Cadherin 1 Research Articles

Long noncoding RNA NSCLCAT1 increases non-small cell lung cancer cell invasion and migration through the Hippo signaling pathway by interacting with CDH1.

Metastatic growth is the leading cause of cancer-related death in non-small cell lung cancer (NSCLC). Metastasis is believed to be initiated by an increase in cell motility mediated by the loss of cell-cell adhesion because of the suppression of E-cadherin [encoded by cadherin 1 ( CDH1)]. However, very little is known about the molecular mechanism of CDH1 regulation. Therefore, we hypothesized that non-small cell lung cancer-associated transcript-1 (NSCLCAT1) suppresses functional CDH1 and mediates the Hippo signaling pathway, resulting in increased cell migration and invasion, and reduced apoptosis. Initially, microarray profiling and target prediction programs were employed to identify whether NSCLCAT1 targets CDH1. Next, quantitative PCR was used to determine the expression pattern of NSCLCAT1 in 114 specimens. The biologic functions of NSCLCAT1 in NSCLC were assessed through the up-regulation and down-regulation of the levels of endogenous NSCLCAT1 with the use of NSCLCAT1 vector or small interfering RNA against NSCLCAT1 in NSCLC cells. Furthermore, the Hippo signaling pathway in NSCLC cells was blocked by applying the verteporfin treatment to have a better understanding on the pivotal role of the Hippo signaling pathway in NSCLC. Microarray expression profiles of long noncoding RNAs, GSE19804 and GSE27262), revealed that NSCLCAT1 was up-regulated in NSCLC. Among patients with NSCLC, we determined that the NSCLCAT1 was robustly induced, whereas CDH1 was suppressed. The luciferase activity determination identified CDH1 as a NSCLCAT1 target. NSCLCAT1 was found to increase cell viability, migration, and invasion and to reduce apoptosis in NSCLC cells. The results from the quantitative PCR and Western blot analysis revealed that NSCLCAT1 modulated the Hippo signaling pathway. Furthermore, the inhibition of the Hippo signaling pathway by verteporfin treatment led to the loss of the effect of NSCLCAT1 on NSCLC cells. In summary, our findings suggested that NSCLCAT1 potentially has a role in NSCLC and NSCLCAT1-mediated regulation of the Hippo signaling pathway through the transcriptional repression of CDH1; therefore, the functional suppression or inhibition of NSCLCAT1 could be used as a novel therapeutic pathway in the control of aggressive and metastatic NSCLC.-Zhao, W., Zhang, L.-N., Wang, X.-L., Zhang, J., Yu, H.-X. Long noncoding RNA NSCLCAT1 increases non-small cell lung cancer cell invasion and migration through the Hippo signaling pathway by interacting with CDH1.

Prediction of key genes and pathways involved in trastuzumab-resistant gastric cancer

BackgroundTrastuzumab has been prevailingly accepted as a beneficial treatment for gastric cancer (GC) by targeting human epidermal growth factor receptor 2 (HER2)-positive. However, the therapeutic resistance of trastuzumab remains a major obstacle, restricting the therapeutic efficacy. Therefore, identifying potential key genes and pathways is crucial to maximize the overall clinical benefits.MethodsThe gene expression profile GSE77346 was retrieved to identify the differentially expressed genes (DEGs) associated with the trastuzumab resistance in GC. Next, the DEGs were annotated by the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The DEGs-coded protein-protein interaction (PPI) networks and the prognostic values of the 20 hub genes were determined. Correlation of the hub genes were analyzed in The Cancer Genome Atlas. The prognostic values of hub genes were further validated by Kaplan-Meier (KM) plotter.ResultsA total of 849 DEGs were identified, with 374 in upregulation and 475 in downregulation. Epithelium development was the most significantly enriched term in biological processes while membrane-bounded vesicle was in cellular compartments and cell adhesion molecular binding was in molecular functions. Pathways in cancer and ECM-receptor interaction were the most significantly enriched for all DEGs. Among the PPI networks, 20 hub genes were defined, including CD44 molecule (CD44), HER-2, and cadherin 1 (CDH1). Six hub genes were associated with favorable OS while eight were associated with poor OS. Mechanistically, 2′-5′-oligoadenylate synthetase 1, 3 (OAS1, OAS3) and CDH1 featured high degrees and strong correlations with other hub genes.ConclusionsThis bioinformatics analysis identified key genes and pathways for potential targets and survival predictors for trastuzumab treatment in GC.

