Novel anti-inflammatory function of NSC95397 by the suppression of multiple kinases.
NSC95397 (2,3-bis-[(2-hydroxyethyl)thio]-1,4-naphthoquinone) is a CDC25 inhibitor with anti-cancer properties. Since the anti-inflammatory activity of this compound has not yet been explored, the aim of this study was to examine whether this compound is able to modulate the inflammatory process. Toll like receptor (TLR)-mediated inflammatory responses were induced by lipopolysaccharide (LPS), a TLR4 ligand, and pam3CSK, a TLR2 ligand, in peritoneal macrophages and RAW264.7. The molecular mechanism of NSC95397's anti-inflammatory activity was studied using immunoblotting analysis, nuclear fractionation, immunoprecipitation, overexpression strategies, luciferase reporter gene assays, and kinase assays. NSC95397 dose-dependently suppressed the production of nitric oxide (NO), tumor necrosis factor (TNF)-α, and prostaglandin (PG)E2, and diminished the mRNA expression of inflammatory genes such as inducible NO synthase (iNOS), cyclooxygenase (COX)-2, interferon (IFN)-β, and TNF-α in peritoneal macrophages and RAW264.7 cells that were stimulated by LPS and pam3CSK. This compound also clearly blocked the activation of NF-κB (p65), AP-1 (c-Fos/c-Jun), and IRF-3 in LPS-treated RAW264.7 cells and TRIF- and MyD88-overexpressing HEK293 cells. In addition, biochemical and molecular approaches revealed that this compound targeted AKT, IKKα/β, MKK7, and TBK1. Therefore, these results suggest that the anti-inflammatory function of NSC95397 can be attributed to its inhibition of multiple targets such as AKT, IKKα/β, MKK7, and TBK1.
Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
Yanyan
Y
Yang
Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
Woo Seok
WS
Yang
Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
Tao
T
Yu
Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
Young-Su
YS
Yi
Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
Jae Gwang
JG
Park
Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
Deok
D
Jeong
Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
Ji Hye
JH
Kim
Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
Jeong Su
JS
Oh
Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
Keejung
K
Yoon
Department of Veterinary Physiology, College of Veterinary Medicine, Biosafety Research Institute, Chonbuk National University, Jeonju 561-756, Republic of Korea. Electronic address: jhkim1@chonbuk.ac.kr.
Jong-Hoon
JH
Kim
Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea. Electronic address: jaecho@skku.edu.
Jae Youl
JY
Cho
D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009285:Naphthoquinones / Q000494:pharmacology*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D020687:cdc25 Phosphatases / Q000037:antagonists & inhibitors* / Q000378:metabolism
C451261:2,3-bis(2-hydroxyethylsulfanyl)-(1,4)naphthoquinone; D000894:Anti-Inflammatory Agents, Non-Steroidal; D009285:Naphthoquinones; D017346:Protein Serine-Threonine Kinases; D020687:cdc25 Phosphatases
AKT; Anti-inflammatory effects; IKKα/β; Inflammatory mediators; Multiple kinases; NSC95397