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Objective: To investigate the expressions of heterogeneous nuclear ribonucleoprotein I) (hnRNPD) and epithelial cell adhesion molecule (EpCAM) in the oral squamous cell carcinoma (OSCC) tissue and normal oral mucosa tissue, and to discuss the associations between their expressions and the clinicopathological parameters of the OSCC patients, and to elucidate their effects on the occurrence and development of OSCC. Methods: The tumor tissue of 38 OSCC patients without preoperative radiotherapy and chemotherapy (OSCC group) and the normal oral mucosa tissue of 11 patients with tooth extraction (control group) were selected. Immunohistochemistry staining was used to detect the expression levels of hnRNPI) and EpCAM in the tumor tissue and normal oral mucosa tissue. the expression levels of hnRNPD and EpCAM in tumor tissue of the OSCC patients with different clinicopathological parameters were analyzed∗ and the correlations between the expression levels of hnRNPD and EpCAM in tumor tissue of the OSCC patients were detected by Spearman correlation test. Results: Compared with control group, the expression levels of hnRNPD and EpCAM in the tumor tissue in OSCC group were increased (Z=-4. 936, P= 0.000; Z= 2.780, P=0. 005); there was statistically significant difference in the expression level of EpCAM in tumor tissue of the OSCC patients with different genders and ages (Z=-2.471, P= 0.013; Z=-1.967; P= 0.049), there was no significant difference in the expression level of hnRNPD in tumor tissue of the OSCC patients with different clinicopathological parameters ( P>0. 05). The correlation between the expression levels of hnRNPD and EpCAM was weak (p=0. 227, P=0. 17). Conclusion: The expression levels of hnRNPD and EpCAM in tumor tissue of the OSCC patients are up-regulated, and they promote the occurrence and development of OSCC.
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Epithelial cell adhesion molecule (EpCAM) is a popular target for cancer therapy. In this research, 3 nanobodies with high specificity and endocytosis activity against EpCAM were developed, which provides a basis for the study of immunotoxin based on EpCAM. In our preliminary experiments, we have immunized a camel with EpCAM-Fc antigen and constructed a high-quality phage display library. Seventeen nanobodies with different complementarity determining region (CDR) 3 sequences have been screened after 3 rounds of biopanning by phage display technology. The animal procedures were approved by the Institutional Animal Care and Use Committee (IACUC) of Fudan University School of Pharmacy. After purification, 7 nanobodies showed high cell binding activity by fluorescent activated cell sorting (FACS) identification. Furthermore, 3 nanobodies presented high endocytosis activity based on FACS and laser confocal microscopy, which also showed high affinity to EpCAM measured by ForteBio. According to this study, we aimed to provide a novel alternative approach to the EpCAM-targeted therapy and to provide guidance for the study of nanobody based immunotoxins for other targets.
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Objective To investigate the methods of screening specific aptamers for (EpCAM)Gpositive prostate cancer (PCa)cells by cellGSELEX technique.Methods A random DNA library was designed to screen EpCAMGspecific DNA aptamers from human prostate cancer cells expressing EpCAM molecule by cellGSELEX technique.After 12 rounds of in vitro screening,DNA products were cloned and sequenced.Flow cytometry and cellular immunofluorescence were used to detect the specific binding ability of aptamers to target cells.Results Two aptamers of Ep1 and Ep2 were selected.Both of them could specifically bind to EpCAMGpositive cancer cells LNCap,PCG3 ,DU1 45 , and HEK293T cells transfected with target molecule.The binding rates of Ep1 were 61.0%,74.3%,5 9.1% and 60.3%.The binding rates of Ep2 were 65.1%,77.8%,54.2% and 58.3%.Neither of them could bind to HEK293T cells transfected with empty vector with the binding rate of 5.4% in Ep1 and 3.3% in Ep2,respectively.Flow cytometry analysis and confocal images indicated that the EpCAM aptamers could specifically recognize human PCa cells expressing EpCAM,but could not bind to EpCAMGnegative cells.Conclusion EpCAM aptamers derived from cellGSELEX technology can recognize and bind to EpCAMGpositive PCa cells specifically,which may provide new ideas for the specific diagnosis and targeted therapy of prostate cancer,and lay an experimental basis for the other specific diagnosis and treatment schemes of malignant tumors.
