Neumann, E. and Kullmann, F. and Judex, M. and Justen, H. P. and Wessinghage, D. and Gay, S. and Schoelmerich, Juergen and Mueller-Ladner, Ulf (2002) Identification of differentially expressed genes in rheumatoid arthritis by a combination of complementary DNA array and RNA arbitrarily primed-polymerase chain reaction. ARTHRITIS AND RHEUMATISM, 46 (1). pp. 52-63. ISSN 0004-3591
Full text not available from this repository. (Request a copy)Abstract
Objective. There is increasing evidence that T cell-independent pathways, such as the up-regulation of protooncogenes and the production of growth factors and matrix-degrading enzymes, lead to progressive destruction of affected joints. Therefore, identification of differentially regulated genes restricted to rheumatoid arthritis (RA) synovial fibroblasts is essential. A combination of RNA arbitrarily primed-polymerase chain reaction (RAP-PCR) and complementary DNA (cDNA) array with defined genes was used for a highly, sensitive differential screening using small amounts of RNA. Methods. RNA was extracted from cultured synovial fibroblasts obtained from 6 patients with RA and 6 patients with osteoarthritis (OA). RAP-PCR was performed using different arbitrary primers for first- and second-strand synthesis. PCRs were hybridized to cDNA array membranes. RA samples were compared with OA samples for differentially expressed genes. Results. In contrast to standard cDNA array, the identification of 12 differentially expressed genes in RA compared with OA (similar to6%) was possible. Differentially expressed genes of interest were confirmed using semiquantitative RT-PCR and in situ hybridization. Conclusion. Numerous variants of the differential display method and continuous improvements, including RAP-PCR, have proven to be both efficient and reliable for examining differentially regulated genes. Our results show that RAP-PCR combined with cDNA arrays is a suitable method for identifying differentially expressed genes in rheumatoid synovial fibroblasts, using very, small amounts of RNA.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | GROWTH-FACTOR-I; TUMOR-SUPPRESSOR GENE; HUMAN BREAST-CANCER; MESSENGER-RNAS; CELL-ADHESION; SYNOVIAL FIBROBLASTS; SIGNAL-TRANSDUCTION; FUNCTIONAL-ANALYSIS; EPITHELIAL-CELLS; CDNA ARRAYS; |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Innere Medizin I |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 15 Nov 2021 11:39 |
| Last Modified: | 15 Nov 2021 11:39 |
| URI: | https://pred.uni-regensburg.de/id/eprint/40694 |
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