1.Analysis of Proteoglycan Gene Messages in Human Nasal Mucosa and Nasal Polyp using Dot Blot Hybridization.
Ji Hoon PARK ; Seung Kuk BAEK ; Chan Ki YOO ; Jeong Joon KIM ; Sung Dong CHO ; Heung Man LEE ; Sang Hag LEE
Korean Journal of Otolaryngology - Head and Neck Surgery 2001;44(6):606-610
BACKGROUND AND OBJECTIVES: It has been suggested that the formation and growth of nasal polyp require the remodeling of extracellular matrix. Proteoglycans (PGs) are the major components of the extracellular matrix that maintain the integrity of the structural tissues The leucine-rich repeat PGs include lumican, decorin and biglycan, all of which have many important biologic activities in various pathologic conditions, including the remodeling of the extracellular matrix. Therefore, these small-PG families may be involved in the formation and growth of nasal polyp. MATERIALS AND METHODS: Surgical specimens of nasal polyps and the normal nasal mucosa were assessed for mRNA expressions coding for lumican, decorin and biglycan using the reverse transcription-polymerase chain reaction,which was followed by dot blot hybridization. RESULTS: Lumican, decorin and biglycan mRNAs were expressed in all tissue samples examined. Semi-quantitative dot blot hybridization revealed that the levels of the lumican and biglycan messages are lower in the nasal polyp tissues than in the nasal mucosa. The decorin messages in the nasal polyp were expressed at levels similar to those in the nasal mucosa. CONCLUSION: These results suggest that lumican, decorin and biglycan may be important components of the extracellular matrix in the nasal mucosa. Considering the function of these PGs, the normal levels of decorin associated with low levels of biglycan and lumican may play a role in the pathogenesis of nasal polyposis.
Biglycan
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Clinical Coding
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Decorin
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Extracellular Matrix
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Humans*
;
Nasal Mucosa*
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Nasal Polyps*
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Proteoglycans*
;
RNA, Messenger
2.Histopathological Characteristics of Human Coronary Stent Restenosis.
Korean Circulation Journal 2000;30(1):5-15
BACKGROUND AND OBJECTIVES: Neointimal ingrowth rather than stent recoil is thought to be important for coronary in-stent restenosis. However only limited pathologic data are available to adress the mechanisms of in-stent restenosis. With the specific aim of measuring cell replication and of assessing cellularity and extracellular matrix (ECM) composition, we analyzed atherectomized coronary arterial in-stent restenotic specimens. METHODS AND RESULTS: In the present study, we analyzed 29 atherectomized coronary arterial in-stent restenotic tissue samples (14 LAD, 10 RCA, and 5 LCX) retrieved from 25 patients (m/f:18/7: age 59+/-13 yr) at 0.5-23 (mean 5.7) months after deployment of Palmaz-Schatz stent. Histopathological analysis of cellular components and ECM was performed using H & E, modified Movat pentachrome, and immunocytochemical staining. Cellular proliferation rate, as estimated by use of antibodies to Ki-67 nuclear antigen showed low proliferation rate with the range of 0-4%, and no positive cells were found in 62% of cases. Myxoid tissue having ECM enriched with versican and hyaluronan was found in 69% of cases, and decreased over time after stenting. Foci of cell poor area were found in 57% of cases, and could be classified into as: (1) containing collagen-rich ECM and (2) containing a proteoglycan-rich ECM. Versican, biglycan, perlecan, and hyaluronan were identified with varying individual distributions in the proteoglycan rich area. Specimens with foci of cell poor area tended to increase over time after stenting (31% in & 4 mo vs. 81% in > or =4 mo after stenting, p<0.01). alpha-smooth muscle actin staining identified the majority of cells as smooth muscle cells (SMC) and occasional macrophages (< or =12 cells per section) were detected by CD68 antibody. CONCLUSIONS: These data suggest that enhanced ECM accumulation rather than cell proliferation may be important mechanisms for stent restenosis. Angioplasty of stent restenosis may therefore fail due to transient compression of this hygroscopic matrix.
Actins
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Angioplasty
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Antibodies
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Biglycan
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Cell Proliferation
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Extracellular Matrix
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Humans*
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Hyaluronic Acid
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Macrophages
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Myocytes, Smooth Muscle
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Proteoglycans
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Stents*
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Versicans
3.Immunohistochemical localization of leucine-rich proteoglycans in the developing periodontal tissues of mice.
Shu LI ; Pi-shan YANG ; Ke-qing PAN ; Rui-yue XIE ; Xue-jing DUAN
Chinese Journal of Stomatology 2005;40(6):485-488
OBJECTIVETo study the distribution and expression of fibromodulin, decorin and biglycan in developing normal periodontal tissues, so as to understand its role in periodontal tissue formation.
