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Molecular characterization and partial cDNA cloning of facilitative glucose transporters expressed in human articular chondrocytes; stimulation of 2-deoxyglucose uptake by IGF-I and elevated MMP-2 secretion by glucose deprivation.

Richardson, S, Neama, G, Phillips, T, Bell, S, Carter, SD, Moley, KH, Moley, JF, Vannucci, SJ and Mobasheri, A (2003) Molecular characterization and partial cDNA cloning of facilitative glucose transporters expressed in human articular chondrocytes; stimulation of 2-deoxyglucose uptake by IGF-I and elevated MMP-2 secretion by glucose deprivation. Osteoarthritis Cartilage, 11 (2). pp. 92-101.

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Abstract

OBJECTIVE: Recent evidence suggests that human chondrocytes express several facilitative glucose transporter (GLUT) isoforms and also that 2-deoxyglucose transport is accelerated by cytokine stimulation. The aim of the present investigation was to determine if human articular chondrocytes express any of the recently identified members of the GLUT/SLC2A gene family and to examine the effects of endocrine factors, such as insulin and IGF-I on the capacity of human chondrocytes for transporting 2-deoxyglucose. DESIGN/METHODS: PCR, cloning and immunohistochemistry were employed to study the expression of GLUT/SLC2A transporters in normal human articular cartilage. The uptake of 2-deoxyglucose was examined in monolayer cultured immortalized human chondrocytes following stimulation with TNF-alpha, insulin and IGF-I. Levels of MMP-2 were assessed by gelatin zymography following glucose deprivation of alginate cultures. RESULTS: Using PCR we detected transcripts for eight glucose transporter isoforms (GLUTs 1, 3, 6, 8, 9, 10, 11 and 12) and for a fructose transporter (GLUT5) in human articular cartilage. Expression of GLUT1, GLUT3 and GLUT9 proteins in normal human articular cartilage was confirmed by immunohistochemistry. The uptake of 2-deoxyglucose was dependent on time and temperature, inhibited by cytochalasin B and phloretin, and significantly accelerated in chondrocyte cultures stimulated with IGF-I. However, 2-deoxyglucose uptake was unaffected by short and long-term insulin treatment, which ruled out a functional role for insulin-sensitive GLUT4-mediated glucose transport. Furthermore, secretion of MMP-2 was increased in alginate cultures deprived of glucose. CONCLUSIONS: The data supports a critical role for glucose transport and metabolism in the synthesis and degradation of cartilage.

Item Type: Article
Authors :
NameEmailORCID
Richardson, SUNSPECIFIEDUNSPECIFIED
Neama, GUNSPECIFIEDUNSPECIFIED
Phillips, TUNSPECIFIEDUNSPECIFIED
Bell, SUNSPECIFIEDUNSPECIFIED
Carter, SDUNSPECIFIEDUNSPECIFIED
Moley, KHUNSPECIFIEDUNSPECIFIED
Moley, JFUNSPECIFIEDUNSPECIFIED
Vannucci, SJUNSPECIFIEDUNSPECIFIED
Mobasheri, Aa.mobasheri@surrey.ac.ukUNSPECIFIED
Date : February 2003
Uncontrolled Keywords : Blotting, Western, Cartilage, Articular, Cell Line, Chondrocytes, Cytochalasin B, DNA, Circular, Deoxyglucose, Glucose, Growth Substances, Hot Temperature, Humans, Immunohistochemistry, Insulin, Intercellular Signaling Peptides and Proteins, Matrix Metalloproteinase 2, Monosaccharide Transport Proteins, Phloretin, Polymerase Chain Reaction, Protein Isoforms, Time Factors, Tumor Necrosis Factor-alpha
Related URLs :
Depositing User : Symplectic Elements
Date Deposited : 17 May 2017 10:16
Last Modified : 17 May 2017 14:48
URI: http://epubs.surrey.ac.uk/id/eprint/827139

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