OBJECTIVETo study the cytokeratin 18 and 13 and their gene (CK) expression in post-operation maxillary cyst linings with metaplastic epithelium.

METHODSCK 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).

RESULTSThe 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.

CONCLUSIONSThese 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.

Epithelial Cells ; metabolism ; pathology ; Humans ; Jaw Cysts ; etiology ; metabolism ; pathology ; Keratin-13 ; biosynthesis ; genetics ; Keratin-18 ; biosynthesis ; genetics ; Maxillary Diseases ; etiology ; metabolism ; pathology ; Metaplasia ; metabolism ; pathology ; Postoperative Complications ; RNA, Messenger ; genetics

UNLABELLED: Abstract OBJECTIVE: To measure the expression of CK18 and CK19 in the cells from peripheral blood and tumor tissue of the nasopharyngeal carcinoma patients,to test whether CK 18 and CK 19 could be biomarkers of nasopharyngeal carcinoma fordiagnosis. METHOD: The mRNA was extracted from the blood and carcinoma tissue of nasopharyngeal carcinoma and was reversed transcription to cDNA. The 3 pairs primers were designed for RT-PCR and the fold value was calculated to evaluated expression by ΔCT. RESULT: There are no statistical differences between the CK18 and CK19 gene expression and the gender, age and metastasis in tumor tissue of 45 nasopharyngeal carcinoma patients (P>0. 05). There are significant differences among 3 pathological stages and 2 genes expressed increase as the grade malignancy (P<0. 05). The detecting of the 2 genes expression from blood cells shows that CK18 and CK19 had a high positive ratio 64% and 75% respectively. Meanwhile this method showed a same detection characteristic in tumor and blood, the positive.rate of CK18 and CK19 genes in metastasis is higher than non-metastasis. The results showed CK18 has a high specificity and CK19 has a high sensitivity for prognosis and all relapsed cases are associated with the expression of CK18 and CK19. CONCLUSION CK18 and CK19 may be used as biomarkers of nasopharyngeal carcinoma for diagnosis.
Biomarkers, Tumor ; Carcinoma ; DNA, Complementary ; Gene Expression ; Humans ; Keratin-18 ; biosynthesis ; Keratin-19 ; biosynthesis ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; diagnosis ; metabolism ; pathology ; Neoplasm Metastasis ; Prognosis ; RNA, Messenger ; Reverse Transcriptase Polymerase Chain Reaction

Pleomorphic carcinoma of the lung (PCL) is characterized by a mixture of sarcomatoid and carcinoma components, and a poor prognosis. However, no immunophenotype of tumor markers has been characterized in PCL. To charaterize the immunophenotype for CD99 in PCL, we performed an immunohistochemical evaluation of PCLs for thyroid transcription factor-1 (TTF-1), cytokeratin (CK) 7 and 20, and for CD99. CD99 was found to be expressed in both carcinomatous (47%) and sarcomatous components such as spindle cells (92%) and giant cells (57%). In the case of spindle cells, CK7 was expressed in 6 cases (46%) and TTF-1 in 2 cases (15%), whereas for giant cells CK7 was expressed in 8 cases (57%) and TTF-1 in one case (7%). However, CK20 was not expressed in either the carcinomatous or sarcomatous components in any case. Thus, CD99 was found to be widely expressed in both sarcomatous and carcinoma component in PCL. A clinicopathological analysis showed no direct correlation between the expression of CD99 and the clinical indices (stage, survival rate, invasion) of PCL.
Adult ; Aged ; Aged, 80 and over ; Antigens, CD/*biosynthesis ; Carcinoma/*metabolism ; Cell Adhesion Molecules/*biosynthesis ; Female ; Humans ; Immunohistochemistry ; Immunophenotyping ; Intermediate Filament Proteins/biosynthesis ; Keratin/biosynthesis ; Lung Neoplasms/*metabolism ; Male ; Middle Aged ; Nuclear Proteins/biosynthesis ; Prognosis ; Research Support, Non-U.S. Gov't ; Sarcoma/metabolism ; Time Factors ; Transcription Factors/biosynthesis

OBJECTIVETo detect the expression of cytokeratin 20 (CK20) mRNA in peripheral blood of colorectal carcinoma and to discuss its clinical value.

METHODSReal-time fluorescent quantitative RT-PCR was used to detect the CK20 mRNA expression in the peripheral blood of 51 patients with colorectal carcinoma and 30 healthy volunteers.

RESULTS27.45% of the patients showed CK20 mRNA expression, while it was 6.67% for the control group (P<0.025). With the progress of Dukes' stages, the expression level of CK20 mRNA increased, but there was no statistic significance (P<0.05). More samples in Dukes'C and D than in Dukes'A and B stages showed >10 copies/ml.

CONCLUSIONThe detection of CK20 mRNA expression in peripheral blood of patients with colorectal carcinoma may be helpful to identify early shedding tumor cells. It is also useful to monitor the progression of the disease and observe the effect of clinical treatment.

Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; blood ; genetics ; metabolism ; Female ; Humans ; Intermediate Filament Proteins ; biosynthesis ; blood ; genetics ; Keratin-20 ; Lymphocytes ; metabolism ; Male ; Middle Aged ; RNA, Messenger ; biosynthesis ; blood ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods

Recently, the rearrangement of RET proto-oncogene has been reported to be the most common genetic change in papillary thyroid carcinoma (PTC). However, its prevalence has been reported variably and its relation to clinical outcome has been controversial. The characteristic nuclear features of PTC usually render the diagnosis, but problem arises with equivocal cytologic features that are present focally. Although there remains some controversy, CK19 has been reported to be a useful ancillary tool for diagnosis of PTC. To evaluate the expression rate of RET/PTC rearrangement and CK19 in PTCs in a Korean population, we studied 115 papillary thyroid carcinomas in 3 mm-core tissue microarray based immunohistochemical analysis. The prevalence of Ret protein expression was 62.6% and the CK19 immunoreactivity was 80.9%. There was no statistically significant asso-ciation between the Ret positivity and CK19 immunoreactivity, although the percent agreement of the two was relatively high. The clinicopathological variables did not correlate with the expression of Ret. In conclusion, the prevalence of Ret protein expression and its clinicopathological implications in a Korean population are not much different from those reported in previous studies. However, its detection via immunohistochemistry can be a useful diagnostic tool for diagnosing papillary thyroid carcinoma in conjunction with CK19.
Adenocarcinoma, Papillary/*metabolism ; Adult ; Carcinoma/pathology ; Cell Line, Tumor ; Cytoplasm/metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Keratin/*biosynthesis ; Korea ; Lymphatic Metastasis ; Male ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; Oncogene Proteins/*biosynthesis ; Receptor Protein-Tyrosine Kinases/*biosynthesis ; Thyroid Neoplasms/*metabolism/pathology

This study is aimed at establishing a sensitive approach to detect disseminated tumor cells in peripheral blood and evaluate its clinical significance. A total of 198 blood samples including 168 from colorectal carcinoma (CRC) patients and 30 from healthy volunteers were examined by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) to evaluate the expression of carcinoembryonic antigen (CEA), cytokeratin 20 (CK20) and cytokeratin 19 (CK19) mRNA. CEA mRNA was detected in 35.8% of patients and 3.3% of controls, CK20 mRNA in 28.3% of patients and 6.7% of controls, and CK19 mRNA in 41.9% of patients and 3.3% of controls. CEA and CK20 mRNA positive ratio increased with the advancing Dukes stages, but there was no significant difference in positive ratio between any two stages (P>0.05). Also, relatively high positive ratio of CEA, CK20 and CK19 mRNA expression was observed in some CRC patients with earlier Dukes stages. A higher positive ratio was obtained when two or three detection markers were combined compared to a single marker. Our study indicates that quantitative real-time RT-PCR detection for CEA, CK20 and CK19 mRNA in peripheral blood is a valuable tool for monitoring early stage dissemination of CRC cells in blood circulation.
Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; biosynthesis ; blood ; genetics ; Carcinoembryonic Antigen ; biosynthesis ; blood ; genetics ; Carcinoma ; blood ; genetics ; Colorectal Neoplasms ; blood ; genetics ; Female ; Humans ; Keratin-20 ; Keratins ; biosynthesis ; blood ; genetics ; Male ; Middle Aged ; RNA, Messenger ; blood ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity

The purpose of this study is to characterize and compare the ultrastructural changes occurring during the in vivo cultivation of corneal epithelium on amniotic membrane (AM) at several different time points. Corneal burn patients (n=7) with a corneal epithelial defect and severe limbal damage were selected. Initially, AM transplantation with limbal autograft was performed at the acute stage of corneal burn to reconstruct the damaged ocular surface. One to six (mean interval; 3.3+/-1.2) months later, the central part of AM containing an in vivo expanded corneal epithelium was excised and retransplanted in adjacent lesions. The excised epithelium with AM was examined by electron microscopy and immunohistochemical study. By electron microscopy, one and two months after expansion, cultivated epithelium on AM showed an undifferentiated epithelium and an incomplete basement membrane (BM). But, after three months, the cultivated epithelium began to differentiate into a multilayered epithelium with a continuous BM with increased hemidesmosomes. These findings were further confirmed by immunohistochemical study, that cytokeratin K3 was expressed in the cultivated corneal epithelium and newly formed BM was partially positive of collagen IV at three months. At least 3 months may be needed for the proliferation and differentiation of in vivo cultivated corneal epithelium on AM.
Stem Cells/cytology ; Stem Cell Transplantation/*methods ; Middle Aged ; Microscopy, Electron ; Male ; Keratin-3/biosynthesis ; Immunohistochemistry ; Humans ; Epithelium, Corneal/cytology/*metabolism/*pathology/*transplantation ; Corneal Diseases/*therapy ; Burns/*surgery/therapy ; Biological Dressings ; Amnion/*ultrastructure ; Adult

OBJECTIVETo elucidate the significance of detecting cytokeratin gene (CK-20 mRNA) expression in metastatic lymph nodes of colon carcinoma patients.

