Analysis of Significant Genes and Pathways in Esophageal Cancer Based on Gene Expression Omnibus Database / 中国医学科学杂志(英文版)

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.

Terapia com células CAR-T: reprogramação celular para o combate de neoplasias malignas / CAR-T cell therapy: cell reprogramming to combat malignant neoplasms

As células CAR-T são linfócitos geneticamente modificados para reconhecerem um espectro amplo de antígenos de superfície celulares. Além disso, atacam células tumorais malignas, que expressam esses antígenos, por meio da ativação da coestimulação citoplasmática, secreção de citocinas, citólise de células tumorais e proliferação de células T. O objetivo desse estudo é abordar a imunoterapia com células CAR-T, a fim de explicar seu conceito, processo de fabricação e papel no tratamento de neoplasias hematológicas e tumores sólidos. Foi realizada uma revisão através do portal PubMed, utilizando como descritores: "car-t cell therapy" e "neoplasms", determinados com base nos "Descritores em Ciências da Saúde". Foram obtidos, inicialmente, 10 artigos, os quais foram lidos integralmente para a confecção dessa revisão. Além disso, foram adicionados 3 ensaios clínicos atualizados sobre o tema. Na terapia com células CAR-T, as células T são coletadas do paciente, geneticamente modificadas para incluir receptores de antígeno específicos e, posteriormente, expandidas em laboratórios e transfundidas de volta para o paciente. Assim, esses receptores podem reconhecer células tumorais que expressam um antígeno associado a um tumor. A terapia com células CAR-T é mais conhecida por seu papel no tratamento de malignidades hematológicas de células B, sendo a proteína CD19 o alvo antigênico mais bem estudado até o momento. Entretanto, estudos estão sendo feitos para verificar a eficácia desse tratamento, também, em tumores sólidos. Portanto, apesar de inicialmente ser indicada apenas para um grupo seleto de pessoas, essa terapia tem demonstrado grande potencial para atuar em um espectro maior de pacientes.

The CAR-T cells are lymphocytes genetically modified to recognize a broader spectrum of cell surface antigens. In addition, they attack malignant tumor cells, which express these antigens, by activating cytoplasmic co-stimulation, cytokine secretion, tumor cell cytolysis and T cell proliferation. The aim of this study is to address immunotherapy with CAR-T cells, in order to explain its concept, manufacturing process and role in the treatment of hematological neoplasms and solid tumors. This is a literature review conducted through the PubMed portal, that uses the terms "car-t cell therapy" and "neoplasms" as descriptors, determined based on the DeCS (Descritores em Ciências da Saúde). To prepare this review, initially 10 articles were found and read in full. In addition, 3 updated clinical trials on the subject were added. For CAR-T cell therapy, T cells are collected from the patient, genetically modified to include specific antigen receptors, and later expanded in laboratories and transfused back to the patient. Thus, these receptors can recognize tumor cells that express a tumor-associated antigen. CAR-T cell therapy is best known for its role in the treatment of B cell hematological malignancies, with the CD19 protein being the most studied antigenic target to date. However, studies are being conducted to verify the effectiveness of this treatment, also, in solid tumors. Therefore, despite being formulated only for a selected group of patients, this therapy has great potential to act on a broader spectrum of patients.

Frequency of Variants in DNA-Repair Genes in a Southwest Colombian Population / Frecuencia de las variantes en genes de reparación del ADN en una población del suroccidente de Colombia

Objective To describe the frequency of mutations in DNA-repair genes in a southwestern Colombian population. Methods We have designed an observational study, including 162 people from all ages from southwest Colombia. We have extracted and collected their DNA in filters. We have immersed the DNA in a phosphate buffer along with DNeasy package (Thermo Fisher Scientific, Waltham, MA, USA). The preparation process was with the TruSeq Exome Library Prep (Illumina, Inc. San Diego, CA, USA), then the obtained libraries were normalized with TruSeq Rapid Exome (Illumina, Inc. San Diego, CA, USA). We sequenced the full exome and identified the variants associated with 12 genes (ataxia telangiectasia mutated [ATM], BRCA1 DNA repair associated [BRCA1], BRCA2 DNA repair associated [BRCA2], checkpoint kinase 2 [CHEK2], epithelial cell adhesion molecule [EPCAM], homeobox protein Hox-B13 [HOXB13], mutS homolog 1, 2 and 6 [MLH1, MSH2, MSH6], nibrin [NBN], PMS1 homolog 2, mismatch repair system component [PMS2], and tumor protein p53 [TP53]). Descriptive statistics were performed with the R software (The R Foundation for Statistical Computing, Vienna, Austria). Results A total of 7,315,466 pieces of data were sequenced in this population. The most frequently mutated genes were ATM (1,221 pieces of data; 13.2%), BRCA1 (1,178 pieces of data; 12.8%), BRCA2 (1,484 pieces of data; 16.12%), and NBN (965 pieces of data; 10.42%). The most common single nucleotide polymorphisms (SNPs) in these 12 genes were the following: BRCA2 (rs169547, rs206075, rs206076); ATM (rs659243, rs228589); TP53 (rs1625895, rs1042522, rs1642785); PMS2 (rs2228006, rs1805319); NBN (rs709816); and MSH6 (rs3136367) Conclusion The BRCA2, ATM, BRCA1 and NBN DNA-repair genes were the most frequently mutated in this southwestern Colombian Population