Candidate genes and microRNAs for glioma pathogenesis and prognosis based on gene expression profiles.

Glioma is the most common malignant brain tumor, and the incidence of glioma demonstrates an upward trend. It is vital to elucidate the pathogenesis of glioma and seek effective therapies. The aim of the present study was to identify the potential gene markers associated with glioma based on GSE31262 gene expression profiles, and to explore the underlying mechanism of glioma progression by analyzing the gene markers. The microarray dataset GSE31262 was downloaded and neural stem cell samples (control group) and glioma samples (glioma group) were analyzed to identify the differentially expressed genes (DEGs) between the two groups. Gene Ontology functional and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed using DAVID software. Subsequently, a protein-protein interaction (PPI) network was constructed and important modules were extracted from this network. Additionally, the miRNA-target regulatory network was established. In total, 1377 DEGs with P<0.01 and |log2 fold change| ≥2 were identified between the control and glioma groups. The DEGs that were upregulated in glioma samples compared with controls were primarily associated with functions such as the M phase and cell cycle pathway, while the downregulated genes were associated with functions such as nerve impulse and the axon guidance pathway. The results also indicated that certain DEGs, including cyclin-dependent kinase 1 (CDK1) and cadherin 1 (CDH1), had important roles in the PPI network. The MCODE tool in Cytoscape software was used to identify upregulated and downregulated modules in the PPI network, and 5 upregulated and 2 downregulated modules were extracted. Furthermore, the WebGestal online tool was used to identify potential interactions of the upregulated and downregulated genes with microRNAs (miRNA/miR), and miR-135A/B and its two targets, discs large MAGUK scaffold protein 2 and forkhead box O1 (FOXO1), had the highest number of connections in the miRNA-target regulatory network. In addition, cell division cycle 20 and FOXO1 were confirmed to be upregulated in U87 glioma cells compared with normal human astrocytes (HA1800) by reverse transcription-quantitative polymerase chain reaction. In conclusion, M phase function and the axon guidance pathway may be vital for glioma progression. In addition, CDK1 and CDH1 may be associated with the process of glioma. Furthermore, miR-135A/B, and the target FOXO1, may be potential therapy targets for glioma treatment.

A novel de novo CDH1 germline variant aids in the classification of carboxy-terminal E-cadherin alterations predicted to escape nonsense-mediated mRNA decay.

Most truncating cadherin 1 (CDH1) pathogenic alterations confer an elevated lifetime risk of diffuse gastric cancer (DGC) and lobular breast cancer (LBC). However, transcripts containing carboxy-terminal premature stop codons have been demonstrated to escape the nonsense-mediated mRNA decay pathway, and gastric and breast cancer risks associated with these truncations should be carefully evaluated. A female patient underwent multigene panel testing because of a personal history of invasive LBC diagnosed at age 54, which identified the germline CDH1 nonsense alteration, c.2506G>T (p.Glu836*), in the last exon of the gene. Subsequent parental testing for the alteration was negative and additional short tandem repeat analysis confirmed the familial relationships and the de novo occurrence in the proband. Based on the de novo occurrence, clinical history, and rarity in general population databases, this alteration was classified as a likely pathogenic variant. This is the most carboxy-terminal pathogenic alteration reported to date. Additionally, this alteration contributed to the classification of six other upstream CDH1 carboxy-terminal truncating variants as pathogenic or likely pathogenic. Identifying the most distal pathogenic alteration provides evidence to classify other carboxy-terminal truncating variants as either pathogenic or benign, a fundamental step to offering presymptomatic screening and prophylactic procedures to the appropriate patients.

Microarray screening for key genes and prognosis factors in interferon regulatory factor 1-silenced ovarian cancer SKOV-3 cells