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Objective@#To explore the thermal damage to epithelial cell adhesion molecule(EpCAM)-positive tumor cells by novel aptamer-guided magnetic nanoparticles(AptNPs).@*Methods@#EpCAM aptamer SYL3C was connected to NPs via biotin-streptavidin reaction. The diameter of AptNPs were characterized by Dynamic Light Scattering(DLS). The binding feature of the aptamer to EpCAM-positive tumor cells was evaluated by Prussian blue dyeing. Thermal damage under alternative magnetic field was measured bylactate dehydrogenase (LDH). The apoptosis of EpCAM-positive tumor cells was detected by acridine orange/ethidium bromide (AO/EB) double staining.@*Results@#The average size of AptNPs was 282 nm. Flow cytometry and Prussian blue dyeing showed that AptNPs exhibited strong binding to the EpCAM-positive tumor cells but not to the EpCAM-negative tumor cells. Moreover, when incubated with 1.5×108 AptNPs under alternative electromagnetic fieldfor 5 hours, the viability of EpCAM-positive HCT116 cells and A549 cells was 28.9% and 54.4%, respectively, significantly lower than 76.7% of EpCAM-negative HepG2 cells (P<0.05).@*Conclusions@#AptNPs can improve the thermal damage to EpCAM-positive tumor cells, and may have potential utility in the development of tumor targeted therapy.
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Objective: To investigate the colorectal tumor formation and metastasis of circulating tumor cells in nude mice. Methods: Human colorectal cancer HCT116-luc cells labeled with luciferase were implanted between the mucosa and the base of the cecum wall in nude mice. The circulating tumor cells in peripheral blood in nude mice were screened by epithelial cell adhesion molecule (EpCAM) immunomagnetic beads. The EpCAM-positive (EpCAM+) cells and the EpCAM-negative (EpCAM) cells were injected into the tail vein of the nude mice, respectively, then the tumor formation and the metastases in lung and liver in nude mice were observed. Results: The orthotopic transplanted tumor model of colorectal cancer in nude mice was successfully constructed. The tumor formation rate of EpCAM+circulating tumor cells in nude mice was 90%, the metastasis rates in lung and liver were 60% and 40%, respectively, and the metastasis rate in both liver and lung was 35%. There was no tumor formation of EpCAM circulating tumor cells in nude mice and also no metastasis occurred. The tumor formation rate and the metastasis rate between EpCAM+ and EpCAM groups had statistical differences (all P < 0.01). Conclusion: The tumor formation of EpCAM+ circulating tumor cells in nude mice is good, and the tumor prones to metastasis.
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Objective To study the damage effect of oxidized low density lipoprotein (OX-LDL)on the endothelial cells and its influence in the expressions of Toll like receptor 4 (TLR-4)and epithelial cell adhesion molecule (EpCAM).Methods The human endothelial cells ECV-304 were cultured in vitro and treated by different concentrations of OX-LDL for 24 h,and the cell vitality was measured by Taipan blue rejection test.The reactive oxygen species (ROS)level,mitochondrial membrane potential (MMP),cell cycle and apoptosis were detected by flow cytometry (FCM).The expression of TLR-4 and EpCAM were also analyzed by FCM.Results Compared with control group,the ROS levels in ECV-304 cells in 25,50,100,and 200 mg·L-1 OX-LDL were increased, but the cell vitalities and MMP were decreased (P < 0.05).Compared with control group,the percentages of S phase cells in 25 and 50 mg·L-1 OX-LDL groups were increased,the G0 G1 phase cells were slightly reduced,and the SubG1 phase cells (apoptotic cells)were increased with the increasing of OX-LDL concentration.Compared with control group,the TLR-4 and EpCAM expression levels in ECV-304 cells in 50 mg·L-1 OX-LDL group at 24 h were significantly increased (P < 0.05 ). Conclusion OX-LDL could increase the ROS level and induce the apoptosis in vascular endothelial cells,while increase the cell vitality and MMP.During the damage process,the expresion levels of TLR-4 and EpCAM are up-regulated.
