12-O-Tetradecanoylphorbol-13-Acetate Induces Keratin 8 Phosphorylation and Reorganization via Expression of Transglutaminase-2

The stiffness of cancer cells is attributable to intermediate filaments such as keratin. Perinuclear reorganization via phosphorylation of specific serine residue in keratin is implicated in the deformability of metastatic cancer cells including the human pancreatic carcinoma cell line (PANC-1). 12-O-Tetradecanoylphorbol-13-acetate (TPA) is a potent tumor promoter and protein kinase C (PKC) activator. However, its effects on phosphorylation and reorganization of keratin 8 (K8) are not well known. Therefore, we examined the underlying mechanism and effect of TPA on K8 phosphorylation and reorganization. TPA induced phosphorylation and reorganization of K8 and transglutaminase-2 (Tgase-2) expression in a time- and dose-dependent manner in PANC-1 cells. These effects peaked after 45 min and 100 nM of TPA treatment. We next investigated, using cystamine (CTM), Tgase inhibitor, and Tgase-2 gene silencing, Tgase-2's possible involvement in TPA-induced K8 phosphorylation and reorganization. We found that Tgase-2 gene silencing inhibited K8 phosphorylation and reorganization in PANC-1 cells. Tgase-2 gene silencing, we additionally discovered, suppressed TPA-induced migration of PANC-1 cells and Tgase-2 overexpression induced migration of PANC-1 cells. Overall, these results suggested that TPA induced K8 phosphorylation and reorganization via Tgase-2 expression in PANC-1 cells.

Construction of a GFP/Puro double-labeled lentiviral vector containing CK8 interfering RNA and its effect on cell apoptosis in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a GFP/Puro double-labeled lentiviral expression vector for CK8 silencing and assess the effects of CK8 silencing on cell apoptosis.</p><p><b>METHODS</b>The siRNA sequences of CK8 were inserted into the lentiviral expression vector GV248 and transfected into 293T cells with the packaging plasmids PMD and SPA. The lentivirus was collected at 24 and 36 h post-transfection. Flow cytometry was used to detect the virus titer and the positive cells were selected with puromycin. The knockdown of CK8 was examined by Western blotting. The effect of CK8 down-regulation on cell apoptosis induced by cisplatin was detected with Annexin V/PI staining.</p><p><b>RESULTS AND CONCLUSION</b>We successfully constructed CK8 interference lentiviral vector and obtained a stable cell line with CK8 knock-down that was sensitive to cisplatin-induced apoptosis.</p>

Morphology, immunohistochemistry and hTERC gene in-situ hybridization in Barrett's esophagus / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinicopathologic features and differential diagnosis of proximal gastric mucosa and mucosa of Barrett's esophagus (BE) in biopsy specimens.</p><p><b>METHOD</b>Thirty-eight cases of Barrett's esophagus (diagnosed using WHO criteria) and 44 cases of proximal gastric mucosa were studied by immunohistochemistry (for CK7, CK20, CK4, CK8, S-100 protein, MUC6, COX2 and bcl-2) and fluorescence in-situ hybridization (FISH) (for hTERC gene). The pathologic features were analyzed.</p><p><b>RESULTS</b>The differences in expression of CK7, CK20, MUC6, COX2 and bcl-2 between BE and proximal gastric mucosa with intestinal metaplasia were not statistically significant (P > 0.05). There was however a statistically significant difference in expression of S-100 protein (P < 0.05). The expression of CK7/CK4 and CK7/CK8 in BE showed positive correlation (P < 0.05). However, such correlation was not demonstrated in proximal gastric mucosa (P > 0.05). The results of hTERC gene expression by FISH showed a statistically significant difference between the two groups: 57.9% (22/38) in BE and 13.6% (6/44) in proximal gastric mucosa (P < 0.05).</p><p><b>CONCLUSIONS</b>The significance of CK7 and CK20 expression is uncertain in the differential diagnosis between BE and proximal gastric mucosa. On the other hand, positivity for CK7/CK4/CK8 may support the diagnosis of BE and play a role in distinguishing between the two groups. S-100 protein expression and detection of hTERC gene amplification also contribute to the diagnosis of BE.</p>