Background: Ovarian cancer is a gynecological neoplastic disease with high mortality rate. Its early detection is difficult because of the lack of specific clinical symptoms. This study aimed to identify key genes and prognosis factors associated with ovarian cancer to provide new information and thus better understanding of ovarian cancer. Methods: Microarray data from the Gene Expression Omnibus (GEO) database (accession number GSE38551) were used for analysis. Differentially expressed genes (DEGs) were screened, and functional enrichment and protein-protein interaction (PPI) network analyses for DEGs were performed. A subnetwork was constructed to gain further information regarding DEGs scored in the PPI network. Finally, we performed survival analysis. Results: In total, 427 DEGs were obtained in interferon regulatory factor 1 ( IRF-1 )-silenced ovarian cancer SKOV-3 cell line samples compared to SKOV-3 samples without IRF-1 silencing. DEGs were mainly enriched in metabolic pathways and systemic lupus erythematosus. Tumor necrosis factor ( TNF ) and cadherin 1 ( CDH1 ; type 1, E-cadherin) were present in had higher degrees than others in both the PPI network and the subnetwork. The subnetwork results presented that CDH1 was enriched in the epithelium morphogenesis and cancer pathways, and TNF was enriched in response to lipids. The Mir-30 family served as a tumor suppressor in ovarian cancer. Survival analysis revealed that CDH1 was associated with ovarian cancer prognosis. Conclusions: TNF and CDH1 play important roles in ovarian cancer: CDH1 is an important prognosis factor for ovarian cancer and may be involved mainly via epithelial morphogenesis and cancer pathways. TNF may be involved via response to lipids.

Bioinformatics and functional analyses of key genes and pathways in human clear cell renal cell carcinoma.

Clear cell renal cell carcinoma (ccRCC) is the most common type of kidney cancer. The present study was conducted to explore the mechanisms and identify the potential target genes for ccRCC using bioinformatics analysis. The microarray data of GSE15641 were screened on Gene-Cloud of Biotechnology Information (GCBI). A total of 32 ccRCC samples and 23 normal kidney samples were used to identify differentially expressed genes (DEGs) between them. Subsequently, the clustering analysis and functional enrichment analysis of these DEGs were performed, followed by protein-protein interaction (PPI) network, and pathway relation network. Additionally, the most significant module based on PPI network was selected, and the genes in the module were identified as hub genes. Furthermore, transcriptional level, translational level and survival analyses of hub genes were performed to verify the results. A total of 805 genes, 403 upregulated and 402 downregulated, were differentially expressed in ccRCC samples compared with normal controls. The subsequent bioinformatics analysis indicated that the small molecule metabolic process and the metabolic pathway were significantly enriched. A total of 7 genes, including membrane metallo-endopeptidase (MME), albumin (ALB), cadherin 1 (CDH1), prominin 1 (ROM1), chemokine (C-X-C motif) ligand 12 (CXCL12), protein tyrosine phosphatase receptor type C (PTPRC) and intercellular adhesion molecule 1 (ICAM1) were identified as hub genes. In brief, the present study indicated that these candidate genes and pathways may aid in deciphering the molecular mechanisms underlying the development of ccRCC, and may be used as therapeutic targets and diagnostic biomarkers of ccRCC.

Fat atypical cadherin 1 regulating invasion by targeting hypoxia inducible factor-1α in high grade glioma

