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Inhibition of NF-κB Renders Human Juvenile Costal Chondrocyte Cell Lines Sensitive to TNF-α-Mediated Cell Death

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Abstract

Background. Recently, therapeutics employing knowledge on various signaling pathways are being developed, with NF-κB being one of the most promising targets. NF-κB has been suggested to play a role not only in the induction of inflammatory mediators, but also in the protection from cell death. Objectives. This study pursued the role of the NF-κB pathway in the regulation of chondrocyte death induced by tumor necrosis factor alpha (TNF-α) and of the pertinent target molecules involved. Methods. The human chondrocyte cell line C28/I2 was used for the experiment. Chondrocytes were transduced with adenovirus-encoding IkappaB (IκB) superrepressor which inhibits NF-κB activation, and treated with TNF-α. The proportion of cell death was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazdium bromide (MTT) assay. Activation of p38 mitogen activated protein (MAP) kinase and phosphatidylinositol 3-kinase (PI3K) by TNF-α was inhibited with SB202190 and Ly 294002 respectively. The expression of apoptosis related protein was analyzed with western blot assay, and the activation of c-Jun N-terminal kinase (JNK) by solid-phase kinase assay. Results. Treatment with TNF-α led to cell death in 23% and 50% of ad-IκB-SR infected chondrocytes after 24 and 72 h respectively. The expression of Bcl-XL, Bcl-2, and XIAP significantly decreased, and activation of JNK was prolonged for up to 6 h in infected cells treated with TNF-α. Preincubation with p38 inhibitor or PI3K inhibitor before TNF-α led to a significant increase in cell death in ad-IκB-SR transduced chondrocytes, resulting in 53% and 30% cell death after 24 h for p38 inhibitor and PI3K inhibitor respectively. Conclusion. In our experimental system, specific inhibition of NF-κB activation rendered chondrocytes susceptible to cell death induced by TNF-α. The cell death was enhanced by inhibition of another signaling pathway such as p38 MAP kinase or PI3K. The expression of Bcl-XL, Bcl-2 and XIAP and activation of JNK were affected by ad-IκB-SR transduction, implying a role in the NF-κB regulated cell survival signaling in human chondrocytes.

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Acknowledgement

This study was supported by a grant from the Korea Health 21 R & D project, Ministry of Health and Welfare (grant number 01-PJ3-PG6-01GN11-0002).

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Authors and Affiliations

  1. Department of Internal Medicine, Hallym University Kangdong Sacred Heart Hospital, 445 Gil-dong, Kangdong-gu, Seoul, 134-701, South Korea

    Ho Sung Yoon

  2. Division of rheumatology, Department of Internal Medicine, Hallym University Sacred Heart Hospital, 896, Pyongchondong, Dongan-gu, Anyang, Kyunggi-do, 431-070, South Korea

    Hyun Ah Kim

  3. Department of Internal Medicine, Seoul National University College of medicine, 28, Yongon-dong, Chongno-gu, Seoul, 110-799, South Korea

    Yeong Wook Song

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  1. Ho Sung Yoon
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Correspondence to Hyun Ah Kim.

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Yoon, H.S., Kim, H.A. & Song, Y.W. Inhibition of NF-κB Renders Human Juvenile Costal Chondrocyte Cell Lines Sensitive to TNF-α-Mediated Cell Death. Rheumatol Int 26, 201–208 (2006). https://doi.org/10.1007/s00296-004-0562-x

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  • DOI: https://doi.org/10.1007/s00296-004-0562-x

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