Clinicopathologic features of papillary tumors of the pineal region / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinicopathologic features of papillary tumor of the pineal region (PTPR).</p><p><b>METHOD</b>Three hundred and eighty six cases of pineal region and posterior third ventricle tumors, two newborn and two adult pineal glands were analyzed by HE, PAS and immunohistochemistry of 16 antibodies (EnVision method).</p><p><b>RESULTS</b>Five cases of PTPR were diagnosed with mixed papillary features and densely cellular areas, and included one recurrent case. In the papillary areas, the vessels were lined by one or several layers of cuboidal/columnar cells; the vessel wall was hyalinized. In the densely cellular areas, sheets or nests of tumor cells were seen. The tumor cells of these five cases were immunoreactive to CK, CK8/18, synaptophysin, MAP2, nestin, S-100, and vimentin. Four cases were immunoreactive to NSE and CgA; and 2 cases were immunoreactive to NF. All five cases were negative for EMA, CK5/6, CEA, and NeuN. Ki-67 labeling index ranged from 1% to 6%.Three patients were alive, and the recurrent one died.</p><p><b>CONCLUSIONS</b>PTPR occurs in patients with over a wide age range, from children to adults, and is more commonly found in male than female. PTPR is composed of both papillary and solid areas, characterized by epithelial cytology, and needs to be differentiated from ependymoma. PTPR may originate from the specialized ependymocytes of the subcommissural organ. The prognostic factors are early diagnosis, complete surgical resection and radiotherapy.</p>

Ethacrynic Acid Inhibits Sphingosylphosphorylcholine-Induced Keratin 8 Phosphorylation and Reorganization via Transglutaminase-2 Inhibition

Sphingosylphosphorylcholine (SPC) is significantly increased in the malicious ascites of tumor patients and induces perinuclear reorganization of keratin 8 (K8) filaments in PANC-1 cells. The reorganization contributes to the viscoelasticity of metastatic cancer cells resulting in increased migration. Recently, we reported that transglutaminase-2 (Tgase-2) is involved in SPC-induced K8 phosphorylation and reorganization. However, effects of Tgase-2 inhibitors on SPC-induced K8 phosphorylation and reorganization were not clearly studied. We found that ethacrynic acid (ECA) concentration-dependently inhibited Tgase-2. Therefore, we examined the effects of ECA on SPC-induced K8 phosphorylation and reorganization. ECA concentration-dependently suppressed the SPC-induced phosphorylation and perinuclear reorganization of K8. ECA also suppressed the SPC-induced migration and invasion. SPC induced JNK activation through Tgase-2 expression and ECA suppressed the activation and expression of JNK in PANC-1 cells. These results suggested that ECA might be useful to control Tgase-2 dependent metastasis of cancer cells such as pancreatic cancer and lung cancers.

