Abstract
To select candidate genes, we attempted to comparative analysis of protein levels between rheumatoid arthritis (RA) patients and healthy controls by two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF-MS). We identified 17 proteins that showed up- or down-regulated spots in RA patients. We found that coactosin-like1 (COTL1) were highly expressed in RA patients compared with healthy controls. We performed a case-control study to determine whether the COTL1 gene polymorphisms were associated with RA and systemic lupus erythematosus (SLE). The genotype frequency of c.-1124G>T and the allelic frequency of c.484G>A in RA patients, and the genotype frequency of c.484G>A in SLE patients were significantly different from healthy controls (P=0.009, 0.027, and 0.025, respectively). We also investigated the correlation with the levels of rheumatoid factor (RF) and anti-cyclic citrullinated peptide (CCP) antibody in RA patients, and anti-nuclear antibodies (ANA) in SLE patients. The c.484G>A polymorphism in RA patients has significant association with the levels of anti-CCP antibody (P=0.03). Our findings demonstrated that c.-1124G>T and c. 484G>A polymorphisms of the COTL1 gene might be associated with the genetic susceptibility of autoimmune disorders.
Highlights
Rheumatoid arthritis (RA) and systematic lupus erythematosus (SLE) are the complex autoimmune diseases which are caused by combination of multiple genetic, hormonal, and environmental factors (Gregersen, 1999; Kotzin, 1996)
We identified COTL1 protein which is highly expressed in RA patients by comparative proteome analysis, and compared the mRNA expression level of COTL1 between RA patients and healthy controls by RT-PCR
We further investigated the relationship between genotypes of each polymorphism and rheumatoid factor (RF) or anti-cyclic citrullinated peptide (CCP) antibody levels in RA patients, and antinuclear antibody (ANA) levels in SLE patients
Summary
Rheumatoid arthritis (RA) and systematic lupus erythematosus (SLE) are the complex autoimmune diseases which are caused by combination of multiple genetic, hormonal, and environmental factors (Gregersen, 1999; Kotzin, 1996). RFs and anti-CCP antibody are used in clinical practice (Bläss et al, 1999, 2001; Schellekens et al, 2000) They can be detactable up to 70-80% in RA patients (Mewar et al, 2006; Raptopoulou et al, 2007). Two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) have become a powerful method for comparative proteome analysis (O'Farrell, 1975; Wang et al, 1994; Mann et al, 1996; Lottspeich, 1999). These techniques have been used for studying RA using the samples derived from RA patients such as plasma, synovial fluid, and peripheral blood mononuclear cells. We further investigated the relationship between genotypes of each polymorphism and RF or anti-CCP antibody levels in RA patients, and ANA levels in SLE patients
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