TLR4-MyD88-TRAF6-TAK1 Complex-Mediated NF-κB Activation Contribute to the Anti-Inflammatory Effect of V8 in LPS-Induced Human Cervical Cancer SiHa Cells

Inflammation. 2016 Feb;39(1):172-181. doi: 10.1007/s10753-015-0236-8.

Abstract

The synthetic compound 7-4-[Bis-(2-hydroxyethyl)-amino]-butoxy-5-hydroxy-8-methoxy-2-phenylchromen-4-one (V8) is a novel flavonoid-derived compound. In this study, we investigated the effects of V8 on Toll-like receptor 4 (TLR4)-mediated inflammatory reaction in human cervical cancer SiHa cells and lipopolysaccharide (LPS)-induced TLR4 activity in cervical cancer SiHa (HPV16+) cells, but not in HeLa (HPV18+) and C33A (HPV-) cells. In addition, V8 inhibited LPS-induced expression of TLR4, MyD88, TRAF6 and phosphorylation of TAK1, and their interaction with TLR4 in SiHa cells, resulting in an inhibition of TLR4-MyD88-TRAF6-TAK1 complex. Moreover, V8 blocked LPS-induced phosphorylation of IκB and IKK, resulting in inhibition of the nuclear translocation of P65-NF-κB in SiHa cells. We also found that V8 reduced the expression of NF-κB target genes, such as those for COX-2, iNOS, IL-6, IL-8, CCL-2, and TNF-α in LPS-stimulated SiHa cells. These results suggested that V8 exerted an anti-inflammatory effect on SiHa cells by inhibiting the TLR4-MyD88-TRAF6-TAK1 complex-mediated NF-κB activation.

Keywords: HPV16; NF-κB; TLR4; V8; cervical cancer; inflammation.

MeSH terms

  • Active Transport, Cell Nucleus / drug effects
  • Anti-Inflammatory Agents / pharmacology*
  • Cell Line, Tumor
  • Female
  • Flavones / pharmacology*
  • Flavonoids / pharmacology*
  • HeLa Cells
  • Humans
  • I-kappa B Kinase / metabolism
  • Lipopolysaccharides / pharmacology*
  • MAP Kinase Kinase Kinases / antagonists & inhibitors*
  • MAP Kinase Kinase Kinases / metabolism
  • Myeloid Differentiation Factor 88 / antagonists & inhibitors*
  • Myeloid Differentiation Factor 88 / biosynthesis
  • Phosphorylation / drug effects
  • TNF Receptor-Associated Factor 6 / antagonists & inhibitors*
  • TNF Receptor-Associated Factor 6 / biosynthesis
  • Toll-Like Receptor 4 / antagonists & inhibitors*
  • Toll-Like Receptor 4 / biosynthesis
  • Transcription Factor RelA / metabolism
  • Uterine Cervical Neoplasms

Substances

  • 7-(4-(bis-(2-hydroxyethyl)amino)butoxy)-5-hydroxy-8-methoxy-2-phenylchromen-4-one
  • Anti-Inflammatory Agents
  • Flavones
  • Flavonoids
  • Lipopolysaccharides
  • MYD88 protein, human
  • Myeloid Differentiation Factor 88
  • RELA protein, human
  • TLR4 protein, human
  • TNF Receptor-Associated Factor 6
  • Toll-Like Receptor 4
  • Transcription Factor RelA
  • I-kappa B Kinase
  • MAP Kinase Kinase Kinases
  • MAP kinase kinase kinase 7