Tubulolobular carcinoma of breast: a clinicopathologic study of 8 cases / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinical and morphological features as well as immunophenotype of tubulolobular carcinoma of the breast (TLC).</p><p><b>METHODS</b>Eight cases of TLC were retrieved from 97 cases of invasive lobular carcinoma between January 2005 and March 2010 in the Peking Union Medical College Hospital. The clinical features and pathologic findings were studied and immunohistochemistry was performed for the expression of ER, PR, HER2, p53, E-cadherin, CK34βE12 and CK8.</p><p><b>RESULTS</b>Among the breast cancer patients, the incidence of TLC was about 1.0% (8/880). The mean age of the patients was 59 years, with a range of 45 to 79 years. All patients were asymptomatic, with incidental finding of a mass in the breast on health examination. Common findings on sonography included a hypoechoic nodule with irregular shape and spiculated margin. Histologically, the small uniform tumor cells were arranged in a mixed pattern showing single cells, single-cell files or cords, small round to angulated tubules, and infiltrating lobular or targetoid patterns around ducts that were specific for classical invasive lobular carcinoma. Low or intermediate grade intraepithelial neoplasms which had similar cellular morphology with the invasive tumor often appeared in the periphery, including ductal carcinoma in situ, lobular carcinoma in situ and intraductal papillary carcinoma. Immunohistochemistry of the tumor cells showed intense reactivity to ER (7/8) and PR (8/8), but no reactivity to HER2 or p53. Both the tubules and single-cell file or cords expressed E-cadherin (7/8), CK34βE12 (5/8), and CK8 (8/8) with a uniform staining pattern. All patients underwent modified radical mastectomy and 2/8 patients had metastatic carcinoma in the axillary lymph nodes. Seven patients were followed up for 28 to 75 months and remained well, including one patient that had a new breast mass 60 months after surgery, but had no treatment up to now.</p><p><b>CONCLUSIONS</b>TLC is a rare variant of invasive breast cancer and reveals mixed histologic features of both tubular and lobular carcinoma with common expression of E-cadherin, CK8 and CK34βE12. A better understanding of TLC would enable pathological diagnosis to be made reasonably and accurately.</p>

Experimental study on rat submandibular gland cell and silk fibroin-chitosan in vivo / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the feasibility of the tissue engineered submandibular gland constructed in vivo based on submandibular gland cells and silk fibroin-chitosan (SFCS).</p><p><b>METHODS</b>Submandibular gland cells were obtained and purified. The second generation cells labeled by 5'-BrdU were seeded on SFCS(5 mm × 5 mm × 5 mm). Submandibular gland cells seeded on SFCS was implanted beneath the skin on the back. At 3, 7, 14 d post-implantation, implant sites were examined local and systemic responses. After paraffin embedding, serial sections 6 mm thick were cut and stained with either hematoxylin and eosin or brdu tissue stain for immunohistochemical studies and examined the responses of tissue. Scanning electron microscope was used to observe the growth behavior submandibular gland cells on SFCS scaffolds.</p><p><b>RESULTS</b>General observation: at the 3, 7, 14 d after in vivo implantation, capsule formed in the surface of insert. Histological observation: in experimental group, submandibular gland cells proliferate on the SFCS scaffold fused to form unit 14 d after implantation. Brdu immunohistochemical observation: the results of labelled cells were positive by immunohistochemical method at each time point. Cytokeratin-8 (CK-8) immunohistochemical observation: the results of labelled cells were positive by immunohistochemical method at each time point. With time, the positive cells gradually increased. Scanning electron microscope: the shape of the SFCS scaffold was mesh. At earlier, submandibular gland cells presence in disorder at attach to the SFCS. At the 14 d submandibular gland cells proliferate on the SFCS scaffold and form functional unit.</p><p><b>CONCLUSIONS</b>Such constructed tissue engineered submandibular gland based on submandibular gland cells and SFCS is promising.</p>

Subcellular proteome analysis of immune or alcohol induced rat liver fibrosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the mechanism of liver fibrogenesis and to find new non-invasive biomarkers.</p><p><b>METHOD</b>In this study, we used subcellular proteomic technology to study the plasma membrane proteins related to immune or alcohol induced liver fibrosis. Rat liver fibrosis models were induced by pig serum or alcohol injection. The liver fibrogenesis were detected by James's staining in the rat models after 2, 4, 6 and 8 weeks of treatment. The liver plasma membrane (PM) of the 2- and 8-week treatment model rats were enriched by two-step sucrose density gradient centrifugation. The purity of PM was verified by western blotting, and the plasma membrane proteins were extracted and analyzed by 2 DE. The differentially expressed proteins were identified by LC-MS/MS. Cellular location and function of these identified differential protein were classified.</p><p><b>RESULTS</b>Immune or alcohol induced liver fibrosis rat models were successfully established. Liver plasma membrane was significantly enriched after sucrose density ultracentrifugation treatment. 87 differential protein spots were find out by 2DE combined with LC-MS/MS from the liver plasma membrane proteins of the 2- and 8-week treatment rat models, which corresponded to 30 non-redundant proteins including annexin A2, keratin 8 and keratin 18.</p><p><b>CONCLUSIONS</b>A list of differentially expressed proteins relate to liver fibrosis were successfully identified. Differential proteins such as annexin A2, keratin 8 and keratin 18 could be new biomarkers for liver fibrosis diagnosis.</p>

