECTIVE: Comparison of protein expressions by two-dimensional gel electrophoresis (2-DE) in normal cervix and squamous cell carcinoma tissues in Korean women. METHODS: Normal cervix and squamous cell carcinoma tissues were solubilized with 2-DE buffer and the first dimension of PROTEAN IEF CELL, isoelectric focusing (IEF), was performed using pH3-10 linear IPG strips of 17 cm. And then running 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and sliver stain. Scanned image was analyzed using PDQuest 2-D softwareTM. Protein spot spectrum was identified by assisted laser desorption/ionization-time of fighting (MALDI-TOF) and the protein mass spectrum identifications were performed by searching protein databases of Swiss-prot/TrEMBL, Mascot and MS-FIT. RESULTS: We found 9 up-regulation proteins (Alpha enolase, Keratin 19 type I, Keratin 20 type I, Keratin 13 type I, beta-actin, Aflatoxin B1 aldehyde reductase 1, Annexin A2, Squamous cell carcinoma antigen 2, unknown), 7 down-reguation proteins (Annexin 1, Myosin regulatory light chain 2, 14-3-3 protein epsilon, Heat shock 27 kDa protein, Hypothetical protein (DKFZP434C1715), Tumor necrosis factor receptor superfamily member 13B, Smoth muscle protein 22-alpha) and 6 up and down-regulation proteins (Tropomyosin 1, Tropomyosin 2, Tropomyosin 3, Serine (or cysteine) proteinase inhibitor, Phosphatidylinositol transfer protein alpha isoform, Src homology 3 domain-containing protein HIP-55) between normal cervix and squamous cell carcinoma cell tissues. CONCLUSION: 2-DE offers total protein expressions between normal cervix and squamous cell carcinoma cell tissues, and searching of differently expressed protein for the diagnostic markers of squamous cell carcinoma tissue.

A Case of Squamous Cell Carcinoma of Thymus with Negative CD5, Cytokeratin 7 and Positive Cytokeratin 13 / 결핵

The most common anterior mediastinal tumors originate from the thymus. Among them, thymic carcinomas occur as an early local invasion and wide spread metastases. However, when squamous cell carcinoma in the thymus or mediastinum is identified, an occult primary lung cancer must be excluded because the histologic types resemble those found more typically in the lung. CD5 and cytokeratin immunohistochemical staining is useful in evaluating biopsy samples from those tumors. Spuamous cell carcinoma of an unknown primary origin in the mediastinum is a rare occurrence and there are only a handful of case reports. Here we describe a case with an anterior mediastinal mass of squamous cell carcinoma with unknown primary origin. A resection of the mediastinal mass without an association with the lung was performed. Immunohistochemical stainings were positive using cytokeratin 13, and negative using CD5 and cytokeratin 7. This was followed by chemotherapy for presured thymic carcinoma.

Expression of Epidermal Protein Antigens and Basement Membrane Components in Human Epidermis Reconstructed by Culture / 대한피부과학회지

BACKGROUND: A number of in vitro skin models have been developed for the purpose of the screening of cosmetics, pharmaceuticals, and environmental chemicals. To mimic the skin in vivo, a model should resemble morphologically and biochemically the parent, tissue. OBJECTIVE: The purpos of this study is to study the differentiation and organization of the artificial epidermis in comparsion with epidermis in vivo based on the expression of epidermal protein antigens and basement membrane components. METHODS: Human keratinocytes were cultured on deepidermidized dermis (RE-DED) or on fibroblast-populated collag-,n matrix (LSE). After 10 days culture, the sections of RE-DED and LSE were stained with hematoxylin and eosin. An immunohistochemical study was also performed with the sections of RE-DED and LSE using antibodies recognizing proliferating cell nuclear antigens (PCNA), epidermal growth factor receptor (EGFR), keratin 1, involucrin, filaggrin, loricrin, keratin 13, type IV collagen, and laminin. RESULTS: In both culture systems(RE-DED and LSE) a multilayered epidermis with a horny layer was observed. In the human epidermis reconstructed by both culture systems, differentiation markers appeared but with a topography slightly different from that of epidermis in vivo, and components of the basement membrane was also expressed. CONCLUSION: Our findings suggest the epidermis obtained in both culture systems(RE-DED and LSE) resembled in vivo epidermis morphologically and biochemically, although it was not the same.

Expression of Epidermal Protein Antigens and Basement Membrane Components in Human Epidermis Reconstructed by Culture / 대한피부과학회지

BACKGROUND: A number of in vitro skin models have been developed for the purpose of the screening of cosmetics, pharmaceuticals, and environmental chemicals. To mimic the skin in vivo, a model should resemble morphologically and biochemically the parent, tissue. OBJECTIVE: The purpos of this study is to study the differentiation and organization of the artificial epidermis in comparsion with epidermis in vivo based on the expression of epidermal protein antigens and basement membrane components. METHODS: Human keratinocytes were cultured on deepidermidized dermis (RE-DED) or on fibroblast-populated collag-,n matrix (LSE). After 10 days culture, the sections of RE-DED and LSE were stained with hematoxylin and eosin. An immunohistochemical study was also performed with the sections of RE-DED and LSE using antibodies recognizing proliferating cell nuclear antigens (PCNA), epidermal growth factor receptor (EGFR), keratin 1, involucrin, filaggrin, loricrin, keratin 13, type IV collagen, and laminin. RESULTS: In both culture systems(RE-DED and LSE) a multilayered epidermis with a horny layer was observed. In the human epidermis reconstructed by both culture systems, differentiation markers appeared but with a topography slightly different from that of epidermis in vivo, and components of the basement membrane was also expressed. CONCLUSION: Our findings suggest the epidermis obtained in both culture systems(RE-DED and LSE) resembled in vivo epidermis morphologically and biochemically, although it was not the same.