Objective To screen antigen targets for immunotherapy by analyzing over-expressed genes, and to identify significant pathways and molecular mechanisms in esophageal cancer by using bioinformatic methods such as enrichment analysis, protein-protein interaction (PPI) network, and survival analysis based on the Gene Expression Omnibus (GEO) database.Methods By screening with highly expressed genes, we mainly analyzed proteins MUC13 and EPCAM with transmembrane domain and antigen epitope from TMHMM and IEDB websites. Significant genes and pathways associated with the pathogenesis of esophageal cancer were identified using enrichment analysis, PPI network, and survival analysis. Several software and platforms including Prism 8, R language, Cytoscape, DAVID, STRING, and GEPIA platform were used in the search and/or figure creation.Results Genes MUC13 and EPCAM were over-expressed with several antigen epitopes in esophageal squamous cell carcinoma (ESCC) tissue. Enrichment analysis revealed that the process of keratinization was focused and a series of genes were related with the development of esophageal cancer. Four genes including ALDH3A1, C2, SLC6A1,and ZBTB7C were screened with significant P value of survival curve.Conclusions Genes MUC13 and EPCAM may be promising antigen targets or biomarkers for esophageal cancer. Keratinization may greatly impact the pathogenesis of esophageal cancer. Genes ALDH3A1, C2, SLC6A1,and ZBTB7C may play important roles in the development of esophageal cancer.
Humans ; Esophageal Neoplasms/metabolism* ; Esophageal Squamous Cell Carcinoma/metabolism* ; Epithelial Cell Adhesion Molecule/metabolism* ; Gene Expression Profiling/methods* ; Gene Regulatory Networks ; Gene Expression ; Gene Expression Regulation, Neoplastic ; Intracellular Signaling Peptides and Proteins

PURPOSE: Epithelial barrier dysfunction is involved in the pathophysiology of periodontitis and oral lichen planus. Estrogens have been shown to enhance the physical barrier function of intestinal and esophageal epithelia, and we aimed to investigate the effect of estradiol (E2) on the regulation of physical barrier and tight junction (TJ) proteins in human oral epithelial cell monolayers. METHODS: HOK-16B cell monolayers cultured on transwells were treated with E2, an estrogen receptor (ER) antagonist (ICI 182,780), tumor necrosis factor alpha (TNFα), or dexamethasone (Dexa), and the transepithelial electrical resistance (TER) was then measured. Cell proliferation was measured by the cell counting kit (CCK)-8 assay. The levels of TJ proteins and nuclear translocation of nuclear factor (NF)-κB were examined by confocal microscopy. RESULTS: E2 treatment increased the TER and the levels of junctional adhesion molecule (JAM)-A and zonula occludens (ZO)-1 in a dose-dependent manner, without affecting cell proliferation during barrier formation. Treatment of the tight-junctioned cell monolayers with TNFα induced decreases in the TER and the levels of ZO-1 and nuclear translocation of NF-κB. These TNFα-induced changes were inhibited by E2, and this effect was completely reversed by co-treatment with ICI 182,780. Furthermore, E2 and Dexa presented an additive effect on the epithelial barrier function. CONCLUSIONS: E2 reinforces the physical barrier of oral epithelial cells through the nuclear ER-dependent upregulation of TJ proteins. The protective effect of E2 on the TNFα-induced impairment of the epithelial barrier and its additive effect with Dexa suggest its potential use to treat oral inflammatory diseases involving epithelial barrier dysfunction.
Architectural Accessibility ; Cell Count ; Cell Proliferation ; Dexamethasone ; Electric Impedance ; Epithelial Cells ; Estradiol ; Estrogens ; Humans ; Junctional Adhesion Molecule A ; Junctional Adhesion Molecules ; Lichen Planus, Oral ; Microscopy, Confocal ; NF-kappa B ; Periodontitis ; Tight Junctions ; Tumor Necrosis Factor-alpha ; Up-Regulation

