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1.
Eur Rev Med Pharmacol Sci ; 22(17): 5481-5489, 2018 09.
Article in English | MEDLINE | ID: mdl-30229819

ABSTRACT

OBJECTIVE: Desmoglein-2 (Dsg2) plays a crucial role in the assembly and adhesion of desmosomes. The absent or aberrant expression of Dsg2 was reported to be associated with the progression of varies human cancers. However, the expression of Dsg2 in hepatocellular carcinoma (HCC) and its association with tumor prognosis is still unknown. The aim of this study was to evaluate the expression level of Dsg2 in HCC and of the correlation between Dsg2 expression and clinicopathological variables. PATIENTS AND METHODS: A total of 104 patients diagnosed with HCC were enrolled in this study. Real time-quantitative PCR (RT-qPCR) and Western blot were performed to determine the expression level of Dsg2 in HCC tumor tissues and matched noncancerous tissues. Cell proliferation and cell cycle were measured by cell counting kit-8 (CCK-8) assay and flow-cytometry assay, respectively. RESULTS: Our results revealed that Dsg2 expression was significantly higher in HCC tumor tissues than in matched noncancerous tissues (p < 0.01), positively correlated with tumor size (p = 0.035) and tumor stage (p = 0.021). Univariate and multivariate analyses demonstrated Dsg2 expression was an independent prognostic factor for overall survival. Meanwhile, we found knockdown the expression of Dsg2 using small interfering RNA (siRNA) could efficiently impaired HCC cell proliferation rate and cell cycle progression (p < 0.05). CONCLUSIONS: Taken together, our results suggest that increased Dsg2 expression was associated with tumor progression in HCC and may function as a promising biomarker for unfavorable prognosis of HCC.


Subject(s)
Carcinoma, Hepatocellular/diagnosis , Carcinoma, Hepatocellular/metabolism , Desmoglein 2/biosynthesis , Liver Neoplasms/diagnosis , Liver Neoplasms/metabolism , Adult , Biomarkers, Tumor/biosynthesis , Biomarkers, Tumor/genetics , Carcinoma, Hepatocellular/genetics , Cell Line, Tumor , Cell Proliferation/genetics , Desmoglein 2/genetics , Female , Gene Expression Regulation, Neoplastic , Humans , Liver Neoplasms/genetics , Male , Middle Aged , Prognosis
2.
Am J Physiol Gastrointest Liver Physiol ; 313(5): G478-G491, 2017 Nov 01.
Article in English | MEDLINE | ID: mdl-28864500

ABSTRACT

Krüppel-like factor 5 (KLF5) is a member of the zinc finger family of transcription factors that regulates homeostasis of the intestinal epithelium. Previous studies suggested an indispensable role of KLF5 in maintaining intestinal barrier function. In the current study, we investigated the mechanisms by which KLF5 regulates colonic barrier function in vivo and in vitro. We used an inducible and a constitutive intestine-specific Klf5 knockout mouse models (Villin-CreERT2;Klf5fl/fl designated as Klf5ΔIND and Villin-Cre;Klf5fl/fl as Klf5ΔIS ) and studied an inducible KLF5 knockdown in Caco-2 BBe cells using a lentiviral Tet-on system (Caco-2 BBe KLF5ΔIND). Specific knockout of Klf5 in colonic tissues, either inducible or constitutive, resulted in increased intestinal permeability. The phenotype was accompanied by a significant reduction in Dsg2, which encodes desmoglein-2, a desmosomal cadherin, at both mRNA and protein levels. Transmission electron microscopy showed alterations of desmosomal morphology in both KLF5 knockdown Caco-2 BBe cells and Klf5 knockout mouse colonic tissues. Inducible knockdown of KLF5 in Caco-2BBe cells grown on Transwell plates led to impaired barrier function as evidenced by decreased transepithelial electrical resistance and increased paracellular permeability to fluorescein isothiocyanate-4 kDa dextran. Furthermore, DSG2 was significantly decreased in KLF5 knockdown cells, and DSG2 overexpression partially rescued the impaired barrier function caused by KLF5 knockdown. Electron microscopy studies demonstrated altered desmosomal morphology after KLF5 knockdown. In combination with chromatin immunoprecipitation analysis and promoter study, our data show that KLF5 regulates intestinal barrier function by mediating the transcription of DSG2, a gene encoding a major component of desmosome structures.NEW & NOTEWORTHY The study is original research on the direct function of a Krüppel-like factor on intestinal barrier function, which is commonly exerted by cell junctions, including tight junctions, adherens junctions, and desmosomes. Numerous previous studies were focused on tight junctions and adherens junctions. However, this study provided a new perspective on how the intestinal barrier function is regulated by KLF5 through DSG2, a component of desmosome complexes.


