[논문]Redox Factor-1 Inhibits Cyclooxygenase-2 Expression via Inhibiting of p38 MAPK in the A549 Cells

Yoo, Dae-Goon; Kim, Cuk-Seong; Lee, Sang-Ki; Kim, Hyo-Shin; Cho, Eun-Jung; Park, Myoung-Soo; Lee, Sang-Do; Park, Jin-Bong; Jeon, Byeong-Hwa

doi:10.4196/kjpp.2010.14.3.139

Redox Factor-1 Inhibits Cyclooxygenase-2 Expression via Inhibiting of p38 MAPK in the A549 Cells 원문보기

The Korean journal of physiology & pharmacology : official journal of the Korean Physiological Society and the Korean Society of Pharmacology, v.14 no.3, 2010년, pp.139 - 144

Yoo, Dae-Goon (Department of Physiology, Infection Signaling Network Research Center, and Research Institute for Medical Sciences, School of Medicine, Chungnam National University) , Kim, Cuk-Seong (Cardiovascular Institute, University of Pittsburgh Medical Center) , Lee, Sang-Ki (Department of Sports Science, Chungnam National University) , Kim, Hyo-Shin (Department of Physiology, Infection Signaling Network Research Center, and Research Institute for Medical Sciences, School of Medicine, Chungnam National University) , Cho, Eun-Jung (Department of Physiology, Infection Signaling Network Research Center, and Research Institute for Medical Sciences, School of Medicine, Chungnam National University) , Park, Myoung-Soo (Department of Physiology, Infection Signaling Network Research Center, and Research Institute for Medical Sciences, School of Medicine, Chungnam National University) , Lee, Sang-Do (Department of Physiology, Infection Signaling Network Research Center, and Research Institute for Medical Sciences, School of Medicine, Chungnam National University) , Park, Jin-Bong (Department of Physiology, Infection) , Jeon, Byeong-Hwa

Abstract ▼ AI-Helper

In this study, we evaluated the role of apurinic/apyrimidinic endonuclease1/redox factor-1 (Ref-1) on the tumor necrosis factor-$\alpha$ (TNF-$\alpha$) induced cyclooxygenase-2 (COX-2) expression using A549 lung adenocarcinoma cells. TNF-$\alpha$ induced the expression of COX-2 in A549 cells, but did not induce BEAS-2B expression. The expression of COX-2 in A549 cells was TNF-$\alpha$ dose-dependent (5~100 ng/ml). TNF-$\alpha$-stimulated A549 cells evidenced increased Ref-1 expression in a dose-dependent manner. The adenoviral transfection of cells with AdRef-1 inhibited TNF-$\alpha$-induced COX-2 expression relative to that seen in the control cells ($Ad{\beta}gal$). Pretreatment with $10\;{\mu}M$ of SB203580 suppressed TNF-$\alpha$-induced COX-2 expression, thereby suggesting that p38 MAPK might be involved in COX-2 expression in A549 cells. The phosphorylation of p38 MAPK was increased significantly after 5 minutes of treatment with TNF-$\alpha$, reaching a maximum level at 10 min which persisted for up to 60 min. However, p38MAPK phosphorylation was markedly suppressed in the Ref-1-overexpressed A549 cells. Taken together, our results appear to indicate that Ref-1 negatively regulates COX-2 expression in response to cytokine stimulation via the inhibition of p38 MAPK phosphorylation. In the lung cancer cell lines, Ref-1 may be involved as an important negative regulator of inflammatory gene expression.

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문제 정의

The principal objective of this study was to elucidate the mechanisms responsible for the overexpression of Ref-1 and COX-2 in non-small cell lung cancer. These studies demonstrated the novel inhibitory role of Ref-1 on tumor COX-2 expression in response to TNF -a treatment in A549 lung adenocarcinoma cells.

제안 방법

As previously established Ref-1 inhibited COX-2 expression in A549 cells, we attempted to characterize the Ref-1-induced COX-2 expression in A549 cells. First, to evaluate the role of p38 MAPK on the expression of COX-2, the effects of SB203580, an inhibitor of p38 MAPK, were evaluated on the TNF-a-induced COX-2 expression. In this experiment, SB203580 was pretreated for 30 min prior to TNF-a treatment, and the cells were exposed to TNF -a for 6 hours.
determined. In this study, we evaluated the role of Ref-1 on TNF-a induced COX-2 expression using the A549 lung adenocarcinoma cells.
in non-small cell lung cancer. These studies demonstrated the novel inhibitory role of Ref-1 on tumor COX-2 expression in response to TNF -a treatment in A549 lung adenocarcinoma cells.

대상 데이터

from Sigma. SB203580, a specific inhibitor of p38 MAPK, was purchased from Tocris Bioscience (Bristol, UK). All reagents were obtained from Sigma-Aldrich unless stated otherwise.

데이터처리

Values are expressed as the means±S.E.M. Statistical evaluations were conducted using the Student's t-test, with pv0.05 being considered significant.

성능/효과

This finding is consistent with a previous report showing increased COX-2 expression in transformed cells [24]. In the preliminary study we also confirmed that NCI-H₄₆0 and VMRC-LCD human lung carcinoma cell lines did not evidence COX-2 expression in response to TNF-a (Data not shown). Therefore, we selected A549 cell lines in this experiment.

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