Objective To investigate the action mechanism of fat atypical cadherin 1 (FAT1) regulating invasion by targeting hypoxia inducible factor-1α (HIF-1α) in high grade glioma under hypoxia. Methods The expression levels of FAT1 and HIF-1α in 48 cases of grade Ⅲ-Ⅳ glioma athological tissue samples in our hospital were detected by real-time fluorescent quantitative polymerase chain reaction (FQ-PCR). The relationship between the expression of FAT1 and HIF-1α was analyzed. The expression of FAT1 and HIF-1α in human glioma cells U87MG and GOS3 under the condition of normal oxygen (20% O2) and hypoxia (1% O2) was detected by FQ-PCR. FAT1 small interfering RNA (siRNA), HIF-1α siRNA and control siRNA were transfected into U87MG and GOS3 cell lines respectively by Lipofectamine 2000. The expression of FAT1 and HIF-1 under the normal oxygen and hypoxia conditions was detected. The invasive abilities of U87MG and GOS3 glioma cells were examined by cell chamber technique. Results The relative expression levels of FAT1 and HIF-1α in 48 cases of pathological tissues of patients with grade Ⅲ-Ⅳ glioma were 4.07 (0.72, 13.42) and 0.94 (0.29, 3.49) respectively. Spearman analysis showed that there was significant correlation between the relative expression levels of FAT1 and HIF-1α (r=0.835, P=0.000). The normal oxygen concentration in GOS3 cell lines FAT1 (0.29±0.03) and HIF-1α (0.34±0.09) relative expression levels were significantly lower than that of U87MG cells (1.08±0.15, 1.00±0.12; t=8.945, 7.621; P=0.010, 0.002); hypoxia condition, U87MG and GOS3 cell lines FAT1 and HIF-1α relative expression levels were significantly higher than the normal oxygen concentration (t=17.994, 8.920, 14.047, 4.037; P=0.000, 0.001, 0.004, 0.016); U87MG cells under hypoxic conditions FAT1 (4.38±0.28) and HIF-1α (2.76±0.32) relative expression levels were significantly higher than that of GOS3 cell line (1.69±0.17, 0.83±0.19; t=14.224, 8.982; P=0.000, 0.001). After transfection of FAT1 siRNA, FAT1 (0.17±0.06, 0.16±0.03, 0.16±0.03, 0.18±0.03) and HIF-1α (0.18±0.03, 0.17±0.04, 0.15±0.04, 0.21±0.02) relative expression levels of U87MG and GOS3 cells in the normal oxygen and hypoxia were significantly decreased (t=7.577, 7.793, 7.793, 7.783, 9.285, 9.260, 9.484, 9.042; P=0.002, 0.013, 0.013, 0.014, 0.009, 0.007, 0.007, 0.011). After transfection of HIF-1α siRNA, FAT1 (0.16±0.04, 0.28±0.03) relative expression levels in the normal oxygen and HIF-1α (0.15±0.03, 0.16±0.02, 0.15±0.03, 0.17±0.02) relative expression levels of U87MG and GOS3 cells in normal oxygen and hypoxia conditions were significantly decreased (t=7.890, 6.834, 9.624, 9.614, 9.624, 9.500; P=0.012, 0.018, 0.008, 0.009, 0.008, 0.010); but the level under hypoxic conditions relative expression of FAT1 (4.27±0.41, 2.72±0.25) were significantly higher than that in normal oxygen conditions (t=12.649, 9.671; P=0.000, 0.001). Under hypoxic conditions, the invasion cell quantity of U87MG cells transfected with FAT1 siRNA and HIF-1α siRNA [(23.2±4.2) and (21.5±3.1) cells] were lower than those of normal oxygen conditions [(57.6±8.1) and (58.7±7.6) cells; t=6.530, 7.850; P=0.003, 0.001]. Conclusion The expression of FAT1 and HIF-1 in high grade gliomas is strongly correlated, and the expression is high under the hypoxia condition. FAT1 can promote the invasion of glioma cells by targeting HIF-1α. Key words: Glioma; Fat atypical cadherin; Hypoxia inducible factor-1α; Invasion

DNA methylation is related to the occurrence of breast cancer and is not affected by culture conditions.

The present study aimed to explore the relationship between DNA methylation and breast cancer under different cell culture conditions. MCF‑7 breast cancer cells were cultured in two‑dimensional (2D), three‑dimensional (3D) and orthotopic transplantation (Ti) adhesion substrates. Principal component analysis (PCA) was used for global visualization of these three samples. The methylation status of CpG sites was examined by unsupervised clustering analysis. Scatter plots and histograms were constructed from the mean β‑values from 3Dvs.2D, 3Dvs.Ti and Tivs.2D analysis. In addition, analyses of Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were conducted to explore the putative biological functions in which mutL homolog (MLH), phosphatase and tensin homolog (PTEN), runt‑related transcription factor (RUNX), Ras association domain family (RASSF), cadherin 1 (CDH1), O‑6‑methylguanine‑DNA methyltransferase (MGMT) and P16 may serve a role. Quantitative methylation‑specific polymerase chain reaction (QMSP) was performed to determine the influence of culturing conditions on important gene expression. Results from PCA analysis indicated that the three samples were closely connected with each other. Venn diagrams revealed that certain differential methylation positions were common among the three sample groups, and 116 CpG positions were identified that appeared to be hypermethylated. The methylation patterns were more similar between 3D vs. 2D cultures compared with those between 3Dvs.Ti or between Ti vs. 2D. Results of GO term and KEGG pathway analyses indicated that genes were enriched in four pathways, including transporter activity and G‑protein coupled receptor activity. In addition, QMSP analysis identified no notable differences in the methylation status of MLH, PTEN, RUNX, RASSF, CDH1, MGMT and P16 under 2D, 3D and Ti culture conditions. In conclusion, abnormal DNA methylation is related with breast cancer, and the methylation status did not change in breast cancer cells cultured in different conditions.

MicroRNA-219-5p Promotes Tumor Growth and Metastasis of Hepatocellular Carcinoma by Regulating Cadherin 1.

