Title: miR-9/MCPIP1 axis mediated regulation of IL-6 expression in osteoarthritis chondrocytes
3rd International Conference and Exhibition on Orthopedics & Rheumatology, July 28-30, 2014, San Francisco, USA
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Abstract
Background/Purpose: Post-transcriptional regulation of cytokine expression is important for maintaining tissue integrity. MCPIP1 was identified as a novel protein, which destabilizes inflammatory cytokines mRNAs via their 3’ UTR. IL-6 has recently gained attention because of its high levels in synovial fluid in Osteoarthritis (OA) and ability to induce high levels of MMP-13 in OA. In the present study we determined whether MCPIP1 regulates IL-6 expression and evaluated the role of miR- 9/MCPIP1 axis in the regulation of IL-6 in human OA chondrocytes.
Methods: Human chondrocytes were prepared from OA cartilage by the enzymatic digestion. TaqMan assays were used for gene expression analysis using RNA isolated from cultured primary chondrocytes or from damaged or smooth regions of OA cartilage or RNA immunoprecipitation (RIP). RNA fluorescent in-situ hybridization (ISH) for IL-6 and MCPIP1 expression was performed using RNAScope. Transfection was done using Amaxa kit. Knockdown experiments were performed using Trisilencer-27 human siRNA. For RIP, lysates from IL-1β-stimulated chondrocytes were incubated overnight with anti- MCPIP1 antibody or with isotype control IgG followed by RNA purification.
Biography
Tariq M Haqqi completed his PhD from Aligarh Muslim University, India and postdoctoral studies from Marseille-Luminy, France and Mayo Clinic, Rochester, MN. He is Professor of Anatomy and Neurobiology at the Northeast Ohio Medical University, Rootstown, OH. He has published over 80 papers in highly reputed journals and has served as Editor-in-Chief and Editorial Board Member of reputed journals.
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