Subject(s)
Colon/physiology , Kruppel-Like Transcription Factors/physiology , Animals , Caco-2 Cells , Desmocollins , Desmoglein 2/biosynthesis , Desmoglein 2/genetics , Desmosomes/ultrastructure , Electric Impedance , Gene Expression Regulation/genetics , Gene Knockdown Techniques , HEK293 Cells , Humans , Intestinal Mucosa/metabolism , Intestinal Mucosa/ultrastructure , Membrane Glycoproteins/biosynthesis , Membrane Glycoproteins/genetics , Mice , Mice, Inbred C57BL , Permeability , RNA, Messenger/biosynthesis , RNA, Messenger/genetics
3.
Oncotarget ; 7(25): 37536-37555, 2016 Jun 21.
Article in English | MEDLINE | ID: mdl-26918609

ABSTRACT

The desmosomal cadherin, desmoglein 2 (Dsg2), is deregulated in a variety of human cancers including those of the skin. When ectopically expressed in the epidermis of transgenic mice, Dsg2 activates multiple mitogenic signaling pathways and increases susceptibility to tumorigenesis. However, the molecular mechanism responsible for Dsg2-mediated cellular signaling is poorly understood. Here we show overexpression as well as co-localization of Dsg2 and EGFR in cutaneous SCCs in vivo. Using HaCaT keratinocytes, knockdown of Dsg2 decreases EGFR expression and abrogates the activation of EGFR, c-Src and Stat3, but not Erk1/2 or Akt, in response to EGF ligand stimulation. To determine whether Dsg2 mediates signaling through lipid microdomains, sucrose density fractionation illustrated that Dsg2 is recruited to and displaces Cav1, EGFR and c-Src from light density lipid raft fractions. STED imaging confirmed that the presence of Dsg2 disperses Cav1 from the cell-cell borders. Perturbation of lipid rafts with the cholesterol-chelating agent MßCD also shifts Cav1, c-Src and EGFR out of the rafts and activates signaling pathways. Functionally, overexpression of Dsg2 in human SCC A431 cells enhances EGFR activation and increases cell proliferation and migration through a c-Src and EGFR dependent manner. In summary, our data suggest that Dsg2 stimulates cell growth and migration by positively regulating EGFR level and signaling through a c-Src and Cav1-dependent mechanism using lipid rafts as signal modulatory platforms.


Subject(s)
Caveolin 1/metabolism , Desmoglein 2/biosynthesis , ErbB Receptors/biosynthesis , src-Family Kinases/metabolism , CSK Tyrosine-Protein Kinase , Caveolin 1/genetics , Cell Line, Tumor , Cell Movement/physiology , Cell Proliferation/physiology , Desmoglein 2/genetics , Desmoglein 2/metabolism , ErbB Receptors/genetics , ErbB Receptors/metabolism , Fibrosarcoma/genetics , Fibrosarcoma/metabolism , Humans , Membrane Microdomains/enzymology , Membrane Microdomains/metabolism , Signal Transduction , Skin Neoplasms/genetics , Skin Neoplasms/metabolism , Up-Regulation , src-Family Kinases/genetics
4.
Hum Pathol ; 43(10): 1745-54, 2012 Oct.
Article in English | MEDLINE | ID: mdl-22521077