MicroRNAs play significant roles in the development of cancer and may serve as promising therapeutic targets. In our previous work, miR-219-5p was identified as one of the important metastasis-related microRNAs in HCC. Here we demonstrated that miR-219-5p expression was elevated in HCC tissues and was associated with vascular invasion and dismal prognosis. In multivariate analysis, miR-219-5p was identified as an independent prognostic indicator for HCC patients. Functional mechanism analyses showed that miR-219-5p promoted HCC cell proliferation and invasion in in vitro, as well as in vivo, tumor growth and metastasis in nude mice models bearing human HCC tumors. In addition, cadherin 1 (CDH1) was revealed to be a downstream target of miR-219-5p in HCC cells. In conclusion, miR-219-5p promotes tumor growth and metastasis of HCC by regulating CDH1 and can serve as a prognostic marker for HCC patients.

Molecular Network Pathway Of ERBB In Diffuse-Type Gastric Cancer, Mesenchymal Stem Cells And Epithelial-Mesenchymal Transition

Epithelial-mesenchymal transition (EMT) is related to malignancy and metastasis in cancer. The molecular networks including tyrosine kinases are altered in gastric cancer (GC). This study aims to reveal the role of ERBBs (erb-b2 receptor tyrosine kinases) in EMT, and generate the molecular network pathway of ERBBs in diffusetype GC and mesenchymal stem cells (MSCs). The expression of ERBB genes was analyzed in MSCs and diffuse-type GC which has mesenchymal characteristics compared to intestinal-type GC. The signaling and molecular network of ERBB was analyzed using several databases, including cBioPortal for Cancer Genomics, Kyoto Encyclopedia of Genes and Genomes (KEGG), BioGRID and VaProS. ERBB2 and ERBB3 gene expression were up-regulated in diffuse-type GC compared to MSCs. The ERBB3 molecular network includes epidermal growth factor receptor (EGFR), cadherin 1 (CDH1), catenin beta 1 (CTNNB1) and EPH receptor A5 (EPHA5). These results demonstrate the importance of the ERBB network in cancer signaling, and revealed a ERBB3 network pathway model in diffuse-type GC and MSCs, and EMT.

Molecular pathway network of EFNA1 in cancer and mesenchymal stem cells

Abundant molecules are dynamically activated in cancer and stem cells. To investigate the role of ephrin A1 (EFNA1) in cancer and stem cell signaling pathways, we analyzed the gene expression and molecular network of EFNA1 in mesenchymal stem cells (MSCs) and diffuse-type gastric cancer (GC). Diffuse-type GC has more mesenchymal-like feature and malignant characteristics compared to intestinal-type GC. The signaling and molecular network of EFNA1 in cancer and stem cells were analyzed using several databases, including cBioPortal for Cancer Genomics, Kyoto Encyclopedia of Genes and Genomes (KEGG). The gene expression of EFNA1 was up-regulated in diffuse-type GC compared to MSCs. The molecular pathway network of EFNA1 includes cadherin 1 (CDH1), catenin beta 1 (CTNNB1), ras-related C3 botulinum toxin substrate 1 (rho family, small GTP binding protein Rac1) (RAC1), EPH receptor A5 (EPHA5), and the KRAS proto-oncogene, GTPase (KRAS). We summarized molecular pathway network of EFNA1 in cancer and stem cells. The results revealed a network model for EFNA1 in cancer and stem cells.

Pathomorphology of the carcinomas

The paper describes the morphological and functional differentiation of cancer cells and mentions basic markers combined with that phenomenon. Table 1 presents cariotypic and immunophenotypic changes, neoplastic biomarkers and biological products present within the cells in selected cancer types. The authors also present changes in the cell cycle that leads to cancerogenesis with an emphasis put on the role of so-called genome guardians, i.e. TP53 and RB1 genes, and describe the main epigenetic factors, including the DNA methylation process in CDH1 (cadherin1) gene. The article also shows the morphologic types of proliferative changes: pre-cancer lesions (laesio praecancerosus), pre-cancer states (status praecancerosus) and pre-invasive carcinoma (carcinoma praeinvasivum, carcinoma in situ). A new classification of carcinomas is presented, including tumours originating from: a – a luminal epithelial-like cell line (with typical epithelial markers – E-cadherin, desmoplakin 1), b – a weakly luminal epithelial-like cell line (with a visibly weakened expression of epithelial antigenes) and c – a mesenchymal-like cell line (with the presence of proteins typical for mesenchymal cells – vimentin, N-cadherin, and lack of epithelial-specific antigens). Moreover, the authors extensively describe the so-called epithelial mesenchymal transition (EMT) that can be observed both in in vitro and in vivo conditions. The role of cancer-associated fibroblasts (CAFs) in that process is shown. The cells exhibit an increase in the expression of genes involved in adhesion and angiogenesis and an increased expression of neurotransmitter receptors (adrenaline, noradrenaline)....

