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Changcheng Guo Kai Zhu Wei Sun Bin Yang Wenyu Gu Jun Luo Bo Peng Junhua Zheng 《Biochemical and biophysical research communications》2014
Objective
This study aimed at detecting Pokemon expression in bladder cancer cell and investigating the relationship between Pokemon and epithelial–mesenchymal transition. Furthermore, we investigated the functions of Pokemon in the carcinogenesis and development of bladder cancer. This study was also designed to observe the inhibitory effects of siRNA expression vector on Pokemon in bladder cancer cell.Methods
The siRNA expression vectors which were constructed to express a short hairpin RNA against Pokemon were transfected to the bladder cancer cells T24 with a liposome. Levels of Pokemon, E-cadherin and β-catenin mRNA and protein were examined by real-time quantitative-fluorescent PCR and Western blot analysis, respectively. The effects of Pokemon silencing on epithelial–mesenchymal transition of T24 cells were evaluated with wound-healing assay.Results
Pokemon was strongly inhibited by siRNA treatment, especially siRNA3 treatment group, as it was reflected by Western blot and real-time PCR. The gene and protein of E-cadherin expression level showed increased markedly after Pokemon was inhibited by RNA interference. While there were no differences in the levels of gene and protein of β-catenin among five groups. The bladder cancer cell after Pokemon siRNA interference showed a significantly reduced wound-closing efficiency at 6, 12 and 24 h.Conclusions
Our findings suggest Pokemon may inhibit the expression of E-cadherin. The low expression of E-cadherin lead to increasing the phenotype and apical-base polarity of epithelial cells. These changes of cells may result in the recurrence and progression of bladder cancer at last. 相似文献3.
The epithelial–mesenchymal transition (EMT) occurs commonly during carcinoma invasion and metastasis, but not during early
tumorigenesis. Microarray data demonstrated elevation of vimentin, a mesenchymal marker, in intestinal adenomas from Apc
Min/+ (Min) mice. We have tested the involvement of EMT in early tumorigenesis in mammalian intestines by following EMT-associated
markers. Elevated vimentin RNA expression and protein production were detected within neoplastic cells in murine intestinal
adenomas. Similarly, vimentin protein was detected in both adenomas and invasive adenocarcinomas of the human colon, but not
in the normal colonic epithelium or in hyperplastic polyps. Expression of E-cadherin varied inversely with vimentin. In addition,
the expression of fibronectin was elevated while that of E-cadherin decreased. Canonical E-cadherin suppressors, such as Snail,
were not elevated in the same tumor. Elevated vimentin expression in the adenoma was not correlated with persistent Ras signaling,
but was strongly correlated with reduced proliferation indices, active Wnt signaling, and TGF-β signaling, as demonstrated
by its dependence on Smad3. We designate our observations of expression of only some of the canonical features of EMT as “truncated EMT”. These unexpected
observations are interpreted as reflecting the involvement of a core of the EMT system during the tissue remodeling of early
tumorigenesis. 相似文献
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Julia König Ingrid Lang Monika Siwetz Julia Fröhlich Berthold Huppertz 《Cell and tissue banking》2014,15(2):193-198
The amnionic membrane is a rich source of multipotent mesenchymal stromal cells (hAMSC), which are readily available and show a potential use in regenerative medicine and tissue engineering. Before these cells can be applied clinically, careful characterization is necessary, especially as primary cells are known to change their phenotype in culture. We analyzed the mesenchymal phenotype of hAMSC at different stages after isolation using immunohistochemistry. Shortly after isolation (1 day), 92 % (±7 %) of the hAMSC expressed the mesenchymal marker vimentin, 2 % (±1 %) stained for the epithelial marker cytokeratin-7 and 5 % (±4 %) co-expressed these markers. After 5 days, the double positive cells slightly increased to 7 % (±3 %), while exclusive expression of cytokeratin-7 or vimentin remained unchanged (1 % ± 2 % and 92 % ± 1 %, respectively). After the first passage, all attached cells were vimentin-positive, while 54 % (±9 %) co-expressed cytokeratin-7 and vimentin. Thus, we conclude that under culture, hAMSC adopt a hybrid mesenchymal–epithelial phenotype. It is also essential to perform microscopical examination during the first days after isolation to detect contaminations with human amnion-derived epithelial cells in cultures of hAMSC. 相似文献
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Xuan Feng Zhi Wei Sai Zhang Jiayi Zhou Jing Wu Baoxin Luan Yan Du Hongbo Zhao 《Acta biochimica et biophysica Sinica》2021,(2)