OBJECTIVE: To investigate clinicopathological and prognostic significance of epithelial cell adhesion molecule (EpCAM) expression in breast cancer and its molecular subtypes.
 METHODS: The expression of EpCAM in 835 patients with breast invasive ductal carcinoma was detected by immunohistochemical Max VisionTM method, and its correlation with clinical pathological features and prognosis was analyzed.
 RESULTS: The positive expression of EpCAM was related to histological grade, lymph node metastasis, tumor size, clinical stage, the expression of estrogen receptor (ER), progesterone receptor (PR) and HER2 (P<0.05). The positive expression rates of EpCAM were 19.2%, 73%, 48.9%, 72.2%, and 62.1%, in Luminal A, Luminal B (HER2-), Luminal B(HER2+), HER2+, and triple-negative subtype, respectively. Log-rank test and univariate COX analysis showed that the EpCAM expression was associated with a poor prognosis in all patients (P<0.001), as well as the triple-negative subtype, luminal B subtype (HER2-), and HER2+ subtype (P<0.05). Multivariate COX analysis showed that EpCAM expression was associated with the survival in patients with the triple-negative or HER2+ subtype (P<0.05).
 CONCLUSION EpCAM may be associated with progress of breast cancer. It is an independent prognostic factor in breast cancer, especially in the triple-negative and HER2+ subtypes.
Breast Neoplasms ; Epithelial Cell Adhesion Molecule ; Humans ; Lymphatic Metastasis ; Prognosis ; Receptor, ErbB-2 ; Receptors, Progesterone

To examine the expressions of epithelial cell adhesion molecule (EpCAM), vimentin and N-cadherin in breast cancer and its molecular subtypes, and to explore the correlation among them.
 Methods: The expressions of EpCAM, vimentin and N-cadherin were detected by immunohistochemistry in 835 patients with breast cancer, and their correlations with clinical pathological features and prognosis were analyzed.
 Results: The expression rates of EpCAM, vimentin and N-cadherin in the patients were 53.4%, 11.4% and 9.7% respectively, which were increased with the increase in tumor size, histological grade, lymph node size, tumor node stage of metastases classification, estrogen receptor and progesterone receptor levels (all P<0.05). The positive expression rates for EpCAM protein in luminal A, luminal B (HER2-), luminal B (HER2+), HER2 overexpression and triple-negative subtypes were 19.2%, 73.0%, 48.9%, 72.2%, and 62.1% respectively; for vimentin were 3.9%, 11.4%, 14.1%, 11.1%, and 20.5% respectively; for N-cadherin were 7.0%, 5.7%, 12.0%, 12.2% and 17.4% respectively, with statistical difference (all P<0.05). EpCAM expression was positively correlated with vimentin and N-cadherin in patients with breast cancer and triple-negative breast cancer.
 Conclusion: EpCAM is overexpressed in triple-negative subtype of breast cancer and it is associated with epithelial-mesenchymal transition markers, which might be related to breast cancer progression and metastasis.
Antigens, CD ; Biomarkers, Tumor ; Breast Neoplasms ; chemistry ; classification ; genetics ; Cadherins ; Epithelial Cell Adhesion Molecule ; Epithelial-Mesenchymal Transition ; Female ; Humans ; Immunohistochemistry ; Neoplasm Grading ; Neoplasm Staging ; Prognosis ; Receptor, ErbB-2 ; Receptors, Estrogen ; Receptors, Progesterone ; Triple Negative Breast Neoplasms ; Vimentin

OBJECTIVETo compare the results of pathological diagnosis of 41 patients with malignant mesothelioma between Chinese and Japanese experts, and to provide a basis for the standard for diagnosis of mesothelioma.

METHODSThe medical information and tissue samples of 41 patients with malignant mesothelioma were collected in a hospital in Zhejiang Province from 2003 to 2010. The expression levels of calretinin, Wilms' tumor suppressor gene (WT1), podoplanin (D2-40), cytokeratins (CK5/6, AE1/AE3, and CAM5.2), epithelial membrane antigen, carcinoembryonic antigen, BerEP4, MOC31, thyroid transcription factor-1, estrogen receptor, and progesterone receptor in tumor tissues were measured using immunohistochemical staining by Japanese experts, and the pathological classification and diagnosis were made. The results of diagnosis, pathological classification, immunohistochemical marker selection, and slide review were compared between Chinese and Japanese experts.