Transcriptome profiling of cancer tissues in Chinese patients with gastric cancer by high-throughput sequencing.

Gastric cancer (GC) is the fifth most common malignancy and the third leading cause of cancer-associated mortality worldwide. Therefore, there is a requirement to identify sufficiently sensitive biomarkers for GC. Genome-wide screening of transcriptome dysregulation among cancerous and normal tissues may provide insights into the underlying molecular mechanisms of GC initiation and progression. At present, high-throughput sequencing techniques have begun to innovate biomedical studies. The RNA-seq method has become an advanced approach in medical studies; it is capable of the accurate detection of gene expression levels. The present study used RNA-seq to evaluate the transcriptional changes between tumor and matched normal samples, and these changes were confirmed by differentially expressed genes in larger samples using the results of sequencing. In total, the upregulation of 28 mRNAs and downregulation of 22 mRNAs between cancerous and normal tissue samples were identified. Subsequently, five differentially expressed genes were selected to verify in large samples and cadherin-1 (CDH1) was selected to detect protein expression levels. The results revealed that CDH1, cyclooxygenase-2 and matrix metalloproteinase genes had significantly higher expression levels, whereas the expression levels of dermatopontin and transforming growth factor β receptor 2 were decreased in GC samples. In particular, CDH1 demonstrated a 36-fold higher expression level in cancer tissues. The western blotting results also revealed high CDH1 expression levels in the validation cohorts. Furthermore, these genes are highly enriched in certain gene ontology categories, including the digestive system process, secretion and digestion. The present study provided a preliminary survey of the transcriptome of Chinese patients with GC, which may improve the detection of aberrant gene expression in GC and the understanding of the mechanisms of tumorigenesis.

Label-free Detection for a DNA Methylation Assay Using Raman Spectroscopy.

Background:DNA methylation has been suggested as a biomarker for early cancer detection and treatment. Varieties of technologies for detecting DNA methylation have been developed, but they are not sufficiently sensitive for use in diagnostic devices. The aim of this study was to determine the suitability of Raman spectroscopy for label-free detection of methylated DNA.Methods:The methylated promoter regions of cancer-related genes cadherin 1 (CDH1) and retinoic acid receptor beta (RARB) served as target DNA sequences. Based on bisulfite conversion, oligonucleotides of methylated or nonmethylated probes and targets were synthesized for the DNA methylation assay. Principal component analysis with linear discriminant analysis (PCA-DA) was used to discriminate the hybridization between probes and targets (methylated probe and methylated target or nonmethylated probe and nonmethylated target) of CDH1 and RARB from nonhybridization between the probe and targets (methylated probe and nonmethylated target or nonmethylated probe and methylated target).Results:This study revealed that the CDH1 and RARB oligo sets and their hybridization data could be classified using PCA-DA. The classification results for CDH1 methylated probe + CDH1 methylated target versus CDH1 methylated probe + CDH1 unmethylated target showed sensitivity, specificity, and error rates of 92%, 100%, and 8%, respectively. The classification results for the RARB methylated probe + RARB methylated target versus RARB methylated probe + RARB unmethylated target showed sensitivity, specificity, and error rates of 92%, 93%, and 11%, respectively.Conclusions:Label-free detection of DNA methylation could be achieved using Raman spectroscopy with discriminant analysis.

Similarity in gene-regulatory networks suggests that cancer cells share characteristics of embryonic neural cells

Cancer cells are immature cells resulting from cellular reprogramming by gene misregulation, and redifferentiation is expected to reduce malignancy. It is unclear, however, whether cancer cells can undergo terminal differentiation. Here, we show that inhibition of the epigenetic modification enzyme enhancer of zeste homolog 2 (EZH2), histone deacetylases 1 and 3 (HDAC1 and -3), lysine demethylase 1A (LSD1), or DNA methyltransferase 1 (DNMT1), which all promote cancer development and progression, leads to postmitotic neuron-like differentiation with loss of malignant features in distinct solid cancer cell lines. The regulatory effect of these enzymes in neuronal differentiation resided in their intrinsic activity in embryonic neural precursor/progenitor cells. We further found that a major part of pan-cancer-promoting genes and the signal transducers of the pan-cancer-promoting signaling pathways, including the epithelial-to-mesenchymal transition (EMT) mesenchymal marker genes, display neural specific expression during embryonic neurulation. In contrast, many tumor suppressor genes, including the EMT epithelial marker gene that encodes cadherin 1 (CDH1), exhibited non-neural or no expression. This correlation indicated that cancer cells and embryonic neural cells share a regulatory network, mediating both tumorigenesis and neural development. This observed similarity in regulatory mechanisms suggests that cancer cells might share characteristics of embryonic neural cells.

