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Longterm Culture of Telomerase-Transduced Rheumatoid Arthritis Fibroblast-like Synoviocytes Display a Distinct Gene Expression Pattern YUBO SUN, GARY FIRESTEIN, DAVID BOYLE, HELEN GRUBER, and HERMAN CHEUNG
ABSTRACT. Methods. RA 516 FLS were transduced by hTERT and the replicative properties of hTERT-RA 516 FLS were evaluated by repeated expansion. Gene expressions in hTERT-RA 516 FLS (passage 8) were compared with the gene expressions in hTERT-osteoarthritis (OA) 13A FLS (passage 8) by microarrays and RT-PCR. After continuous expansion in culture for an additional 4 months, gene expression was examined again using real-time RT-PCR and microarray. Results. While primary RA 516 FLS stopped dividing after repeated culture for about 120 days, hTERT-RA 516 FLS continued to grow at a steady rate. The hTERT-RA 156 FLS displayed a distinct gene expression pattern different from hTERT-OA 13A FLS. Several putative autoantigens and cytokine A2/monocyte chemoattractant protein-1 were expressed at significantly higher levels in longterm culture of hTERT-RA 516 FLS. Conclusion. Telomerase-transduced RA FLS offer an alternative cell model for the study of RA and for examination of cellular/genetic alterations in RA FLS. Our findings provide further support for the notion that RA FLS are altered cells. (First Release Sept 15 2007; J Rheumatol 2007;34:1959-70) Key Indexing Terms:
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From the Department of Orthopaedic Surgery, Carolinas Medical Center, Charlotte, North Carolina; the Division of Rheumatology, Allergy and Immunology, University of California San Diego, School of Medicine, La Jolla, California; and the Department of Biomedical Engineering, University of Miami, Coral Gables, Florida, Research Service and Geriatric Research, Education, and Clinical Center, Veterans Affairs Medical Center, Miami, Florida, USA. Y. Sun, PhD; H.E. Gruber, PhD, Department of Orthopaedic Surgery, Carolinas Medical Center; G.S. Firestein, MD; D.L. Boyle, PhD, Division of Rheumatology, Allergy and Immunology, University of California San Diego, School of Medicine; H.S. Cheung, PhD, Department of Biomedical Engineering, University of Miami, Coral Gables, Florida, Research Service and Geriatric Research, Education, and Clinical Center, Veterans Affairs Medical Center, Miami. Address reprint requests to Dr. Y. Sun, Department of Orthopaedic Surgery, Biology Division, Cannon Research 304, Carolinas Medical Center, Charlotte, NC 28232, USA. E-mail: yubo.sun@carolinashealthcare.org Accepted for publication June 29, 2007. |
