Expression Pattern Of N-cadherin Research Articles

Iodine-125 seed brachytherapy inhibits non–small cell lung cancer by suppressing epithelial-mesenchymal transition

PurposeMetastatic non–small cell lung cancer (NSCLC) has posed a great clinical challenge with high mortality. Iodine-125 (I-125) seed brachytherapy has not been widely applied in clinic against NSCLC. Methods and MaterialsMice were injected with human H23 NSCLC cells to establish a mouse xenograft model, which received I-125 seed implantation. The curative effect, pathological impairments, and survival rate of mice were investigated. Changes in the expression levels of epithelial-mesenchymal transition (EMT) markers, including N-cadherin, E-cadherin, and vimentin, in the xenograft tumors were analyzed using reverse transcription polymerase chain reaction and Western blot. ResultsThe tumor volume and pathological effect were reduced by I-125 seed implant. I-125 seed implant also significantly improved survival rate of the model mice. Expression patterns of N-cadherin, E-cadherin, and vimentin were reversed in I-125 seed–implanted mice in comparison with control mice, indicating suppressed EMT. ConclusionsI-125 seed brachytherapy significantly inhibits NSCLC by suppressing EMT in a mouse model.

Abstract 2620: Progressive expression of n-cadherin and epidermal growth factor receptor in colorectal adenocarcinoma.

Abstract Purpose: N-cadherin (NCAD) and epidermal growth factor receptor (EGFR) have been found to be inducers of epithelial-to-mesenchymal transition that are known to play a role in invasion and metastasis of cancer. Over expression of NCAD and EGFR are associated with increased tumor invasiveness. However, their progressive expression in colorectal cancer (CRC) has not been studied. Therefore, in the present study, we investigated expression status of NCAD and EGFR in paired lesions of CRC. Experimental Design: We examined the expression of NCAD and EGFR in paired lesions of human CRC (hCRC) and human mice CRC (hmCRC) xenograft by immunofluorescence (IF). Images were acquired using the Olympus FluoView FV1000 (OFVFV1000) Confocal Microscope. The average FITC fluorescence intensity (aFFI) was measured by OFVFV1000 software. Western blot analysis was performed to validate results obtained from IF method. Results: Differential expression pattern of NCAD was observed in paired normal, primary and metastatic lesions of hCRC and hmCRC xenograft tissues. For NCAD, it was mainly localized in cytoplasm (diffused), nucleus (moderate levels), and membrane or cell junctions of CRC. For EGFR, normal colon tissue or primary CRC showing membranous EGFR-low positive staining. In contrast corresponding liver metastasis showing membranous EGFR-high positive staining. The NCAD and EGFR expressions were correlated with metastasis phenotype of hCRC. The expressions of NCAD ((aFFI; 812 vs. 261) and EGFR (aFFI 421 vs.131) were each found to be 3 times higher in liver metastasis of hCRC as compared to primary tumor. It has been observed that the expression of NCAD was substantially increased in metastatic lesion of hmCRC xenograft as compared to primary lesion of hmCRC xenograft. Conclusion: This study revealed that upregulation of NCAD and EGFR expressions is a mechanism by which CRC lesions acquire a more aggressive phenotype and, thereby, progress to a more advanced stage of CRC. Furthermore, our findings are likely to have implications in the design of clinical trials targeting the NCAD and EGFR proteins in CRC. Citation Format: Venkat R. Katkoori, Upender Manne, Marc D. Basson, Harvey L. Bumpers. Progressive expression of n-cadherin and epidermal growth factor receptor in colorectal adenocarcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2620. doi:10.1158/1538-7445.AM2013-2620

Abstract 2320: N-cadherin down-regulation has antitumor effects in esophageal cancer

Abstract Tumor microenviroment plays a critical role for cancer and their malignancy. Dynamic changes in tumor microenvironment have been observed in many tumors including esophageal cancer. And during the drastic change, some of tumor cells show EMT (epithelial mesenchymal transition) which means that epithelial cells lose their epithelial feature and acquire mesenchymal properties, as invasiveness and also communication with other kinds of mesenchymal cells. During those change, one of the marker of epithelial feature, E-cadherin is switch to N-cadherin, it has been known as “cadherin switch”. And it is thought that those changes have relations with cancer malignancy. Previously we reported that esophageal cancer cells secrete TGF-b1 and activate mesenchymal fibroblasts, and those activated fibroblasts guided more endothelial cells into tumor and induced angiogenesis. In the other hand, we hypothesis cell-cell direct communication like cell-cell contact is also important role for cancer progression. So in this report, firstly we analyze the expression of cadherins in surgical specimens in immunohistochemistry and determined expression patterns of N-cadherin in esophageal squamous cell carcinoma and Barrett's adenocarinoma. Some of those cancers showed N-cadherin expression and specifically expressed in their invasive front. Secondary we analyzed cancer cell lines as well, TE-1, TE-4, TE-10, OE19 and OE33. Those cell lines are squamous cell carcinoma cell lines and Barrett's adenocarcinoma cell lines, and those showed expression of E-cadherin and some of them showed expression of N-cadherin. And we used those cell lines and explored the behavioral differentiations of each cell lines. Here, we report and show analysis of E- or N- cadherin expression in surgical specimens and cancer cell lines. And also we could show the potential of regulation of cadherins in esophageal cancer as a new therapeutic target, which is well recognized as one of the most aggressive malignancies in the world. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2320.