Expression of keratin 8 in carbon tetrachloride-induced liver injury of mice / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the expression of keratin 8 (K8) in carbon tetrachloride (CCl(4))-induced liver injury of mice.</p><p><b>METHODS</b>Forty ICR mice were divided into four groups. CCl(4) 300 microl/kg body weight in olive oil was injected intraperitoneally for 0, 2, 4, 6 weeks in group A, B, C and D, respectively. Mice were sacrificed 3 d after the last injection and then the vital organs were collected and weighed. RT-PCR and immunohistochemistry methods were used to analyze the expression of keratin 8 in the liver.</p><p><b>RESULTS</b>The ratios of liver and body weight were increased significantly after administration of CCl(4), which were 5.60 %, 6.87%, 7.83 % and 7.76% at 0, 2, 4 and 6 weeks after injection, respectively. The expression of K8 was increased at the 2 w, 4 w and 6 w after CL(4) administration.</p><p><b>CONCLUSION</b>The expression of K8 is positively correlated with the liver injury induced by CCl(4). The accumulation of K8 may be involved in the mechanism of liver injury.</p>

Time-course Transcriptional Profiling of Human Amniotic Fluid-derived Stem Cells Using Microarray / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: To maintain the homeostasis of stem cells and prevent their ability to initiate tumorigenesis, it is important to identify and modify factors that prevent or accelerate stem cell senescence. We used microarrays to attempt to identify such factors in human amniotic fluid (HAF)-derived stem cells. MATERIALS AND METHODS: To identify gene expression changes over a time course, we compared gene expression profiles of HAF-derived stem cells in different passages (1st, 2nd, 4th, 6th, 8th, and 10th) using a Sentrix Human illumina microarray. RESULTS: Of the 25,804 genes in the microarray chip, 1,970 showed an over 2-fold change relative to the control (the 1st passage)-either upregulated or downregulated. Quantitative real-time PCR validated the microarray data for selected genes: markedly increased genes were CXCL12, cadherin 6 (CDH6), and folate receptor 3 (FOLR3). Downregulated genes included cyclin D2, keratin 8, insulin-like growth factor 2 (IGF2), natriuretic peptide precursor B (NPPB) and cellular retinoic acid binding protein 2 (CRABP2). The expression pattern of the selected genes was consistent with the microarray data except for CXCL12 and IGF2. Interestingly, the expression of NPPB was dramatically downregulated along the time course; it was almost completely shut-down by the 10th passage. In contrast, FOLR3 mRNA expression was dramatically increased. CONCLUSION: Taken together, although a function for NPPB and FOLR3 in stem cell senescence has not been reported, our results strongly suggest that NPPB and/or FOLR3 play a significant role in the regulation of stem cell senescence.