Laeverin(LVRN)was first detected on the outer layer of the chorion laeve and migrating extravillous trophoblasts(EVTs).It is an enzyme that plays an important role in the placentation and pathophysiology of preeclampsia(PE).Previous studies have indicated that LVRN may be required for the invasion of human trophoblast cells.Paradoxically,LVRN was found to be highly expressed in the trophoblasts of PE patients with impaired invasive capacities.In this study,we detected the expression of LVRN in the placentas of PE patients(n=5)and normal term pregnancy women(n=5)as a control group by immunohistochemistry.LVRN was elevated in decidua(P=0.0083)and villi(P=0.0079)of PE patients.Next,LVRN was overexpressed via adeno-associated virus-mediated gene transfer in trophoblastic cell lines HTR8,Swan71,and JAR.Matrigel transwell assay and wound healing assay showed that overexpression of LVRN impeded the invasion of these three cell lines.Western blot analysis showed that LVRN overexpression caused downregulation of N-cadherin and vimentin and upregulation of E-cadherin,suggesting the inhibitory role of LVRN in epithelial–mesenchymal transition(EMT).Moreover,our data indicated that long noncoding RNA NONSTAT103348(lnc10-7)was elevated in PE patients.Silencing lnc10-7 led to decreased LVRN expression.Taken together,although the basal level of LVRN may be crucial for cell invasion,overexpression of LVRN may abrogate the cell invasiveness,suggesting a multifaceted role of LVRN in the pathogenesis of PE. 相似文献
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Epithelial–mesenchymal transition (EMT) is a crucial step in tumor progression and has an important role during cancer invasion and metastasis. Although fucosyltransferase IV (FUT4) has been implicated in the modulation of cell migration, invasion and cancer metastasis, its role during EMT is unclear. This study explores the molecular mechanisms of the involvement of FUT4 in EMT in breast cancer cells. Breast cancer cell lines display increased expression of FUT4, which is accompanied by enhanced appearance of the mesenchymal phenotype and which can be reversed by knockdown of endogenous FUT4. Moreover, FUT4 induced activation of phosphatidylinositol 3-kinase (PI3K)/Akt, and inactivation of GSK3β and nuclear translocation of NF-κB, resulting in increased Snail and MMP-9 expression and greater cell motility. Taken together, these findings indicate that FUT4 has a role in EMT through activation of the PI3K/Akt and NF-κB signaling systems, which induce the key mediators Snail and MMP-9 and facilitate the acquisition of a mesenchymal phenotype. Our findings support the possibility that FUT4 is a novel regulator of EMT in breast cancer cells and a promising target for cancer therapy. 相似文献
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Rajib Dhar Arikketh Devi Sukhamoy Gorai Saurabh Kumar Jha Athanasios Alexiou Marios Papadakis 《Journal of cellular and molecular medicine》2023,27(11):1603-1607
This short communication will enlighten the readers about the exosome and the epithelial-mesenchymal transition (EMT) related to several complicated events. It also highlighted the therapeutic potential of exosomes against EMT. Exosome toxicology, exosome heterogeneity, and a single exosome profiling approach are also covered in this article. In the future, exosomes could help us get closer to cancer vaccine and precision oncology. 相似文献
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Fan Li Xuedong Yin Xinrong Luo Hong-Yuan Li Xinliang Su Xiao-Yi Wang Li Chen Ke Zheng Guo-Sheng Ren 《Cellular signalling》2013,25(6):1413-1422
The inhibitor of apoptosis proteins (IAP) are closely correlated with proliferation, apoptosis, motility, and metastasis. Livin is the most recently identified IAP, and its role in breast progression remains unknown. In our study, analyses of 50 patients with breast cancer revealed that the positive expression rate of Livin was higher in breast cancer tissues (62%) relative to that in adjacent (35%) and normal tissues (25%). Livin expression in breast cancer correlated with the clinical stage and axillary lymph node metastasis and could be used as a prognostic marker. Our in vitro experiment revealed that Livin was highly expressed in high-invasive MDA-MB-231 cells as compared to low-invasive cells (MCF-7). Suppression of Livin by short-hairpin RNA reduced the Livin expression of MDA-MB-231 cells and subsequently inhibited tumor cell growth, proliferation, and colony formation and induced tumor cell apoptosis, motility, migration, and invasion. Overexpression of Livin in MCF7 cells resulted in increased migration and invasion capabilities of the cells without affecting proliferation and apoptosis. In addition, epithelial–mesenchymal transition (EMT) was induced by Livin expression in breast cancer cell lines. The high level of phosphorylated AKT in MDA-MB-231 cells was suppressed by Livin knockdown. Further, Livin-induced migration and invasion could be abolished by either the application of the phosphoinositide-3-kinase inhibitor LY294002 or knockdown of AKT expression using small-interfering RNA. In conclusion, Livin serves as an independent prognostic indicator for breast cancer. Livin expression promotes breast cancer metastasis through the activation of AKT signaling and induction of EMT in breast cancer cells both in vitro and in vivo. 相似文献