Effect of N-cadherin knock-down on invasiveness of esophageal squamous cell carcinoma

e15571 Background: Cell adhesion molecules are of crucial importance in cancer invasion and metastasis. Epithelial to mesenchymal transition, characterized by reduced E-cadherin and increased N-cadherin expression, has been recognized as a feature of aggressive tumors, but the importance of this phenotype has not been settled in esophageal squamous cell carcinoma. Aim: To examine the expressions of N-cadherin and E-cadherin in 62 normal esophageal epithelium specimens, 31 adjacent atypical hyperplasia epithelium specimens and 62 esophageal squamous cell carcinoma specimens, and to investigate the roles of N-cadherin in the invasiveness of esophageal squamous cell carcinoma cell line EC9706 transfected by N-cadherin shRNA.. Methods: PV immunohistochemistry was used to detect the expression pattern of N-cadherin and E-cadherin in 62 normal esophageal epithelium specimens, 31 adjacent atypical hyperplasia epithelium specimens and 62 esophageal squamous cell carcinoma specimens. The invasiveness of esophageal squamous cell carcinoma cell line EC9706 in vitro and in vivo was determined by transwell assay and nude mice experiments after EC9706 was transfected by N-cadherin shRNA. Results: The positive rates of N-cadherin decreased in the sequence of carcinoma, adjacent atypical hyperplasia and normal esophageal tissue, which were 75.8%, 61.3%, 29.0% (P < 0.05), respectively, while those of E-cadherin increased in sequence, which were 40.3%, 71.0% and 95.2% (P < 0.05). The increased expression of N-cadherin and decreased expression of E-cadherin were related to the invasion, differentiation, and lymph node metastasis (P < 0.05). The expression level of N-cadherin decreased in the N- cadherin knocked down cells, and the invasiveness of those cells decreased significantly as well in vitro and in vivo. Conclusions: These results suggest that N-cadherin is an important factor in the invasiveness of esophageal squamous cell carcinoma and N-cadherin may serves as a potential molecular target for biotherapy of esophageal squamous cell carcinoma. No significant financial relationships to disclose.

N‐cadherin expression in palisade nerve endings of rat vellus hairs

Palisade nerve endings (PNs) are mechanoreceptors around vellus hairs of mammals. Each lanceolate nerve ending (LN) of the PN is characterized by a sensory nerve ending symmetrically sandwiched by two processes of type II terminal Schwann cells (tSCIIs). However, the molecular mechanisms underlying the structural organization of the PN are poorly understood. Electron microscopy showed that adherens junctions appeared to adhere to the sensory nerve ending and tSCII processes, so we examined the location of the N-cadherin adhesion system in PNs of rat vellus hairs by using immunoelectron microscopy. N-cadherin localized near both ends of the cell boundary between sensory nerve ending and tSCII processes, which corresponded to the sites of adherens junctions. We further found cadherin-associated proteins, alpha- and beta-catenins, at the linings of adherens junctions. Three-dimensional reconstruction of immunoelectron microscopic serial thin sections showed four linear arrays of N-cadherin arranged longitudinally along the LN beneath the four longitudinal borders of two tSCII processes. In contrast, sensory nerve fibers just proximal to the LNs formed common unmyelinated nerve fibers, in which N-cadherin was located mainly at the mesaxon of type I terminal Schwann cells (tSCIs). These results suggest that the four linear arrays of N-cadherin-mediated junctions adhere the sensory nerve ending and tSCII processes side by side to form the characteristic structure of the LN, and the structural differences between the LNs and the proximal unmyelinated nerve fibers possibly are due to the difference in the pattern of N-cadherin expression between sensory nerve endings and tSCII or tSCI processes.