Proteomic study of paclitaxel on human cervical carcinoma HCE1 / 中南大学学报(医学版)

OBJECTIVE@#To explore the mechanism of paclitaxel on the protein expression of human cervical carcinoma cell line HCE1.@*METHODS@#The total proteins extracted from paclitaxel-treated HCE1 cells were analyzed by 2-dimensional gel electrophoresis (2-DE), and compared with those from untreated HCE1 cells. The differential proteins were identified by mass spectrometry. Western blot was used to determine the differential expression levels of the 2 proteins.@*RESULTS@#At 24 hour after paclitaxel (0.05 mumol/L) treatment, 2-DE images of paclitaxel-treated and paclitaxel-untreated cells were analyzed. Forty-two differential proteins were found. Twenty-one differential proteins among 42 proteins were analyzed by mass spectrometry, among which 15 proteins were identified, including peptidyl-prolylisomerases A (PPIase A),alpha-enolase,keratin 8,heat shock protein 90, eukaryotic translation initiation factor 1A, and so on.@*CONCLUSION@#Fifteen proteins in human cervical carcinoma cells paclitaxel-treated and paclitaxel-untreated are found by proteomic techniques. These proteins may be involved in the proliferation inhibition of human cervical carcinoma cells by paclitaxel.

Determination of differentially expressed proteins and it's significance among chronic sinusitis, nasal polyps and normal nasal mucosa / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the differentially expressed proteins among chronic sinusitis, nasal polyps and normal nasal mucosa by means of proteomic technology, and select the candidate biomarkers of chronic sinusitis and nasal polyps.</p><p><b>METHODS</b>Proteins extracted from chronic sinusitis, nasal polyps and normal nasal mucosa were separated and the differentially expressed proteins were identified by series of proteomic tools, including immobilized pH4-7 gradient two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis, modified coomassie brilliant blue staining, images scanning by the Image Scanner apparatus, PDQuest analysis software, peptide mass fingerprinting based on matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) by in-gel digestion extract, and Mascot searching in NCBInr and SWISS-PROT databases.</p><p><b>RESULTS</b>The 2-DE patterns with high resolution and reproducibility were obtained. The protein spots separated and visualized in chronic sinusitis, nasal polyps and normal nasal mucosa gel were 1020 +/- 40, 1112 +/- 10 and 1008 +/- 25, respectively. And the match rates were (93 +/- 2)%, (95 +/- 1)% [see text] (90 +/- 3)% respectively. Thirteen differentially expressed spots were found from chronic sinusitis, nasal polyps and normal nasal mucosa gel. We selected and recommend Keratin 8 and APOA1 proteins as candidate biomarkers of nasal polyps, and PLUNC protein, PACAP protein, NKEF-B and SOD as candidate biomarkers of chronic sinusitis.</p><p><b>CONCLUSIONS</b>The differentially expressed proteins among chronic sinusitis, nasal polyps and normal nasal mucosa can be efficiently and relatively reliably identified via the techniques of proteomics. These techniques will play a very important role in the researches for new objective indicators possibly employed in the future classifying, staging and prognosis.</p>

Cytokeratin Autoantibodies: Useful Serologic Markers for Toluene Diisocyanate-Induced Asthma

To evaluate the clinical significance of autoantibodies to three major epithelial cytokeratins (CK) -- CK8, CK18, and CK19 -- we compared 66 patients with toluene diisocyanate (TDI)-induced asthma (group I) with three control groups: 169 asymptomatic exposed subjects (group II), 64 patients with allergic asthma (group III), and 123 unexposed healthy subjects (group IV). Serum IgG, specific for human recombinant CKs, were measured by ELISA (enzyme linked immunosorbent assay), and ELISA inhibition tests were performed. The existence of these antibodies was confirmed by IgG immunoblot analysis. Anti-TDI-HSA (human serum albumin) IgE and IgG antibodies were measured by ELISA in the same set of the patients. The prevalence of CK8, CK18, and CK19 auotantibodies in group I was significantly higher than in the other three groups. Results of the ELISA inhibition test showed significant inhibition with the addition of three CKs in a dose-dependent manner. No significant association was found between CK autoantibodies and the prevalence of anti- TDI-HSA IgG and IgE antibodies. These results suggest that autoantibodies to CK18 and CK19 can be used as serologic markers for identifying patients with TDI-induced asthma among exposed workers.