METHODSThe expression of CK-20 mRNA was detected in 342 lymph nodes by reverse transcription-polymerase chain reaction (RT-PCR) in 49 colon carcinoma patients. Its incidence was compared with that by routine pathologic examination.

RESULTSThe positive rates of detecting metastasis in the lymph nodes were 21.9% by RT-PCR and 11.1% by routine pathologic examination.

CONCLUSIONThe detection of cancer metastasis in the lymph nodes in colon carcinoma is almost doubled (21.9% vs 11.1%) by CK-20 mRNA, which may provide a guiding significance in staging, treatment planning and prognosis in the prognostic estimation of colon carcinoma patients.

Adult ; Aged ; Colonic Neoplasms ; diagnosis ; genetics ; pathology ; Female ; Gene Expression ; Genetic Markers ; genetics ; Humans ; Intermediate Filament Proteins ; genetics ; Keratin-20 ; Lymphatic Metastasis ; diagnosis ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; RNA, Messenger ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the differentiation of bone marrow derived Thy-1(+)beta(2)M(-) cells (BDTC) into mature and functional liver cells and its mechanism.

METHODSBDTC were cocultured with allyl alcohol (AA)-injured hepatocytes and cultured alone in conditional medium containing HGF and bFGF, respectively. BDTC morphologic transformation was observed with phase-contrast and electron-microscopy. Hepatocyte-specific gene expression in cultured BDTC was identified by immunocytochemistry and reverse transcription-polymerase chain reaction (RT-PCR). Indocyanine green (ICG) ingestion/excretion and urea, albumin production were carried out to evaluate hepatocyte-related function.

RESULTSSome BDTC derived hepatocyte-like cells with high nuclear to cytoplasmic ratio containing mono- or multi-nuclei and abundant mitochondria, endoplasmic reticulum and glycogenic granules appeared after 7-day culture in both the two culture systems. These cells expressed hepatocyte-specific genes (AFP, OV-6, CK18, etc.), and possessed functions of ICG uptake, albumin production and ammonium metabolism.

CONCLUSIONRat BDTCs could differentiate into mature and functional liver cells in special stimulation systems. Moreover, these differentiations were realized by "transdifferentiation", and might dispense with "cell fusion".

Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; Cells, Cultured ; Coculture Techniques ; Fibroblast Growth Factor 2 ; pharmacology ; Gene Expression ; drug effects ; Hepatocyte Growth Factor ; pharmacology ; Hepatocytes ; cytology ; drug effects ; metabolism ; Immunohistochemistry ; Immunomagnetic Separation ; Keratin-18 ; biosynthesis ; genetics ; Propanols ; pharmacology ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Thy-1 Antigens ; metabolism ; alpha-Fetoproteins ; biosynthesis ; genetics

OBJECTIVETo induce the differentiation of human bone marrow mesenchymal stem cells (HMSCs) into hepatocyte-like cells with hepatocyte growth factor (HGF) and fibroblast growth factor-4 (FGF-4) in vitro.

METHODSHMSCs were induced to differentiate into hepatocyte-like cells by HGF (group B), FGF-4 (group C) and HGF+FGF-4 (group D) in vitro. Undifferentiated HMSCs and L-02 cells were used as the negative (group A) and positive (group E) controls, respectively. The changes of cell morphology were observed microscopically. The expressions of hepatic markers, alpha fetoprotein (AFP) and CK-18, were detected by immunocytochemical staining at different times after induction, and the differentiation ratios of the various groups of HMSCs were calculated on the basis of image analysis. The expressions of AFP and ALB were detected by immunofluorescence assay in each group at different times after induction, and the expressions of AFP and ALB mRNA by RT-PCR.

RESULTSHMSCs gradually transformed into spindle-shaped, round, polygonal or irregular cells after induction. Immunocytochemical staining revealed positive AFP and CK18 expressions in groups B, C, and D after induction as well as in group E. The positive units (PU) of AFP and CK18 in group D calculated according to image analysis were significantly higher than that of groups A, B, and C. The expressions of AFP and ALB detected by immunofluorescence were both positive after induction in all groups except group A, similar to the findings of the expressions of AFP and ALB mRNA by RT-PCR.

CONCLUSIONHMSCs can be induced to differentiate into hepatocyte-like cells by HGF, FGF-4 and their combination at certain concentrations, and the hepatocyte-like cells can express some hepatic markers such as AFP, ALB, CK18, etc. HGF+FGF-4 may achieve more effective induction of HMSC differentiation into hepatocyte-like cells, and the efficiency of HGF is greater than that of FGF-4.

Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Cell Differentiation ; drug effects ; Cells, Cultured ; Fibroblast Growth Factor 4 ; pharmacology ; Hepatocyte Growth Factor ; pharmacology ; Hepatocytes ; cytology ; drug effects ; metabolism ; Humans ; Immunohistochemistry ; Keratin-18 ; biosynthesis ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; alpha-Fetoproteins ; biosynthesis ; genetics