METHODSThirty six BALB/c mice in different developing stages were killed and their bilateral mandibular first molars with surrounding alveolar bones and gingival tissues were taken out, Power Vision two steps immunohistochemical method with anti-fibromodulin, anti-decorin and anti-biglycan was used to detect the tissue distribution and cellular localization of fibromodulin and related proteoglycans, decorin and biglycan.
RESULTSFibromodulin was strongly expressed in the subcutaneous gingival connective tissue, periodontal ligament, mainly in gingival and periodontal fibroblasts as well as their matrices. Strong expression was also noted in the area close to the interfaces of periodontal ligament-alveolar bone and periodontal ligament-cementum. Decorin was strongly expressed in the area of gingival connective tissue, periodontal ligament and the surface of alveolar bone, while biglycan was stained evidently in gingival connective tissue throughout the period of investigation, but negative in the surface of alveolar bone and osteoblasts.
CONCLUSIONSFibromodulin may interact with decorin and biglycan to regulate the network formation of gingival connective tissues and periodontal collagen fibers, and may be involved in mineralization of the alveolar bone and cementum.
Alveolar Process ; cytology ; growth & development ; Animals ; Biglycan ; Decorin ; Extracellular Matrix Proteins ; analysis ; Fibromodulin ; Gingiva ; chemistry ; growth & development ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Osteoblasts ; chemistry ; Periodontal Ligament ; chemistry ; growth & development ; Proteoglycans ; analysis ; Tooth Germ ; chemistry
4.Signaling of biglycan and cytokines in bovine intervertebral disc cells.
Journal of Central South University(Medical Sciences) 2011;36(9):889-894
OBJECTIVE:
To evaluate the effect of biglycan on the signaling of cytokines (epidermal growth factor, osteogenic protein-1, and interleukin-1) in bovine intervertebral disc cells.
METHODS:
Nucleoplasty (NP) and annulus fibrosus (AF) cells of the intervertebral disc tissues were isolated from the tails of young adult bovine. First, the cells were treated in 3 ways: Biglycan alone, cytokines alone (epidermal growth factor, osteogenic protein-1, or interleukin-1), and biglycan combined with cytonkines. Western blot was used to observe the singling of biglycan and cytokines in bovine intervertebral disc cells, and to identify the effect of biglycan on cytokines mentioned above.
RESULTS:
Biglycan upregulated the signaling (3- 4 folds) with the optimal effect at 10 min and 20 μmol/L both in the AF cells and NP cells. Epidermal growth factor, osteogenic protein-1, or interleukin-1 also upregulated the protein expression in the extracellular matrix of intervertebral disc cells. When combined different biglycan concentrations with epidermal growth factor, osteogenic protein-1, or interleukin-1 to treat the intervertebral disc cells, the concentration of biglycan rose, whereas the cytokine signal decreased both in the bovine AF and NP cells (P<0.01). There was no significant difference between the AF and NP cells.
CONCLUSION
Biglycan can adhere to the intervertebral disc cells to activate the extracellular signal-regulated kinase (ERK) pathway and this effect is time and concentration dependent. Byglycan can decrease not only the anabolism effect of epidermal growth factor and osteogenic protein-1, but also the catabolism effect of interleukin-1. This regulatory role of biglycan may be very important to maintain the metabolism balance. Biglycan may be good for the repair of intervertebral disc.
Animals
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Biglycan
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physiology
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Bone Morphogenetic Protein 7
;
metabolism
;
physiology
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Cattle
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Cells, Cultured
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Cytokines
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metabolism
;
physiology
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Epidermal Growth Factor
;
metabolism
;
physiology
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Interleukin-1
;
metabolism
;
physiology
;
Intervertebral Disc
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cytology
;
metabolism
;
Signal Transduction
5.Gene expression profiling of diffuse-type gastric cancer by cDNA microarray.
Xiao-qing ZHANG ; Yan-qing YANG ; Bing-ya LIU ; Xiao-long JIN ; Wei LI ; Kai-ling TANG ; Qing-hua ZHANG ; Yan-zhen LIN ; Zheng-gang ZHU
Chinese Journal of Oncology 2006;28(2):116-119
OBJECTIVETo identify cancer-related genes in diffuse-type gastric cancer and to explore its molecular mechanism by cDNA microarray analysis.
METHODSA total of 22 pairs of diffuse-type gastric cancer tissue and the corresponding normal mucosa were taken and freshly frozen. cDNA microarray with 14,592 genes/ESTs was used. Genes were considered to be up- or down-regulated when the fluorescent intensity ratio between tumor and normal mucosa was over 2-fold in over 50% of the samples (P < 0.05). Hierarchical clustering of regulated genes was performed as a measure to study expressional similarity. Validation of array results was carried out by real time quantitative PCR (QPCR).