Proteomic analysis of paclitaxel-induced apoptosis in MCF-7 human breast carcinoma cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the mechanism of paclitaxel-induced apoptosis in MCF-7 human breast carcinoma cells.</p><p><b>METHODS</b>In this study, the proteins extracted from paclitaxel-induced apoptotic MCF-7 cells were analyzed by 2-dimentional gel electrophoresis (2-DE), and compared with those from untreated MCF-7 cells. The differential proteins were identified by mass spectrometry.</p><p><b>RESULTS</b>At 24 hour after paclitaxel (100 nmol/L) treatment, MCF-7 cells were collected and extracted the whole proteins. Seventeen up-regulated or down-regulated proteins were found by analysis of the differential proteomic 2-DE map. Six of them were identified by mass spectrometry. They were enolase 1, chloride intracellular channel 1, keratin 8, ribosomal protein S12, galectin-1 and histidine triad nucleotide binding protein, respectively.</p><p><b>CONCLUSION</b>We effectively found the changed proteins in the process of paclitaxel-induced apoptosis in MCF-7 human breast carcinoma cells by proteomic techniques. These up-regulated or down-regulated proteins are important molecules for our further research about the mechanism of paclitaxel-induced apoptosis.</p>

Clinicopathologic study of solid papillary carcinoma of breast / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinicopathologic features and immunophenotype of solid papillary carcinoma (SPC) of breast.</p><p><b>METHODS</b>Clinical and pathologic features of 21 cases of SPC, with or without stromal invasion, were analyzed. Immunohistochemical study (LSAB method, for cytokeratins, myoepithelial markers, chromogranin A, synaptophysin, Ki-67, estrogen receptor, progesterone receptor, c-erbB-2 and pS2) and alcian blue staining were performed.</p><p><b>RESULTS</b>All the patients were females with a mean age of 66.1 years. The clinical features were similar to those of classic papillary tumor. Metastasis was not observed in patients who had undergone axillary lymph node dissection. Histologically, the tumor displayed solid papillary growth pattern, with mucin production demonstrated in 19 cases. The tumor cells were oval, polygonal, spindled or signet ring-like and contained mildly to moderately pleomorphic nuclei. The mitotic count measured less than 5 per 10 high-power fields in 15 cases. Seven cases contained foci of invasive carcinoma which showed similar cytologic features as those of the in-situ component. Immunohistochemical study showed that the tumor cells expressed CK8 but not basal cell cytokeratin. Positivity for smooth muscle actin-alpha, calponin and p63 was demonstrated in the myoepithelial layers of fibrovascular cores, as well as around the expanded ductolobular units. Most cases also showed cytoplasmic positivity for chromogranin A (88.2%) and synaptophysin (82.4%). The proliferation index, as highlighted by Ki-67 immunostain, was 8.1%. The tumor expressed estrogen receptor, progesterone receptor and pS2. The staining for c-erbB-2 oncoprotein was negative. Follow up of 16 patients showed no evidence of recurrence or metastasis.</p><p><b>CONCLUSIONS</b>SPC predominantly affects elderly females and has distinctive pathologic features and immunophenotype. Some cases of SPC are associated with mucinous and neuroendocrine components. Follow-up data suggest that SPC often carries an indolent clinical behavior and favorable prognosis.</p>

Effect of 12-O-tetradecanoylphorbol 13-acetate (TPA) on the Expression of Keratin in HaCaT Cells / 대한해부학회지