RESULTSTwenty-nine (70.7%) cases were diagnosed as mesothelioma by Japanese experts, among whom 12 (41.4%) cases were pleura mesothelioma, and 17 (58.6%) cases were peritoneal mesothelioma. Ten (24.4%) cases were confirmed without mesothelioma, and 2 (4.9%) cases were not confirmed due to insufficient information. Thirty-two (78.0%) cases were diagnosed as mesothelioma by Chinese experts, among whom 8 (25.0%) cases were pleura mesothelioma, and 24 (75.0%) cases were peritoneal mesothelioma. One (2.4%) case was confirmed without mesothelioma, and 8 (19.5%) cases were not confirmed. There were significant differences in the results of diagnosis between Chinese and Japanese experts. However, their pathological classifications of mesothelioma were similar. Significant differences in immunohistochemical marker selection and slide review were also found between Chinese and Japanese experts.

CONCLUSIONThe diagnostic skills of those pathological experts in this hospital remain to be further improved for mesothelioma diagnosis. A panel of immunohistochemical markers including at least 2 mesothelioma-positive and 2 mesothelioma-negative markers are recommended for the diagnosis of malignant mesothelioma.

Antigens, Neoplasm ; Biomarkers ; Biomarkers, Tumor ; Cell Adhesion Molecules ; China ; Diagnostic Techniques and Procedures ; standards ; Epithelial Cell Adhesion Molecule ; Humans ; Immunohistochemistry ; Japan ; Lung Neoplasms ; classification ; diagnosis ; Mesothelioma ; classification ; diagnosis ; Observer Variation

OBJECTIVETo investigate the transformative potential of hepatic progenitor cells to differentiate into liver stem cells using a normal adult mouse system and to determine the effects of HBx protein in these liver stem cells' differentiation into hepatic cells.

METHODSHepatic progenitor cells were obtained from mice by means of an optimized two-step digestion and perfusion method followed by joint differential centrifugation and density gradient centrifugation. Transformation of the hepatic progenitor cells into liver stem cells was observed by immunofluorescent detection of CD 133, EPCAM, CD49f and CK19. Differentiation of the resultant liver stem cells into hepatic cells and bile duct epithelial cells was observed after DMSO addition by Periodic Acid-Schiff (PAS) staining followed by cell immunofluorescence and flow cytometry. To determine the effects of HBx on these liver stem cells' ability to differentiate into hepatic cells, cell transfection was used followed by observation of morphology and proliferation capacity.

RESULTSCell viability of the isolated hepatic progenitor cells was 78.67+/-4.04%. Stimulation with EGF and collagen led to growth of some of the paving-stone shaped cells attached to the hepatic progenitor cells which had gathered into spherical clumps, as is the nature of stem cells. The liver stem cells showed high expression of CD133, CD49f and CK19, and low expression of EPCAM. Under the effect of DMSO, the liver stem cells differentiated into hepatocytes and bile duct epithelial cells. After HBx transfecfion, the liver stem cells maintained the characteristic shape of stem cells and showed enhanced proliferation.

CONCLUSIONEPCAM-positive adult hepatic progenitor cells can transform into liver stem cells.The HBx protein may play an important role in maintaining the stability of liver stem cells in the adult mouse.

Animals ; Antigens, Neoplasm ; metabolism ; Bile Ducts ; cytology ; Cell Adhesion Molecules ; metabolism ; Cell Differentiation ; Epithelial Cell Adhesion Molecule ; Epithelial Cells ; cytology ; Flow Cytometry ; Hepatocytes ; cytology ; Liver ; cytology ; Mice ; Stem Cells ; cytology

OBJECTIVETo explore the expression of the epithelial cell adhesion molecule (EpCAM) in prostate cancer (PCa) and its clinical significance.

METHODSWe collected tissue samples from 63 cases of PCa, 46 cases of prostatic intraepithelial neoplasia (PIN), and 58 cases of benign prostatic hyperplasia (BPH) adjacent to PCa and determined the expression of EpCAM in the epithelial and stromal cells by immunohistochemistry.

