Transcriptional profiling of bovine intervertebral disc cells implications for identification of normal and degenerate human intervertebral disc cell phenotypes 英文参考文献.docVIP

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Transcriptional profiling of bovine intervertebral disc cells implications for identification of normal and degenerate human intervertebral disc cell phenotypes 英文参考文献.doc

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Transcriptional profiling of bovine intervertebral disc cells implications for identification of normal and degenerate human intervertebral disc cell phenotypes 英文参考文献

Minogue et al. Arthritis Research Therapy 2010, 12:R22 /content/12/1/R22 RESEARCH ARTICLE Open Access Transcriptional profiling of bovine intervertebral Research article disc cells: implications for identification of normal and degenerate human intervertebral disc cell phenotypes Ben M Minogue1, Stephen M Richardson1, Leo AH Zeef2, Anthony J Freemont1 and Judith A Hoyland*1 Abstract Introduction: Nucleus pulposus (NP) cells have a phenotype similar to articular cartilage (AC) cells. However, the matrix of the NP is clearly different to that of AC suggesting that specific cell phenotypes exist. The aim of this study was to identify novel genes that could be used to distinguish bovine NP cells from AC and annulus fibrosus (AF) cells, and to further determine their expression in normal and degenerate human intervertebral disc (IVD) cells. Methods: Microarrays were conducted on bovine AC, AF and NP cells, using Affymetrix Genechip? Bovine Genome Arrays. Differential expression levels for a number of genes were confirmed by quantitative real time polymerase chain reaction (qRT-PCR) on bovine, AC, AF and NP cells, as well as separated bovine NP and notochordal (NC) cells. Expression of these novel markers were further tested on normal human AC, AF and NP cells, and degenerate AF and NP cells. Results: Microarray comparisons between NP/ACAF and NP/AC identified 34 NP-specific and 49 IVD-specific genes respectively that were differentially expressed ≥100 fold. A subset of these were verified by qRT-PCR and shown to be expressed in bovine NC cells. Eleven genes (SNAP25, KRT8, KRT18, KRT19, CDH2, IBSP, VCAN, TNMD, BASP1, FOXF1 FBLN1) were also differentially expressed in normal human NP cells, although to a lesser degree. Four genes (SNAP25, KRT8, KRT18 and CDH2) were significantly decreased in degenerate human NP cells, while three genes (VCAN, TNMD and BASP1) were significantly increased in degenerate human AF cells. The IVD negative marker FBLN1 was