In human skin, specific keratin markers reflect on normal differentiation and pathologic conditions. This experiment focused on the expressional pattern of keratin 10 (K10: normal differentiation marker), and keratin 8 & 13 (K8 & K13: pathologic differentiation marker) together with their cellular localization after treating HaCaT cells with 12-Otetradecanoylphorbol 13-acetate (TPA). The cells were treated with TPA at 0, 0.1, 1 microgram/ml for 2 hours or 6 hours. Morphologic studies revealed that TPA treatment changed the shape of cells into the fibroblast-like cells with highly folded nuclear membrane and reduced number of the desmosome. The results of indirect immunofluorescent staining and Northern blotting analysis showed that TPA considerably down-regulated the expression of K10, while markedly up-regulating the expression of K8 and K13 both at protein and mRNA levels. Furthermore, by simultaneous staining for keratins and DNA content in flow cytometry, it was found that TPA increased the expression of K8 and K13 dramatically at the S-G2-M phase of the cells. In conclusion, these changes induced by TPA in HaCaT cells may indicate a close relationship between the morphologic change and the altered expression of keratin subfamilies. It also suggests that TPA known as a tumor promotor may directly induce the potentially malignant cells even without the support of tumor initiator.

Co-relationship between Expression of Keratins and Vimentin in Breast Cancer Tissures and Metastases of Breast Cancer / 한국유방암학회지

PURPOSE: the most important biological behavior of breast cancer is its invasive potential and many efforts was made to reveal the factors related with the invasiveness of breast cancer cells. Some researchers reported that intermediate filament biology could represent an emerging and exciting field in tumor biology with respect to tumor aggressiveness and invasiveness. There are some experimental evidences that co-expression of vimentin, a interfilament marker indicative of mesenchymal lineage, and cytokeratin interfilaments can be correlated with invasiveness and metastatic deposits. So, we tried to determine the role of intermediate filaments such as cytokeratins and vimentin with respect of bone marrow micrometastases. METHODS: Expression of cytokeratins 8, 18, 19 and viementin were immunohistochemically evaluated. Detection of bone marrow micrometastases was preformed through RT-PCR targeting mRNA of cytokeratin 19. In order to compare the study group by the expression extent of cytokeratins, the case expressing 50% or more of observed cells was classified into the case with high expression and the case expressing 49% or less was classified into the case with low expression. RESULTS: The only cytokeratin of high expression representing the risk of bone marrow micrometastases was cytokeratin 8. Vimentin expression by itself did not show any significance indicating bone marrow micrometastases. However, The cases possesing cytokeratin 8, 18, and 19 expression, altogether 75% or more showed a significantly high risk to bone marrow micrometastases. In that cases, addition of vimentin expression allowed a more higher possibility of bone marrow micrometastases. CONCLUSION: A high expression of cytokeratin 8 among cytokeratins was related with bone marrow metastases. However, vimentin expression by itself did not show any realtionship with bone marrow metastases. So, a further study is needed in order to reveal the role of vimentin expression in progression and metastases of breast cancer.

Diagnostic Value of Serum Cytokeratin 8, 18 and 19 in Lung Cancer / 결핵

BACKGROUND: Monoclonal antibodies directed against well-known epitopes on cytokeratin (CK) 8, 18 and 19 (Monototal) have been used in the development of a new diagnostic tool for lung cancer. In the mid-1990s, CK 19 fragments (Cyfra 21-1) became popular and widely used for such diagnosis. This is the first study specifically designed to compare these two markers. METHOD: The serum levels of CK 8, 18 and 19 were measured using two-site monoclonal/polyclonal immunoradiometric assay kit in 57 healthy adults and 289 patients who were admitted to Seoul National University Hospital from May to September, 2002. The lung cancer group comprised 129 primary lung cancer patients; 116 with non-small cell lung cancer(NSCLC) and 13 with small cell lung cancer (SCLC). The control group comprised 160 non-malignant pulmonary lung disease patients and 57 healthy adults. A total of 166 twin Monototal and Cyfra 21-1 serum assays were obtained; 76 with lung cancer, 70 with non-malignant pulmonary lung disease and 20 healthy adults. RESULTS: The mean serum value of Monototal was 412.47+/-455.45U/L in NSCLC, 237.08+/-145.15U/L in SCLC, 126.54+/-95.72U/L in non-malignant pulmonary lung disease, and 63.68+/-31.66U/L in healthy adults. The serum values of the lung cancer groups were significantly higher than those of the control group (p<0.01). Using a cut off value of 188U/L, sensitivity and specificity was 66.4% and 81.9% in NSCLC, and 43.8% and 81.9% in SCLC, respectively. The serum levels of CK 8, 18 and 19 were higher in advanced NSCLC than in early stage disease. CONCLUSION: The serum levels of CK 8, 18 and 19 may be useful in the diagnosis of NSCLC.