Objetivo Describir la frecuencia de las mutaciones en los genes de reparación del ADN en una población del suroccidente de Colombia. Métodos Diseñamos un estudio observacional que incluyó a 162 personas del suroccidente de Colombia de todas las edades. Hemos extraído y recogido el ADN en filtros. Los sumergimos en tampón fosfato junto con el paquete DNeasy (Thermo Fisher Scientific, Waltham, MA, EEUU). El proceso de preparación fue realizado con TruSeq Exome Library Prep (Illumina, Inc. San Diego, CA, EEUU); luego, las bibliotecas obtenidas se normalizaron con TruSeq Rapid Exome (Illumina, Inc. San Diego, CA, USA). Secuenciamos el exoma completo e identificamos las variantes asociadas a doce genes (ataxia telangiectasia mutated [ATM], BRCA1 DNA repair associated [BRCA1], BRCA2 DNA repair associated [BRCA2], checkpoint kinase 2 [CHEK2], epithelial cell adhesion molecule [EPCAM], homeobox protein Hox-B13 [HOXB13], mutS homolog 1, 2 and 6 [MLH1, MSH2, MSH6], nibrin [NBN], PMS1 homolog 2, mismatch repair system component [PMS2], y tumor protein p53 [TP53]). La estadística descriptiva se realizó en el programa R (The R Foundation for Statistical Computing, Viena, Austria). Resultados Un total de 7.315.466 datos fueron secuenciados en esta población. Los genes más frecuentemente mutados fueron el ATM, con 1.221 datos (13,2%), el BRCA1, con 1.178 datos (12,8%), el BRCA2, con 1.484 datos (16,12%) y el NBN, con 965 datos (10,42%). Los polimorfismos de un solo nucleótido (PSN) más comunes en estos 12 genes fueron los siguientes: BRCA2 (rs169547, rs206075, rs206076); ATM (rs659243, rs228589); TP53 (rs1625895, rs1042522, rs1642785); PMS2 (rs2228006, rs1805319); NBN (rs709816) y MSH6 (rs3136367) Conclusión Los genes de reparación de ADN BRCA2, ATM, BRCA1 NBN fueron los más frecuentemente mutados en esta población del suroccidente de Colombia.

Cancer Stem Cells are Depolarized Relative to Normal Stem Cells Derived from Human Livers

ABSTRACT Introduction and aim. The inability to distinguish cancer (CSCs) from normal stem cells (NSCs) has hindered attempts to identify safer, more effective therapies for hepatocellular carcinoma (HCC). The aim of this study was to document and compare cell membrane potential differences (PDs) of CSCs and NSCs derived from human HCC and healthy livers respectively and determine whether altered GABAergic innervation could explain the differences. Material and methods. Epithelial cell adhesion molecule (EpCAM) positive stem cells were isolated from human liver tissues by magnetic bead separations. Cellular PDs were recorded by microelectrode impalement of freshly isolated cells. GABAA receptor subunit expression was documented by reverse transcriptase polymerase chain reaction (RT-PCR) and immunofluorescence. Results. CSCs were significantly depolarized (-7.0 ± 1.3 mV) relative to NSCs (-23.0 ± 1.4 mV, p < 0.01). The depolarized state was associated with different GABAA receptor subunit expression profiles wherein phasic transmission, represented by GAGAA α3 subunit expression, was prevalent in CSCs while tonic transmission, represented by GABAA α6 subunit expression, prevailed in NSCs. In addition, GABAA subunits α3, β3, γ3 and δ were strongly expressed in CSCs while GABAA π expression was dominant in NSCs. CSCs and NSCs responded similarly to GABAA receptor agonists (ΔPD: 12.5 ± 1.2 mV and 11.0 ± 3.5 mV respectively). Conclusion. The results of this study indicate that CSCs are significantly depolarized relative to NSCs and these differences are associated with differences in GABAA receptor subunit expression. Together they provide new insights into the pathogenesis and possible treatment of human HCC.

Expressions of epithelial cell adhesion molecule, vimentin and N-cadherin in molecular subtypes of breast cancer and the correlation among them / 中南大学学报(医学版)

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.

