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Dual knockdown of p65 and p50 subunits of NF-kappaB by siRNA inhibits the induction of inflammatory cytokines and significantly enhance apoptosis in human primary synoviocytes treated with tumor necrosis factor-alpha

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Abstract

In order to develop an anti-NF-κB siRNA as a novel class of anti-inflammatory drug, we have isolated a highly efficient siRNA targeting the p65 (RelA) subunit of NF-κB, hereafter named REL1096. To determine whether down-regulation of p65 by REL1096 can block the induction of inflammatory cytokines after treatment with tumor necrosis factor-alpha (TNF-α), human primary fibroblast-like synoviocytes were isolated from patients with rheumatoid arthritis. When treated with REL1096, the TNF-mediated induction of downstream target genes such as inflammatory cytokines, chemokines, and anti-apoptosis genes was drastically inhibited. To enhance the inhibitory effect of REL1096, cells were treated with siRNA targeting the p50 subunit of NF-κB together with REL1096. In addition to effective downregulation of inflammatory cytokines, knockdown of both p65 and p50 resulted in much more extensive apoptosis when compared to cells treated with either REL1096 or p50-siRNA alone. Thus, our results provide evidence for the potential use of siRNA targeting NF-κB as an effective means to treat rheumatoid arthritis. In addition to effective amelioration of synovial inflammation by downregulation of inflammatory cytokines, increased apoptosis by dual knockdown of p65 and p50 may prove advantageous in preventing invasiveness and destructiveness of hyperplastic synoviocytes.

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Abbreviations

TNF-α:

Tumor necrosis factor-alpha

RA:

Rheumatoid arthritis

IκB:

Inhibitor κB

IKK:

IκB kinase complex

CIA:

Collagen-induced arthritis

FLSs:

Fibroblast-like synoviocytes

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Acknowledgements

This work was supported by the International Joint R&D Program (10014823) from the Ministry of Commerce, Industry, and Energy in Korea to Y-C Choi.

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Authors and Affiliations

  1. Gene2Drug Research Center, Bioneer Corporation, 49-3, Munpyeong-dong, Daedeok-gu, Daejeon, 306-220, Republic of Korea

    So-Rim Choung, Eun-Ju Lee, Mi Young Lee & Young-Chul Choi

  2. National Genome Information Center, KRIBB, 52 Eoeun-dong, Yuseong-gu, Daejeon, 305-333, Republic of Korea

    Young Joo Kim

  3. Daejeon St. Mary’s Hospital, College of Medicine, Catholic University of Korea, Seoul, 301-723, Republic of Korea

    Ui Jin Lee, K. V. Bhanu Prakash & Chang-Whan Han

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  1. Ui Jin Lee
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  2. So-Rim Choung
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  3. K. V. Bhanu Prakash
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  4. Eun-Ju Lee
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  5. Mi Young Lee
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  6. Young Joo Kim
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  7. Chang-Whan Han
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  8. Young-Chul Choi
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Corresponding authors

Correspondence to Chang-Whan Han or Young-Chul Choi.

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Ui Jin Lee and So-Rim Choung contributed equally to this work.

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Lee, U.J., Choung, SR., Prakash, K.V.B. et al. Dual knockdown of p65 and p50 subunits of NF-kappaB by siRNA inhibits the induction of inflammatory cytokines and significantly enhance apoptosis in human primary synoviocytes treated with tumor necrosis factor-alpha. Mol Biol Rep 35, 291–298 (2008). https://doi.org/10.1007/s11033-007-9084-4

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  • DOI: https://doi.org/10.1007/s11033-007-9084-4

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