MicroRNA-1285-5p influences the proliferation and metastasis of non-small-cell lung carcinoma cells via downregulating CDH1 and Smad4

Abnormal expression of microRNAs has been reported to regulate gene expression and cancer cell growth, invasion, and migration. Recently, upregulation of hsa-miR-1285 was demonstrated in bronchoalveolar lavage fluid samples from patients with lung cancer and downregulation in plasma level of stage-I lung cancer patients. However, the function and the underlying mechanism of miR-1285 in non-small-cell lung carcinoma have not been elucidated. In this study, we found that miR-1285-5p, the mature form of miR-1285, was significantly upregulated in human non-small-cell lung carcinoma cell lines A549 and SK-MES-1. Additionally, cells transfected with the miR-1285-5p inhibitor LV-anti-miR-1285-5p demonstrated significantly inhibited proliferation and invasion and depressed migration. Further analysis demonstrated that the miR-1285-5p precursor LV-miR-1285-5p attenuated the expression of Smad4 and cadherin-1 (CDH1) but that LV-anti-miR-1285-5p showed opposite results. A luciferase reporter assay confirmed that miR-1285-5p targeted Smad4 and CDH1. Mechanism analyses revealed that silence of Smad4 and CDH1 significantly attenuated the inhibitory effects of LV-anti-miR-1285-5p on non-small-cell lung carcinoma growth and invasion. Taken together, our data suggest that miR-1285-5p functions as a tumor promoter in the development of non-small-cell lung carcinoma by targeting Smad4 and CDH1, indicating a novel therapeutic strategy for non-small-cell lung carcinoma patients.

Maternal separation induces hippocampal changes in cadherin-1 (CDH-1) mRNA and recognition memory impairment in adolescent mice

In rodents, disruption of mother-infant attachment induced by maternal separation (MS) is associated with recognition memory impairment and long-term neurobiological consequences. Particularly stress-induced modifications have been associated to disruption of cadherin (CDH) adhesion function, which plays an important role in remodeling of neuronal connection and synaptic plasticity. This study investigated the sex-dependent effect of MS on recognition memory and mRNA levels of classical type I and type II CDH and the related β -catenin (β -Cat) in the hippocampus and prefrontal cortex of late adolescent mice. We provided evidence that the BALB/c mice exposed to MS present deficit in recognition memory, especially females. Postnatal MS induced higher hippocampal CDH-2 and CDH-8 mRNA levels, as well as an upregulation of CDH-1 in the prefrontal cortex in both males and females. MS-reared female mice presented lower CDH-1 mRNA levels in the hippocampus. In addition, hippocampal CDH-1 mRNA levels were positively correlated with recognition memory performance in females. MS-reared male mice exhibited higher β -Cat mRNA levels in the hippocampus. Considering sex-specific effects on CDH mRNA levels, it has been demonstrated mRNA changes in CDH-1, β -Cat, and CDH-6 in the hippocampus, as well as CDH-1, CDH-8 and CDH-11 in the prefrontal cortex. Overall, these findings suggest a complex interplay among MS, CDH mRNA expression, and sex differences in the PFC and hippocampus of adolescent mice.

Cadherin 1 protein expression level in esophageal cancer and its relationship with clinical pathological characteristics and prognosis