ABSTRACT

Gastroesophageal reflux disease is associated with impaired epithelial barrier function and abnormal expression of proteins forming cell-cell contacts by tight junctions and desmosomes in distal esophageal squamous mucosa. Although gastroesophageal reflux disease and Helicobacter pylori are both associated with chronic inflammation of the adjacent cardia mucosa, it is not known whether these lead to derangements of the desmosomal complexes. Here, we assessed the expression of 4 proteins (plakoglobin and desmoglein 1, 2, and 3) forming epithelial desmosomal complexes by quantitative reverse transcription polymerase chain reaction and immunohistochemistry in biopsies from 67 patients with gastroesophageal reflux disease and 23 gastroesophageal reflux disease-negative controls. Plakoglobin and desmoglein 2 were ubiquitously expressed in all samples, whereas desmoglein 1 and 3 were not expressed in cardia mucosa. Gastroesophageal reflux disease was specifically associated with elevated transcript levels of desmoglein 2 and plakoglobin. These were significantly increased from 2.0- to 2.7-fold in patients with gastroesophageal reflux disease compared with controls (P < .01), and significantly increased immunohistochemical scores for both proteins were observed (P < .05) as well. The combined presence of gastroesophageal reflux disease and Helicobacter pylori infection had no additional effect on desmosomal gene expression. Taken together, the up-regulation of plakoglobin and desmoglein 2 in cardia mucosa of patients with gastroesophageal reflux disease supports the concept that the "transition zone" between distal esophagus and proximal stomach is affected by gastroesophageal reflux disease as well, and architectural and molecular changes in the desmosomal compartment contribute to the pathogenesis of gastroesophageal reflux disease in the cardia mucosa.


Subject(s)
Desmosomes/metabolism , Gastroesophageal Reflux/metabolism , Gastroesophageal Reflux/microbiology , Helicobacter Infections/metabolism , Adult , Aged , Cardia/metabolism , Cardia/microbiology , Cardia/pathology , Desmoglein 1/analysis , Desmoglein 1/biosynthesis , Desmoglein 2/analysis , Desmoglein 2/biosynthesis , Desmoglein 3/analysis , Desmoglein 3/biosynthesis , Female , Gastric Mucosa/metabolism , Gastric Mucosa/microbiology , Gastric Mucosa/pathology , Helicobacter Infections/complications , Helicobacter Infections/pathology , Helicobacter pylori , Humans , Immunohistochemistry , Male , Middle Aged , Reverse Transcriptase Polymerase Chain Reaction , Up-Regulation , Young Adult , gamma Catenin/analysis , gamma Catenin/biosynthesis
5.
J Invest Dermatol ; 127(9): 2191-206, 2007 Sep.
Article in English | MEDLINE | ID: mdl-17495963

ABSTRACT

During progression of melanomas, a crucial role has been attributed to alterations of cell-cell adhesions, specifically, to a "cadherin switch" from E- to N-cadherin (cad). We have examined the adhesion of melanoma cells to each other and to keratinocytes. When different human melanoma cell lines were studied by protein analysis and immunofluorescence microscopy, six of eight lines contained N-cad, three E-cad, and five P-cad, and some lines had more than one cad. Surprisingly, two N-cad-positive lines, MeWo and C32, also contained desmoglein 2 (Dsg2), a desmosomal cad previously not reported for melanomas, whereas other desmosome-specific proteins were absent. This finding was confirmed by reverse transcriptase-PCR, immunoprecipitation, and matrix-assisted laser desorption ionization-time of flight analyses. Double-label confocal and immunoelectron microscopy showed N-cad, alpha- and beta-catenin in plaque-bearing puncta adhaerentia, whereas Dsg2 was distributed rather diffusely over the cell surface. In cocultures with HaCaT keratinocytes Dsg2 was found in heterotypic cell contact regions. Correspondingly, immunohistochemistry revealed Dsg2 in five of 10 melanoma metastases. Together, we show that melanoma cell adhesions are more heterogeneous than expected and that certain cells devoid of desmosomes contain Dsg2 in a non-junction-restricted form. Future studies will have to clarify the diagnostic and prognostic significance of these different adhesion protein subtypes.


Subject(s)
Desmoglein 2/biosynthesis , Melanoma/metabolism , Melanoma/pathology , Skin Neoplasms/metabolism , Skin Neoplasms/pathology , Cadherins/metabolism , Catenins/metabolism , Cell Adhesion , Cell Communication , Cell Line, Tumor , Desmocollins/metabolism , Desmoglein 2/chemistry , Desmogleins/metabolism , Epidermis/metabolism , Humans , Keratinocytes/metabolism , Microscopy, Fluorescence , Plakophilins/metabolism
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