RESULTSThe positive expression rates of EpCAM in the epithelial cells were significantly higher in PCa and PIN than in PCa-adjacent BPH (98. 4 and 97. 8 vs 51.7%, P <0. 01), and so was that in the stromal cells of PCa than in those of PCa-adjacent PIN (89.5 vs 50.0%, P <0.01). The expression of EpCAM.was remarkably higher in the stromal cells of bone metastasis than in those of non-bone metastasis tissue (100. 0 vs 40. 0%, P <0. 01) but showed no statistically significant differences between the highly and poorly differentiated PCa tissues (88.5 vs 91.9%, P >0.05).

CONCLUSIONThe expression level of EpCAM in the stromal cells of PCa is related to the occurrence, progression, and bone metastasis of the tumor, and therefore may be used as a marker in the early diagnosis of PCa as well as a predictor of bone metastasis of the tumor.

Antigens, Neoplasm ; metabolism ; Biomarkers ; metabolism ; Bone Neoplasms ; metabolism ; secondary ; Cell Adhesion Molecules ; metabolism ; Disease Progression ; Epithelial Cell Adhesion Molecule ; Epithelial Cells ; metabolism ; Humans ; Immunohistochemistry ; Male ; Prostatic Hyperplasia ; metabolism ; Prostatic Intraepithelial Neoplasia ; metabolism ; Prostatic Neoplasms ; metabolism ; Stromal Cells ; metabolism

OBJECTIVETo study the expression of EpCAM and E-cadherin in papillary thyroid carcinoma and to analyze its correlation with various clinicopathologic parameters.

METHODSImmunohistochemical study for EpCAM and E-cadherin was carried out in 91 cases of papillary thyroid carcinoma. Twenty-four cases of papillary hyperplasia of thyroid were used as controls.

RESULTSIn all of the 24 cases of papillary hyperplasia, EpCAM was located on the cell membrane, while in the 91 cases of papillary thyroid carcinoma studied, EpCAM was located within the cytoplasm, with 36.3% (33/91) showing nuclear localization as well. In all the papillary hyperplasia cases studied, E-cadherin showed membranous expression. E-cadherin expression was reduced in 84.6% (77/91) of papillary thyroid carcinoma, as compared with the surrounding native thyroid parenchyma. Amongst the 33 cases of papillary thyroid carcinoma which showed nuclear localization of EpCAM, 30 cases also showed reduced E-cadherin expression. There was a positive correlation between nuclear expression of EpCAM and loss of E-cadherin expression (P = 0.000; Spearman correlation coefficient = 0.857). Nuclear expression of EpCAM correlated with follicular variant of papillary thyroid carcinoma and presence of extrathyroidal extension ( P = 0.037 and 0.033, respectively). Loss of E-cadherin expression correlated with age of patients and presence of lymph node metastasis (P = 0.018 and 0.010, respectively).

CONCLUSIONSE-cadherin expression is reduced in papillary thyroid carcinoma, as compared with native thyroid parenchyma and papillary hyperplasia. Papillary thyroid carcinoma shows loss of EpCAM membranous expression and increased cytoplasmic/nuclear accumulation. Detection of these two markers may provide a valuable reference in defining the biologic behaviors of papillary thyroid carcinoma, including extrathyroidal extension and lymph node metastasis.

Antigens, Neoplasm ; metabolism ; Cadherins ; metabolism ; Carcinoma, Papillary ; metabolism ; secondary ; Cell Adhesion Molecules ; metabolism ; Cell Membrane ; metabolism ; Cytoplasm ; metabolism ; Epithelial Cell Adhesion Molecule ; Humans ; Lymphatic Metastasis ; Neoplasm Proteins ; metabolism ; Thyroid Neoplasms ; metabolism ; pathology

OBJECTIVETo study the cell morphology change in dormancy and proliferation stage of colorectal cancer stem cells in order to provide reference to the treatment of colorectal cancer.

METHODSThe subpopulation of EpCAM(high)/CD44(+)/CD133(+) was isolated from fresh colorectal cancer tissues. These cells were tested by xenograft assay in NOD/SCID nude mice. Colorectal cancer stem cells underwent three-dimensional culture, and the growth curve of stem cells was drawn by WST-1. The expression of P27 and Ki-67 was examined by flow cytometry to understand the phase of dormancy and proliferation of colorectal cancer stem cells. Then the morphological differences of colorectal cancer stem cells between dormant and proliferation stages were recognized by immunofluorescence staining of actin.