Objective To explore the cadherin 1 (Cdh1) protein expression level in esophageal cancer and its relationship with clinical pathological characteristics and prognosis. Methods Retrospectively analysis the clinical data of 132 patients of esophageal cancer surgery in our hospital. 132 cases esophageal cancer tissue (observation group) and tissue adjacent to carcinoma (control group) were selected as the research object. Cdh1 mRNA and protein expression level in control group and observation group were analyzed by real-time fluorescent quantitative PCR, Western blotting and immunohistochemistry. The relationship of Cdh1 between clinical features and prognosis were analyzed. Results Compared with control group, Cdh1 mRNA and protein levels in the observation group significantly reduced, the difference was statistically significant (P=0.004, P=0.009). Immunohistochemical analysis showed that Cdh1 protein is mainly expressed in cell membrane of esophageal tissue. Cdh1 protein expression positive rate in the control group and observation group were 100% and 21.96%, respectively, the difference was statistically significant (P=0.021). In esophageal cancer patients, the expression rate of Cdh1 was correlated with the degree of differentiation of esophageal cancer, TMN stage, invasion depth and lymph node metastasis of esophageal cancer and was not correlated withpatient’s age, gender, nationality. In the observation group, the 1, 3 years of cumulative survival rate in Cdh1 positive group significantly increased than that of negative group (P=0.012). Conclusion Cdh1 protein expression level decreased obviously in esophageal cancer, and the degree of the patients with lymph node metastasis, invasion, TNM stage and prognosis is closely related, can be used as auxiliary diagnostic marker of prognosis. Key words: Esophageal cancer; Cadherin 1 protein; Clinical pathological features; Prognosis

Promoter methylation status of the tumor suppressor genes p16 and cadherin 1 in cervical intraepithelial neoplasia.

Cervical cancer is the second most common female cancer worldwide. DNA methylation is one of a number of epigenetic regulation mechanisms leading to gene silencing in neoplastic cells. Aberrant methylation results in the silencing of tumor suppressor gene expression, and has been detected in a high percentage of human cancers. In the present study, the methylation status of three tumor suppressor genes, retinoic acid receptor β (RARβ), p16 and cadherin 1 (CDH1), and the inflammatory-associated cyclooxygenase-2 (COX-2) gene, was examined at distinct stages of cervical intraepithelial neoplasia (CIN). The results of the present study revealed that the COX-2 gene was unmethylated between CIN I and carcinoma specimens. The RARβ gene exhibited a minimal change in methylation frequency, whereas the CDH1 methylation level was increased <2-fold between CIN I and carcinoma. Notably, the methylation frequency of p16 was 13.2% in normal specimens; 18.2% in CIN I; 35.7% in CIN II; 31.6% in CIN III; and 15.4% in carcinoma. By contrast, the methylation frequency of p16 increased between CIN I and carcinoma in the absence of high-risk group papillomaviruses. The results of bisulfite sequencing indicated that the 10 CpG sites were all methylated in p16 gene methylation-positive individuals. The results of the present study demonstrate that the methylation frequency of p16 and CDH1 was progressively increased during the development of malignant stages in CIN, and may be an additional tool for current cytomorphology-based screening of cervical cell specimens.

Circulating-free DNA Mutation Associated with Response of Targeted Therapy in Human Epidermal Growth Factor Receptor 2-positive Metastatic Breast Cancer.

Background:The addition of anti-human epidermal growth factor receptor 2 (HER2)-targeted drugs, such as trastuzumab, lapatinib, and trastuzumab emtansine (T-DM1), to chemotherapy significantly improved prognosis of HER2-positive breast cancer patients. However, it was confused that metastatic patients vary in the response of targeted drug. Therefore, methods of accurately predicting drug response were really needed. To overcome the spatial and temporal limitations of biopsies, we aimed to develop a more sensitive and less invasive method of detecting mutations associated with anti-HER2 therapeutic response through circulating-free DNA (cfDNA).Methods:From March 6, 2014 to December 10, 2014, 24 plasma samples from 20 patients with HER2-positive metastatic breast cancer who received systemic therapy were eligible. We used a panel for detection of hot-spot mutations from 50 oncogenes and tumor suppressor genes, and then used targeted next-generation sequencing (NGS) to identify somatic mutation of these samples in those 50 genes. Samples taken before their first trastuzumab administration and subsequently proven with clinical benefit were grouped into sensitive group. The others were collected after disease progression of the trastuzumab-based therapy and were grouped into the resistant group.Results:A total of 486 single-nucleotide variants from 46 genes were detected. Of these 46 genes, phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), proto-oncogene c-Kit (KIT), and tumor protein p53 (TP53) were the most common mutated genes. Seven genes, including epidermal growth factor receptor (EGFR), G protein subunit alpha S (GNAS), HRas proto-oncogene (HRAS), mutL homolog 1 (MLH1), cadherin 1 (CDH1), neuroblastoma RAS viral oncogene homolog (NRAS), and NOTCH1, that only occurred mutations in the resistant group were associated with the resistance of targeted therapy. In addition, we detected a HER2 S855I mutation in two patients who had persistent benefits from anti-HER2 therapy.Conclusion:Targeted NGS of cfDNA has potential clinical utility to detect biomarkers from HER2-targeted therapies.