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You Hong Xu Jian Qin Xiaochun Qian Guodong Wang Yang Chen Fulei Shen Xiaoxu Zhao Dong Liu Qi 《Journal of molecular histology》2021,52(4):823-838
Journal of Molecular Histology - The aim of the present study was to investigate the role and potential regulatory mechanisms of fascin in the invasion and epithelial-to-mesenchymal transition of... 相似文献
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Evaluation of morphological changes in cells is an integral part of study on epithelial to mesenchymal transition (EMT), however, only a few papers reported the changes in quantitative parameters and no article compared different parameters for demanding better parameters. In the study, the purpose was to investigate suitable parameters for quantitative evaluation of EMT morphological changes. A549 human lung adenocarcinoma cell line was selected for the study. Some cells were stimulated by transforming growth factor-β1 (TGF-β1) for EMT, and other cells were as control without TGF-β1 stimulation. Subsequently, cells were placed in phase contrast microscope and three arbitrary fields were captured and saved with a personal computer. Using the tools of Photoshop software, some cells in an image were selected, segmented out and exchanged into unique hue, and other part in the image was shifted into another unique hue. The cells were calculated with 29 morphological parameters by Image Pro Plus software. A parameter between cells with or without TGF-β1 stimulation was compared statistically and nine parameters were significantly different between them. Receiver operating characteristic curve (ROC curve) of a parameter was described with SPSS software and F-test was used to compare two areas under the curves (AUCs) in Excel. Among them, roundness and radius ratio were the most AUCs and were significant higher than the other parameters. The results provided a new method with quantitative assessment of cell morphology during EMT, and found out two parameters, roundness and radius ratio, as suitable for quantification. 相似文献
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Feng Du Si Li Tian Wang Hai-Yan Zhang De-Tian Li Zhen-Xian Du Hua-Qin Wang 《Experimental biology and medicine (Maywood, N.J.)》2015,240(5):566-575
The epithelial–mesenchymal transition (EMT) of tubular epithelial cells to myofibroblast-like cells plays a substantial role in renal tubulointerstitial fibrosis, which is a common pathological character of end-stage renal disease (ESRD). Fibroblast growth factor-2 (FGF-2) triggers EMT in tubular epithelial cells and increases Bcl-2-associated athanogene 3 (BAG3) expression in neural progenitor and neuroblastoma cells. In addition, a novel role of regulation of EMT has been ascribed to BAG3 recently. These previous reports urged us to study the potential involvement of BAG3 in EMT triggered by FGF-2 in renal tubular epithelial cells. The current study found that FGF-2 induced EMT, simultaneously increased BAG3 expression in human kidney 2 (HK2) cells. Although FGF-2 induced EMT in nontransfected or scramble short hairpin RNA (shRNA) transfected HK2 cells, it was ineffective in BAG3-silenced cells, indicating a favorable role of BAG3 in EMT of tubular cells induced by FGF-2. Knockdown of BAG3 also significantly suppressed motion and invasion of HK2 cells mediated by FGF-2. Furthermore, we confirmed that BAG3 was upregulated in kidney of unilateral ureteral obstruction (UUO) rats, a well-established renal fibrosis model, in which EMT is supposed to exert a substantial influence on renal fibrosis. Importantly, upregulation of BAG3 was limited to tubular epithelial cells. Results of the current study identify BAG3 as a potential player in EMT of tubular epithelial cells, as well as renal fibrosis. 相似文献