RESULTSCompared with those of corresponding normal mucosa, there were a total of 153 genes/ESTs up-regulated and 204 down-regulated in diffuse-type gastric cancer. Hierarchical clustering demonstrated that the genes belonging to the same subgroup displayed similar function. Most of the overexpressed genes were those related to cell adhesion, cell motility, matrix reconstruction, cell proliferation and/or signal transduction; while genes related to defense response, toxicoid metabolism, DNA repairing, nuclear-cytoplasmic transport and/or anti-apoptosis made up the main list of the underexpressed genes. Seven genes showed higher expression in TNM (T I + T II) group than in (T III + T IV) group. QPCR confirmed the array analysis results.
CONCLUSIONGene expression profiling by cDNA microarray analysis provides not only molecular understanding of biological properties of cancer, but may also be helpful in discovering new diagnostic markers and therapeutic targets in gastric adenocarcinoma.
Adenocarcinoma ; genetics ; metabolism ; Biglycan ; Collagen Type I ; metabolism ; Expressed Sequence Tags ; Extracellular Matrix Proteins ; metabolism ; Female ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Microfilament Proteins ; metabolism ; Middle Aged ; Muscle Proteins ; metabolism ; Neoplasm Staging ; Oligonucleotide Array Sequence Analysis ; Pepsinogen C ; metabolism ; Proteoglycans ; metabolism ; Stomach Neoplasms ; genetics ; metabolism
6.Microarray Analysis of Gene Expression During Differentiation of Human Mesenchymal Stem Cells Treated with Vitamin E in vitro into Osteoblasts.
Ki Hoon AHN ; Hwa Kyung JUNG ; So Eun JUNG ; Kyong Wook YI ; Hyun Tae PARK ; Jung Ho SHIN ; Young Tae KIM ; Jun Young HUR ; Sun Haeng KIM ; Tak KIM
Korean Journal of Bone Metabolism 2011;18(1):23-32
OBJECTIVE: Supplementation with vitamin E is able to protect bone against free radical-induced elevation of bone-resorbing cytokines. We examined gene expression by microarray analysis during the differentiation of human mesenchymal stem cells treated with vitamin E into osteoblasts in vitro. METHODS: Human bone marrow stem cells were cultured in osteogenic differentiation medium and vitamin E was added. A colorimetric immunoassay for the quantification of cell proliferation was used to measure osteoblast differentiation. Gene expression was analyzed using a microarray technique. We also used a real time reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: It was found that vitamin E enhanced cell proliferation when compared to cells cultured in media without vitamin E. We focused on 68 genes which are related to osteogenesis and osteoclastogenesis. Alkaline phosphatase, transforming growth factor-beta 1, fibroblast growth factor receptor 1, matrix metalloproteinase 2, muscle segment homeobox 2, bone morphogenetic protein 1, biglycan, vascular endothelial growth factor B, dentin sialophosphoprotein, cartilage oligomeric matrix protein, runt-related transcription factor 2, fibroblast growth factor receptor 3, and SMAD2 were upregulated > 2-fold compared to the control. Conversely, osteopetrosis-associated transmembrane protein 1, microphthalmia-associated transcription factor, and epidermal growth factor receptor were downregulated > 2-fold compared to the control. Vitamin E produced a 1.5-fold increase in the expression of runt-related transcription factor 2 and transforming growth factor-beta 1 as determined by real time RT-PCR. CONCLUSION: Vitamin E had a positive effect on the gene expressions regarding osteogenic differentiation of mesenchymal stem cells.
Alkaline Phosphatase
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Biglycan
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Bone Marrow
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Bone Morphogenetic Protein 1
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Cartilage
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Cell Proliferation
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Cytokines
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Dentin
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Durapatite
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Extracellular Matrix Proteins
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Gene Expression
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Genes, Homeobox
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Glycoproteins
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Humans
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Immunoassay
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Matrix Metalloproteinase 2
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Mesenchymal Stromal Cells
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Microarray Analysis
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Microphthalmia-Associated Transcription Factor
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Muscles
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Osteoblasts
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Osteogenesis
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Phosphoproteins
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Receptor, Epidermal Growth Factor
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Receptor, Fibroblast Growth Factor, Type 1
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Receptor, Fibroblast Growth Factor, Type 3
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Sialoglycoproteins
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Stem Cells
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Transcription Factors
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Vascular Endothelial Growth Factor B
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Vitamin E
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Vitamins