RESULTSThe percentage of EpCAM(high)/CD44(+)/CD133(+) was 1.6%, and the subpopulation was confirmed to be colorectal cancer stem cells by means of the experiment of tumorigenicity in vivo. The growth curve of colorectal cancer stem cells was "S" type. Colorectal cancer stem cells grew slowly in the first three days. The expression of P27 was gradually up-regulated, and the level of Ki-67 was very low. These cells remained quiescence, which was the so-called dormancy. The expression of Ki-67 of colorectal cancer stem cells was at high level since the fourth day, and the P27 level was very low. According to the growth curve, this period belonged to the proliferative stage of colorectal cancer stem cells. On immunofluorescence staining, colorectal cancer stem cells with high level of P27 were round, large, and few pseudopodium, but no obvious death was found. These cells showed characteristics of dormancy. In contrast, the stem cells with high level of Ki-67 had much pseudopodium, showing proliferation and invasion.

CONCLUSIONSCancer recurrence and metastasis may be associated with the change of growth state of cancer stem cells. Colorectal cancer stem cells in the proliferation stage show greater proliferative and invasive ability as compared to the dormancy stage, which provides a new perspective for the treatment of colorectal cancer, and recurrence and metastasis of other tumors.

Animals ; Antigens, CD ; Antigens, Neoplasm ; Cell Adhesion Molecules ; Cell Cycle Checkpoints ; Cell Proliferation ; Cell Shape ; Colorectal Neoplasms ; Epithelial Cell Adhesion Molecule ; Glycoproteins ; Mice ; Mice, Inbred NOD ; Mice, Nude ; Mice, SCID ; Neoplasm Recurrence, Local ; Neoplastic Stem Cells ; cytology

PURPOSE: Lovastatin is an effective inhibitor of cholesterol synthesis. A previous study demonstrated that lovastatin can also suppress airway hyperresponsiveness (AHR) in murine model of asthma. We aimed to investigate the effect of lovastatin on mucus secretion and inflammation-associated gene expression in the lungs of murine model of asthma. METHODS: Female BALB/c mice were sensitized and challenged with ovalbumin (OVA) by intraperitoneal injection, and orally administered lovastatin from days 14 to 27 post-injection. Gene expression in lung tissues was analyzed using real-time polymerase chain reaction. AHR and goblet cell hyperplasia were also examined. BEAS-2B human bronchial epithelial cells were used to evaluate the effect of lovastatin on the expression of cell adhesion molecules, chemokines, and proinflammatory cytokines in vitro. RESULTS: We showed that lovastatin inhibits the expression of Th2-associated genes, including eotaxins and adhesion molecules, in the lungs of murine model of asthma. Mucin 5AC expression, eosinophil infiltration and goblet cell hyperplasia were significantly decreased in the lung tissue of murine model of asthma treated with lovastatin. Furthermore, lovastatin inhibited AHR and expression of Th2-associated cytokines in bronchoalveolar lavage fluid. However, a high dose (40 mg/kg) of lovastatin was required to decrease specific IgE to OVA levels in serum, and suppress the expression of Th2-associated cytokines in splenocytes. Activated BEAS-2B cells treated with lovastatin exhibited reduced IL-6, eotaxins (CCL11 and CCL24), and intercellular adhesion molecule-1 protein expression. Consistent with this, lovastatin also suppressed the ability of HL-60 cells to adhere to inflammatory BEAS-2B cells. CONCLUSIONS: These data suggest that lovastatin suppresses mucus secretion and airway inflammation by inhibiting the production of eotaxins and Th2 cytokines in murine model of asthma.
Animals ; Asthma* ; Bronchoalveolar Lavage Fluid ; Cell Adhesion Molecules ; Chemokines ; Cholesterol ; Cytokines ; Eosinophils ; Epithelial Cells ; Female ; Gene Expression ; Goblet Cells ; HL-60 Cells ; Humans ; Hyperplasia ; Immunoglobulin E ; Inflammation* ; Injections, Intraperitoneal ; Intercellular Adhesion Molecule-1 ; Interleukin-6 ; Lovastatin* ; Lung ; Mice ; Mucin 5AC ; Mucus* ; Ovalbumin ; Ovum ; Real-Time Polymerase Chain Reaction