Expression of epithelial cell adhesion molecule and molecular subtypes and prognosis of breast cancer / 中南大学学报(医学版)

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.

Comparing the results of pathologic diagnosis of mesothelioma between Chinese and Japanese experts / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>The 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.</p><p><b>RESULTS</b>Twenty-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.</p><p><b>CONCLUSION</b>The 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.</p>

Expression of the epithelial cell adhesion molecule in prostate cancer and its clinical significance / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the expression of the epithelial cell adhesion molecule (EpCAM) in prostate cancer (PCa) and its clinical significance.</p><p><b>METHODS</b>We 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.</p><p><b>RESULTS</b>The 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).</p><p><b>CONCLUSION</b>The 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.</p>

EPCAM-positive normal hepatic progenitor cells transformation into liver stem cells and HBx-mediated effects on stability in adult mouse / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>Hepatic 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.</p><p><b>RESULTS</b>Cell 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.</p><p><b>CONCLUSION</b>EPCAM-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.</p>

Expression of EpCAM and E-cadherin in papillary thyroid carcinoma and its clinicopathologic significance / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the expression of EpCAM and E-cadherin in papillary thyroid carcinoma and to analyze its correlation with various clinicopathologic parameters.</p><p><b>METHODS</b>Immunohistochemical 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.</p><p><b>RESULTS</b>In 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).</p><p><b>CONCLUSIONS</b>E-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.</p>

Detection of circulating tumor cells in patients with hepatocellular carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the detection efficiency of circulating tumor cells (CTCs) in patients with hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>Immunomagnetic negative enrichment by nanometer magnetic beads and label-free capture with Captor(TM) system were used to isolate and enrich CTCs from peripheral blood of HCC patients, and epithelial and HCC markers were applied to identify CTCs by immunofluorescence staining. CTCs were detected in 50 HCC patients before and after hepatectomy to test the method for isolation, enrichment and identification. The dynamic changes of pre- and post-operative CTCs' numbers were compared. The clinical data were analyzed using SPSS 19.0 software.</p><p><b>RESULTS</b>Negative enrichment methods by nanometer magnetic beads and label-free capture using Captor(TM) system were both suitable for CTCs isolation and enrichment in HCC patients. The positive detection rate of CTCs in HCC patients via negative enrichment was 96.0% (48/50), the preoperative median number of CTCs was 16 per 7.5 ml blood, and the postoperative median number was 17 per 7.5 ml blood.</p><p><b>CONCLUSIONS</b>Both negative enrichment and Captor(TM) system are suitable for isolation and enrichment of CTCs in HCC patients. There is a significant difference in the numbers of CTCs before and after operation, and dynamic detection of CTCs will provide helpful prognostic information for HCC patients in clinics.</p>

Cell morphology in the dormancy and proliferation stage of colorectal cancer stem cells / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>The 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.</p><p><b>RESULTS</b>The 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.</p><p><b>CONCLUSIONS</b>Cancer 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.</p>

Expression of CD90/EpCAM/CD24 in hepatocellular carcinoma cell lines at various stages of differentiation / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To confirm the malignant phenotype of hepatocarcinoma cell (HCC) lines at various stages of differentiation (MHCC97L, MHCC97H and HCCLM3) and to explore their expression levels of cancer stem cell (CSC) markers.</p><p><b>METHODS</b>The invasive and proliferative properties of each HCC line were assessed by transwell assay and the Cell Counting Kit-8 (CCK-8) colorimetric assay. Sensitivity to chemotherapy was assessed by treatment with oxaliplatin and determination of the half inhibitory concentration (IC50). The expression of CD90, EpCAM and CD24 was measured by flow cytometry.</p><p><b>RESULTS</b>The number of cells that migrated through the invasion assay membrane were significantly different between the three HCC lines: HCCLM3 (30.57 +/- 8.95) more than MHCC97H (21.33 +/- 4.17) more than HCC97L (9.33 +/- 3.85), P less than 0.01. The IC50 was significantly different between the three HCC lines: HCCLM3 (36.57 +/- 6.95) mumol/L more than MHCC97H (26.35+/-3.88) mumol/L more than MHCC97L (17.68 +/- 3.25) mumol/L. The CSC marker with the highest expression on all three HCC lines was CD90 (HCCLM3: 0.92% +/- 0.21%, MHCC97H: 1.98% +/- 0.23%, and MHCC97L: 2.55% +/- 0.34%), followed by EpCAM (2.11% +/- 0.32%, 3.23% +/- 0.18%, and 4.38% +/-0.49%, respectively), and CD24 as the lowest (0.68% +/- 0.37%, 1.22% +/- 0.26%, and 1.36% +/- 0.24%, respectively).</p><p><b>CONCLUSION</b>Higher expression of CSC markers on HCC lines is associated with a stronger invasive ability and higher sensitivity to chemotherapy.</p>