Loss of N-cadherin and α-catenin in the proximal tubules of aging male Fischer 344 rats

Aging is associated with a loss of renal reserve, and increased sensitivity to either xenobiotic or physiologic insult. Given the critical role of the cadherin/catenin complex in establishing and maintaining the integrity and polarity of tubular epithelial cells, it was hypothesized that aging was associated with alterations in renal cadherin/catenin complexes. Histological assessment of aged (24 months) kidneys harvested from male Fischer 344 rats demonstrates mild degeneration of proximal tubules, multifocal chronic lymphocytic infiltration, moderate development of protein casts inside tubules, and tubular dilatation or degeneration. Western blot analysis revealed that N-cadherin protein expression is not constant over 24 months. N-cadherin expression increased from 4 to 9 months, with peak levels at 9 and 13 months. A decrease in expression was seen at 19 months and an almost complete loss of expression was seen at 24 months. In contrast, the expression of E- and Ksp-cadherin was constant over 24 months. A loss of α-catenin at was seen at 19 and 24 months in the absence of changes in β-, γ-, and p120-catenin. This pattern of N-cadherin expression (increase followed by decrease) was confirmed by real-time PCR analysis, which demonstrated a similar pattern as the Western blot, suggesting that the loss of N-cadherin protein was due to decreased gene expression. The loss of N-cadherin was specific for the kidney, as no changes in N-cadherin expression in the liver, brain, or testes were seen during aging. The conclusion that loss of N-cadherin expression is a critical component of the renal dysfunction associated with aging is supported by the finding that caloric restriction attenuates the loss of N-cadherin, as well as the finding that a significant loss of N-cadherin is seen in the kidneys of ZDF×SHHF rats, a genetic model of end-stage renal disease. Cadherin and catenin expression was further analyzed by immunofluorescence. A significant loss of staining of both N-cadherin and α-catenin was seen in the proximal tubules of rats at 24 months. Interestingly, this corresponded with delocalization of the α-1 subunit of the Na +K +-ATPase, i.e. aberrant staining on cell–cell borders and some indication of apical staining in proximal tubules. Taken together, these data suggest that aging is associated with decreased expression of N-cadherin and α-catenin and is associated with a loss of cell polarity.

Downregulation of N-cadherin in the neointima stimulates migration of smooth muscle cells by RhoA deactivation

The aim of the study was to analyze whether cadherin- and Rho-family GTPases-mediated dynamic rearrangement of cell-cell adhesion play an important role during human arterial smooth muscle cell (haSMC) migration. Expression patterns of N-cadherin and beta-catenin were analyzed in a domestic pig restenosis model after 14, 28, and 90 days as well as in quiescent and migratory haSMCs in vitro. N-cadherin expression was upregulated by transient sense; downregulation was induced by antisense transfection. For functional inhibition, antibody GC-4 was used. Cell migration was quantified using Boyden chamber assays. Regulation of RhoA GTPase was tested by assessment of RhoA activity. In vivo analysis of N-cadherin expression in a porcine restenosis model revealed downregulation in the neointima after 14 days. After 28 days, N-cadherin expression was slightly restored, while after 90 days, no difference between medial and neointimal expression was detectable. beta-Catenin levels remained unchanged during the whole period. According to the in vivo situation, N-cadherin was significantly downregulated in migratory haSMCs compared to quiescent cells in vitro. After N-cadherin overexpression, haSMC migration was reduced by 87% (P<0.001). By contrast, inhibition of N-cadherin in quiescent haSMCs by GC-4 increased the migratory potential by 87% (P<0.01). In haSMCs overexpressing N-cadherin, a significant upregulation of RhoA activity was demonstrated, while RhoA activity was blocked by GC-4. These results indicate that the regulation of haSMC attachment by N-cadherins is essential for haSMC migration. Modification of N-cadherin expression and activity induces RhoA signaling with relevance for the reorganization of the actin cytoskeleton.

Reduced Expression of Plakoglobin Correlates with Adverse Outcome in Patients with Neuroblastoma

Plakoglobin and its homologue beta-catenin are cytoplasmic proteins that mediate adhesive functions by interacting with cadherin receptors and signaling activities by interacting with transcription factors. It has been suggested that plakoglobin can suppress tumorigenicity whereas beta-catenin can act as an oncogene. We investigated the correlation between the expression pattern of N-cadherin, beta-catenin, and plakoglobin and tumor behavior in primary tumors of 20 neuroblastoma patients of all stages and in 11 human neuroblastoma cell lines. N-cadherin and beta-catenin were detected in 9 of 11 and 11 of 11 cell lines, respectively, whereas plakoglobin was undetectable or severely reduced in 6 of 11 cell lines. Tumor cells from 16 of 20 patients expressed N-cadherin and 20 of 20 patients expressed beta-catenin at levels similar to those of normal ganglion cells. Plakoglobin was undetectable in 9 of 20 tumors. Plakoglobin deficiency in the primary tumors was significantly associated with adverse clinical outcome. Five of the patients with plakoglobin-negative tumors died whereas four patients are alive without evident disease. In contrast, all patients with plakoglobin-positive tumors are alive; 2 of 11 are alive with the disease and 9 of 11 are alive without evident disease. These results suggest that down-regulation of plakoglobin may be of prognostic value for neuroblastoma patients as predictor of poor outcome.