title: [Analysis of gene expression patterns in rheumatoid synovial fibroblasts using RAP-PCR for differential display].
abstract: OBJECTIVE
  Destruction of articular cartilage and bone by invading synovial fibroblasts is a typical histopathologic feature in rheumatoid arthritis (RA). However, little is known about specific up- or downregulation of genes leading to this aggressive phenotype. Thus, our aim was to identify genes, which are differentially expressed in RA synovial fibroblasts as compared to synovial fibroblasts derived from patients with osteoarthritis (OA) using RAP-PCR for differential display.
  
  
  METHODS
  After extraction of total RNA, the first step of RAP-PCR was performed using various different arbitrary 10-12-base primers for first-strand cDNA synthesis. Second-strand synthesis was achieved by cycling at low stringency conditions for 35 cycles using different arbitrary 10-base primers, followed by electrophoretic separation and sequence analysis of the amplified fingerprint products.
  
  
  RESULTS
  On average, approximately 70 different RNAs were obtained per primer, of which most were expressed both by RA and OA synovial fibroblasts. Using 26 different primer combinations, in total 12 cDNAs were differentially expressed between RA and OA synovial fibroblasts. In the RA group strong amplification of distinct PCR products suitable for sequencing could be observed. Sequence analysis identified these PCR products as highly homologous to various genes involved in regulation of cell cycle and metabolism.
  
  
  CONCLUSIONS
  The data indicate that RAP-PCR is a suitable method to identify differentially expressed genes in rheumatoid synovial fibroblasts potentially involved in the specific pathophysiology of RA.
authors.affiliation: Klinik und Poliklinik für Innere Medizin I, Universität Regensburg. ulf.mueller-ladner@klinik.uni-regensburg.de
authors.forename: U
authors.initials: U
authors.lastname: Müller-Ladner
authors.forename: M
authors.initials: M
authors.lastname: Judex
authors.forename: H P
authors.initials: HP
authors.lastname: Jüsten
authors.forename: D
authors.initials: D
authors.lastname: Wessinghage
authors.forename: J
authors.initials: J
authors.lastname: Welsh
authors.forename: M
authors.initials: M
authors.lastname: McClelland
authors.forename: S
authors.initials: S
authors.lastname: Gay
authors.forename: J
authors.initials: J
authors.lastname: Schölmerich
authors.forename: F
authors.initials: F
authors.lastname: Kullmann
mesh_terms: D001172:Arthritis, Rheumatoid / Q000235:genetics*; D005347:Fibroblasts / Q000737:chemistry / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012313:RNA / Q000032:analysis; D013583:Synovial Membrane / Q000737:chemistry / Q000378:metabolism
chemical_list: D012313:RNA
vernacular_title: Analyse des Genexpressionsmusters von synovialen Fibroblasten von Patienten mit rheumatoider Arthritis mittels RAP-PCR Differential Display.