Study on the relationship between epidermal stem cells and the developing process of sweat gland in human fetal skin / 中华烧伤杂志

<p><b>OBJECTIVE</b>To explore the relationship between epidermal stem cells and the developing process of sweat gland in human fetal skin, so as to obtain a hint for future induction of epidermal stem cells to differentiate into sweat gland cells.</p><p><b>METHODS</b>Total layer of human skin from the back of fetus at gestational ages from 11 to 31 weeks, obtained from spontaneous abortion was routinely examined. The expressions of beta1 integrin and keratin 19 in sweat gland cords or buds and mature sweat gland cells were dynamically observed with SP immunohistochemical technique. The development and maturation of sweat gland were identified by the positive staining of keratin 8 with immunohistochemistry.</p><p><b>RESULTS</b>It was revealed by histologic observation that basal layer cells of the primary epidermal ridge exhibited focal aggregation and formed hillocks at 16 gestational weeks. The hillocks of cells then migrated downward as cords into the dermis during 18 - 20 gestational weeks. Then, the end part of the cell cord developed into a round lump of twining cords assuming the mature sweat gland. The expressions of beta1 integrin and keratin 19 were found not only in sweat gland cords and buds but also in the mature cells and lasted throughout the total period of sweat gland development. The expression of keratin 8 in sweat gland buds started since 14 - 16 gestational weeks and maintained thereafter.</p><p><b>CONCLUSION</b>The sweat gland started to develop during 14 - 16 gestational weeks and matured at 24 weeks. During the development process of sweat gland, epidermal stem cells were considered to be the key source.</p>

Clinical Significance of the Urinary Bladder Cancer Antigen (UBCTM) Test in the Diagnosis of Bladder Cancer / 대한비뇨기과학회지

PURPOSE: A new quantitative tumor marker, based on the combined measurement of urinary fragments of cytokeratin 8 and 18, namely the urinary bladder cancer antigen (UBCTM) test, has been proposed for the detection of bladder cancer. We compared the results of the UBC test, with voided urine cytology, for the diagnosis of bladder cancer to evaluate its diagnostic performance. MATERIALS AND METHODS: The UBC concentrations were measured, using an immunoradiometric assay, in the urine of 15 healthy subjects (group I), 26 patients with other urological disease except bladder cancer (group II), 40 patients with active bladder cancer (group III) and 17 patients free of bladder cancer, as confirmed by cystoscopy at follow-up (group IV). The differences in the UBC test, with regard to stage, grade, tumor size, focality and history of recurrence, were also evaluated. RESULTS: The mean UBC concentrations were 3.52micro gram/l, 45.76micro gram/l, 92.80micro gram/l and 20.51micro gram/l, for group I to IV, respectively, which were statistically different (p<0.05). There were significant differences regarding stage (p=0.044) and tumor size (p=0.036). However, no differences were founded in relation to the grade, shape, focality or history of recurrence. The optimal threshold for the UBC test, and the area under the ROC curve, were 12.8micro gram//l and 0.684, respectively. The sensitivity, specificity, positive and negative predictive values for the UBC test and urine cytology in groups III and IV were 50.0 and 30.0%, 88.2 and 100%, 90.9 and 100%, and 42.9 and 37.0%, respectively. CONCLUSIONS: The UBC test appears to be useful for the detection of bladder cancer in terms of its superior sensitivity and negative predictive value over those of urine cytology. Further studies will be required for the clinical utility of the UBC test in the diagnosis and follow-up of bladder cancer.