Effect of epithelial cell adhesion molecule on metastasis in hypopharyngeal carcinoma FaDu cells / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of knockdown of EpCAM by siRNA on invasion, migration, and colony abilities in hypopharyngeal carcinoma FaDu cells.</p><p><b>METHODS</b>A siRNA against EpCAM was employed to inhibit the expression of EpCAM in FaDu cells. Measurements included the Transwell assay for invasion and migration, plate colony formation assay for cell colony ability, Western blot assay for EpCAM, E-cadherin, and β-catenin expressions in total protein, cytoplasm, and cytoskeleton, respectively.</p><p><b>RESULTS</b>mRNA and protein expressions of EpCAM were suppressed significantly in FaDu cells transfected by EpCAM siRNA (t = 6.46, P < 0.05; t = 10.25, P < 0.05) . Transwell assay showed in transwell assay, the average invasive cells in EpCAM siRNA cells (26.33 ± 3.71) was less than that in FaDu cells (61.47 ± 6.70; t = 7.95, P < 0.05)and control cells (54.13 ± 6.51; t = 6.42, P < 0.05); the average number of migration cells in EpCAM siRNA cells (79.87 ± 8.44) was lower than that in FaDu (167.53 ± 11.49; t = 10.90, P < 0.05) cells and control cells (162.13 ± 13.45; t = 8.97, P < 0.05). In plate colony formation assay, the average colony number of EpCAM siRNA cells was (78.00 ± 5.57), which was less than that of FaDu cells(177.30 ± 16.50; t = 9.78, P < 0.05) and control cells (173.67 ± 13.50; t = 11.35, P < 0.05). Western blot assays showed, silencing of EpCAM increased the expressions of E-cadherin (t = 4.58, P = 0.01) and β-catenin (t = 3.76, P = 0.02) in cytoskeleton, and decreased the expressions of E-cadherin (t = 6.60, P < 0.05) and β-catenin (t = 8.20, P < 0.05) in cytoplasm.</p><p><b>CONCLUSIONS</b>The knockdown of EpCAM inhibits the invasion, migration, and colony formation abilities of FaDu cells, which is probably related to the regulation of E-cadherin and β-catenin in cytoplasm and cytoskeleton, and EpCAM may be a promising gene therapy target for hypopharyngeal carcinoma.</p>

A case of primary liver carcinosarcoma: CT findings / 癌症

Primary liver carcinosarcoma is extremely rare. We report a case of liver carcinosarcoma in a 58-year-old man, which was identified by pathologic and immunohistochemical (IHC) examinations. Plain CT scans showed a hypodense mass in the right liver lobe adjacent to the gallbladder fossa. The triple-phase contrast CT scans showed a mixed density mass with inhomogeneous enhancement. Multiple cystic nodules with irregular rim enhancement of the margin located in the tumor. The gallbladder wall and transverse colon were involved. CT presentations of liver carcinosarcoma were unspecific and the pre-operative diagnosis is difficult.

Expressions of Ep-CAM and cyclin D1 and their clinical significance in gallbladder carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the expressions of Ep-CAM and cyclin D1 and their correlation with the clinicopathological factors and prognosis in gallbladder carcinoma.</p><p><b>METHODS</b>Ep-CAM and cyclin D1 expressions were detected by PV6000 immunohistochemical staining in 60 gallbladder carcinoma and 15 para-cancerous mucosa specimens.</p><p><b>RESULTS</b>Ep-CAM and cyclin D1 expressions were detected in 56.7% and 48.3% of the cancer tissues, respectively, significantly higher than those in the normal mucosa (P < 0.05). Ep-CAM expression was not correlated with clinicopathological data, however cyclin D1 expression was correlated with pathological differentiation and necrosis (P < 0.05). Survival analysis showed that patients with positive Ep-CAM or cyclin D1 expression had a shorter survival time than that in the patients without (P < 0.05). Moreover, Ep-CAM and cyclin D1 expressions were positively correlated with each other (r = 0.307, P = 0.017).</p><p><b>CONCLUSION</b>Ep-CAM or cyclin D1 expression is a unfavorable prognostic factor in gallbladder carcinoma.</p>

Current advances in gene diagnosis and therapy of gelatinous drop-like corneal dystrophy / 浙江大学学报·医学版

Gelatinous drop-like corneal dystrophy (GDLD) is an autosomal recessive hereditary disease, which may result in bilateral loss of vision. The gene responsible for GDLD, M1S1 is mapped on the short arm of chromosome 1 (1p), but the possible etiology of this disease remains unclear. Corneal transplantation is the only treatment for visual rehabilitation. The detection of the mutations of the M1S1 gene and the possible etiological involvement of the amyloid deposits are discussed. The current literatures are extensively reviewed in this article.