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The epithelial-mesenchymal transition (EMT) plays a crucial role in the differentiation of many tissues and organs. So far, an EMT was not detected in the development of the auditory organ. To determine whether an EMT may play a role in the morphogenesis of the auditory organ, we studied the spatial localization of several EMT markers, the cell-cell adhesion molecules and intermediate filament cytoskeletal proteins, in epithelium of the dorsal cochlea during development of the rat Corti organ from E18 (18th embryonic day) until P25 (25th postnatal day). We examined by confocal microscopy immunolabelings on cryosections of whole cochleae with antibodies anti-cytokeratins as well as with antibodies anti-vimentin, anti-E-cadherin and anti-β-catenin. Our results showed a partial loss of E-cadherin and β-catenin and a temporary appearance of vimentin in pillar cells and Deiters between P8 and P10. These observations suggest that a partial EMT might be involved in the remodelling of the Corti organ during the postnatal stages of development in rat. 相似文献
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Recently a number of computational approaches have been developed for the prediction of protein–protein interactions. Complete
genome sequencing projects have provided the vast amount of information needed for these analyses. These methods utilize the
structural, genomic, and biological context of proteins and genes in complete genomes to predict protein interaction networks
and functional linkages between proteins. Given that experimental techniques remain expensive, time-consuming, and labor-intensive,
these methods represent an important advance in proteomics. Some of these approaches utilize sequence data alone to predict
interactions, while others combine multiple computational and experimental datasets to accurately build protein interaction
maps for complete genomes. These methods represent a complementary approach to current high-throughput projects whose aim
is to delineate protein interaction maps in complete genomes. We will describe a number of computational protocols for protein
interaction prediction based on the structural, genomic, and biological context of proteins in complete genomes, and detail
methods for protein interaction network visualization and analysis. 相似文献
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Ayhan Ünlü 《Molecular biology reports》2014,41(1):355-364
Actin is one of the most abundant proteins in eukaryotic cells, where it plays key roles in cell shape, motility, and regulation. Actin is found in globular (G) and filamentous (F) structure in the cell. The helix of actin occurs as a result of polymerization of monomeric G-actin molecules through sequential rowing, is called F-actin. Recently, the crystal structure of an actin dimer has been reported, which details molecular interface in F-actin. In this study, the computational prediction model of actin and actin complex has been constructed base on the atomic model structure of G-actin. To this end, a docking simulation was carried out using predictive docking tools to obtain modeled structures of the actin–actin complex. Following molecular dynamics refinement, hot spots interactions at the protein interface were identified, that were predicted to contribute substantially to the free energy of binding. These provided a detailed prediction of key amino acid interactions at the protein–protein interface. The obtained model can be used for future experimental and computational studies to draw biological and functional conclusions. Also, the identified interactions will be used for designing next studies to understand the occurrence of F-actin structure. 相似文献
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Yamada M Kuwano K Maeyama T Hamada N Yoshimi M Nakanishi Y Kasper M 《Histochemistry and cell biology》2008,129(4):453-462
epithelial–mesenchymal transition (EMT) has been considered to be involved in organ fibrogenesis. However, there is few direct
evidence of this process in the pathophysiology of pulmonary fibrosis in vivo. Therefore, we tried to verify the involvement
of this process in the development of pulmonary fibrosis. Since the co-expressions of epithelial and mesenchymal markers are
thought to be a marker of EMT, we performed dual-immuunohistochemistry to assess the co-expressions of these proteins in lung
tissues from bleomycin-induced pulmonary fibrosis in mice, and from patients with idiopathic pulmonary fibrosis, and nonspecific
interstitial pneumonia. Double positive cells for epithelial markers including E-cadherin, T1α, or aquaporin 5, and a mesenchymal
markers α-smooth muscle actin or vimentin were not found in bleomycin-induced pulmonary fibrosis in mice. Double positive
cells for E-cadherin, ICAM-1, LEA, CD44v9, or SP-A and α-smooth muscle actin or vimentin were not found in lung tissues from
normal lung parenchyma, idiopathic pulmonary fibrosis and nonspecific interstitial pneumonia. These results offer at least
two possibilities. One is that EMT does not occur in IPF or bleomycin-induced pulmonary fibrosis in mice. Another is that
EMT may occur in pulmonary fibrosis but the time during this transition in which cells express detectable levels of epithelial
and mesenchymal markers is too small to be detected by double immunohistochemistry. 相似文献