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Tufting enteropathy (TE), also known as intestinal epithelial dysplasia (IED), is a rare congenital enteropathy related to an earlyonset of severe intractable diarrhea due to specific abnormalities of the intestinal epithelium and mutations of the EpCAM gene. TE is characterized by clinical and histological heterogeneity, such as with low or without mononuclear cell infiltration of the lamina propria, and abnormalities of basement membrane. TE can be associated with malformations, other epithelial diseases, or to abnormal enterocytes development and/or differentiation. The authors report a case of a Brazilian child with TE associated with c.556-14A>G mutation in the EpCAM gene (NM_002354.2)...
Enteropatia com formação de tufos epiteliais (ETE), também conhecida como displasia epitelial intestinal (DEI), é uma rara enteropatia congênita relacionada com um início precoce de diarreia intratável grave devido a anormalidades específicas do epitélio intestinal e mutações do gene EpCAM. ETE caracteriza-se por uma heterogeneidade clínica e histológica, como ausência ou leve infiltrado de células mononucleares na lâmina própria e anormalidades de membrana basal. Pode ser associada a malformações, outras doenças epiteliais ou anormalidades no desenvolvimento/na diferenciação dos enterócitos. Os autores relatam um caso de ETE, em uma criança brasileira, associada à mutação c.556-14A> g do gene EPCAM (NM_002354.2)...
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Humans , Female , Child , Epithelial Cells/pathology , Intestinal Diseases/genetics , Cell Adhesion Molecules/genetics , Diarrhea, Infantile , Intestinal Mucosa/pathologyABSTRACT
OBJECTIVE: Epithelial cell adhesion molecule (EpCAM) has experienced a renaissance lately as a binding site for targeted therapy as well as a prognostic marker in epithelial malignancies. Aim of this study was to study EpCAM as a potential prognostic marker in epithelial ovarian cancer (EOC). METHODS: EpCAM expression was assessed by immunohistochemistry on paraffin-embedded primary EOC-tissue samples. EpCAM overexpression was defined as an expression of EpCAM of 76% to 100%. Tissue samples and clinical data were systematically collected within the international and multicenter "Tumorbank Ovarian Cancer" network. RESULTS: Seventy-four patients, diagnosed with EOC between 1994 and 2009, were included in the study (median age, 56 years; range, 31 to 86 years). The majority of the patients (81.1%) presented with an advanced stage International Federation of Gynecology and Obstetrics (FIGO) III/IV disease. Histology was of the serous type in 41 patients (55.4%), endometrioid in 19 (25.6%), and mucinous in 14 (19%). EpCAM was overexpressed in 87.7%. Serous tumors overexpressed EpCAM significantly more often than mucinous tumors (87.8% vs. 78.6%, p=0.045); while no significant difference was noted between the other histological subgroups. EpCAM overexpression was significantly associated with a better progression free survival and higher response rates to platinum based chemotherapy (p=0.040 and p=0.048, respectively). EpCAM was identified as an independent prognostic marker for overall survival (p=0.022). CONCLUSION: Our data indicate a significant association of EpCAM overexpression with a more favorable survival in EOC-patients. Serous cancers showed a significant EpCAM overexpression compared to mucinous types. Larger multicenter analyses are warranted to confirm these findings.
