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01.08.2011 | Research article | Ausgabe 4/2011 Open Access

Arthritis Research & Therapy 4/2011

Investigating a pathogenic role for TXNDC5 in rheumatoid arthritis

Zeitschrift:
Arthritis Research & Therapy > Ausgabe 4/2011
Autoren:
Xiaotian Chang, Yan Zhao, Xinfeng Yan, Jihong Pan, Kehua Fang, Lin Wang
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1186/​ar3429) contains supplementary material, which is available to authorized users.

Competing interests

The authors declare that they have no competing interests.

Authors' contributions

XC designed and executed the study and prepared the manuscript. JP and KF performed the genotyping. YZ and LW performed the Western blots and real time PCR. XY collected tissue samples. All authors have read and approved the final manuscript for publication.

Abstract

Introduction

Expression of TXNDC5, which is induced by hypoxia, stimulates cell proliferation and angiogenesis. Our previous study detected increased TXNDC5 expression in the synovial tissues of rheumatoid arthritis (RA) patients using proteomic methods. The current study investigated a pathogenic role for TXNDC5 in RA.

Method

Expression of TXNDC5 in synovial membranes was quantitatively analyzed by immunohistochemistry, Western blotting and real-time polymerase chain reaction (PCR). Serum TXNDC5 levels and serum anti-TXNDC5 antibody levels were determined using sandwich enzyme-linked immunosorbent assay (ELISA). A total of 96 single nucleotide polymorphisms (SNPs) in or near the TXNDC5 gene were genotyped using custom-designed Illumina 96-SNP VeraCode microassay. Allele frequencies and genotype frequencies of SNPs were assessed using a case-control design in a cohort of 267 Chinese patients with RA, 51 patients with ankylosing spondylitis (AS) and 160 healthy controls. Additional genotyping of 951 patients with RA and 898 healthy controls was performed for four SNPs (rs2277105, rs369086, rs443861 and rs11962800) using the TaqMan method.

Results

Real-time PCR, Western blotting and immunohistochemistry detected significantly higher TXNDC5 expression in the synovial tissues of RA patients compared to samples from patients with osteoarthritis (OA) or AS. ELISA detected significantly higher levels of TXNDC5 in the blood of RA patients compared to OA, AS and systemic lupus erythematosus patients, and healthy controls. ELISA did not detect significantly different levels of anti-TXNDC5 antibody in the blood of RA, OA and AS patients and healthy controls. A total of 9 SNPs (rs9505298, rs41302895, rs1225936, rs1225938, rs372578, rs443861, rs408014, rs9392189 and rs2743992) showed significant association with RA, while 16 SNPs (rs1044104, rs1225937, rs1225938, rs372578, rs89715, rs378963, rs1225944, rs1225947, rs1238994, rs369086, rs408014, rs368074, rs1225954, rs1225955, rs13209404 and rs3812162) showed significant association with AS. Taqman SNP assay demonstrated that rs443861 has an association with RA, which correlates with the microassay results.

Conclusions

TXNDC5 is up-regulated in synovial tissues of RA patients. TXNDC5 has a genetic effect on the risk of RA and AS.
Zusatzmaterial
Additional file 1: Supplementary materials and methods. This table summarizes the clinical data of patients with RA, OA and AS. (DOC 86 KB)
13075_2011_3199_MOESM1_ESM.DOC
Additional file 2: Supplementary results. This table provides the raw microarray data to perform association, LD and haplotype analysis. We genotyped 96 SNPs across the TXNDC5 gene from 267 Han Chinese patients with RA, 51 patients and 160 control individuals. All SNPs yielded genotype data, and the study sample success rate was 99.1%. (XLS 678 KB)
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Authors’ original file for figure 6
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