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Middle Aged , Antigens, Neoplasm/metabolism , Antineoplastic Agents/therapeutic use , Carboplatin/therapeutic use , Cell Adhesion Molecules/metabolism , Kaplan-Meier Estimate , Neoplasm Proteins/metabolism , Neoplasm Staging , Neoplasms, Glandular and Epithelial/diagnosis , Organoplatinum Compounds/therapeutic use , Ovarian Neoplasms/diagnosis , Paclitaxel/therapeutic use , Prognosis , Tissue Banks , Treatment Outcome , Biomarkers, Tumor/metabolismABSTRACT
Objective To prepare and identify the monoclonal antibody (mAb) specific to epithelial cell adhesion molecule (EpCAM) and explore the function of the mAb.Methods The EpCAM antigen expressed by the prokaryotic expression systems was used to immunize the BALB/c mice,and then the splenic cells from the mice were fused with the Sp2/0 cells to produce hybridomas secreting specific mAb.The positive clones were screened by the ELISA.The western blot analysis was used to identify the reactivity of the mAb to the antigen.Then the immunohistochemistry staining was used to detect EpCAM expression in the 3 primary colorectal carcinoma tissues.Results Three mAb specific to EpCAM were obtained by ELISA tests.Western blot results indicated that these three kinds of antibodies could react to the EpCAM antigen,but no response to the GST tag.Immunohistochemical staining results identified that these mAb could give positive signals to the primary colorectal carcinoma tissues from patients.Conclusion Three mAb specific to EpCAM are obtained and identified,which contributes to the diagnosis and therapy of the carcinoma in the future.
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Objective: To investigate the expressions of tumor stem cell markers EpCAM (epithelial cell adhesion molecule) and CD133 proteins in human primary hepatocellular carcinoma, and to explore their relationships with clinicopathological features and the prognosis. Methods: The expressions of EpCAM and CD133 in liver cancer tissues from 70 patients with primary hepatocellular carcinoma were detected by immunohistochemistry. The survival analysis was performed, and the correlations of EpCAM and CD133 with the clinicopathological features and the prognosis were analyzed using SPSS (Statistical Package for the Social Sciences) software package 17.0. Results: The positive rates of CD133 and EpCAM proteins were both significantly higher in liver cancer tissues than those in paracancerous tissues (P < 0.01). Aberrant expression of CD133 was observed in the paracancerous tissues. The positive expression of CD133 was positively correlated with vascular invasion in liver cancer tissues (P < 0.05). The expression level of EpCAM in poorly differentiated liver cancer with vascular invasion in patients with preoperative AFP level over 400 ng/mL was significantly higher (P < 0.05). There was a positive correlation between EpCAM expression and CD133 expression (P < 0.01). The survival of patients with strongly positive expression of CD133 (or EpCAM) was inferior to that of patients with moderately-weakly positive expression of CD133 (or EpCAM). In the subgroup of preoperative AFP level over 400 ng/mL, the survival of patients with moderately-strongly positive expression of EpCAM was inferior to that of patients with weakly positive expression of EpCAM. The factors of the number of tumors, preoperative AFP level, tumor capsule and CD133 expression were independent predictors of prognosis of primary hepatocellular carcinoma. Conclusion: CD133 and EpCAM may be associated with the progression of primary hepatocellular carcinoma, and their high expression may indicate poor prognosis, thus it is suggested EpCAM (+) AFP (+) primary hepatocellular carcinoma subtype has features of high-grade malignancy. Copyright © 2012 by TUMOR.
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Tufting enteropathy is a rare autosomal recessive disorder presenting with early-onset severe intractable diarrhea. The epithelial cell adhesion molecule gene (EpCAM) has recently been identified as the gene responsible for tufting enteropathy. Based on histology, a diagnosis of tufting enteropathy was made in two Korean siblings. They developed chronic diarrhea and failure to thrive. They had a broad nasal bridge and micrognathia. Duodenal and colonic biopsies showed villous atrophy, disorganization of surface enterocytes, and focal crowding resembling tufts. Protracted diarrhea continued and so cyclic parenteral nutrition was supplied. The sister had juvenile rheumatoid arthritis. Mutation analysis of EpCAM identified two compound heterozygous mutations in these siblings: 1) a donor splicing site mutation in intron 5 (c.491+1G>A) and 2) a novel nonsense mutation in exon 3 (c.316A>T, Lys106X). Analysis of EpCAM will be useful for genetic counseling and prenatal diagnosis of tufting enteropathy.