LYG-202 augments tumor necrosis factor-α-induced apoptosis via attenuating casein kinase 2-dependent nuclear factor-κB pathway in HepG2 cells

Mol Pharmacol. 2012 Nov;82(5):958-71. doi: 10.1124/mol.112.079848. Epub 2012 Aug 21.

Abstract

Tumor necrosis factor-α (TNF-α) is being used as an antineoplastic agent in treatment regimens of patients with locally advanced solid tumors, but TNF-α alone is only marginally active. In clinical use, it is usually combined with other chemical agents to increase its tumor response rate. Our previous studies reported that LYG-202 (5-hydroxy-8-methoxy-7-(4-(4-methylpiperazin-1-yl)butoxy)-2-phenyl-4H-chromen-4-one), a synthesized flavonoid with a piperazine substitution, has antiproliferative, antiangiogenic, and proapoptotic activities in multiple cancer cell lines. Here we evaluated the antineoplastic effect of TNF-α and analyzed the mechanism underlying its combination with LYG-202. Our results indicated that LYG-202 significantly increased the cytostatic and proapoptotic activity of TNF-α in HepG2 cells and heightened the protein level of apoptosis-related genes including caspase-3, caspase-8/9, cleaved poly(ADP-ribose) polymerase, and Bid. The fact that LYG-202 had a fitness score similar to that of the casein kinase 2 (CK2) inhibitor naphthyridine-8-carboxylate (CX-4945) implied to us that it may serve as a potential candidate for CK2 inhibitor, and the kinase activity assay suggested that LYG-202 significantly inhibited CK2 activity. Moreover, the electrophoretic mobility shift assay and luciferase assay showed that LYG-202 blocked the TNF-α-induced nuclear factor-κB (NF-κB) survival signaling pathway primarily by inactivating protein kinase CK2. In murine xenograft models, we also found that LYG-202 enhanced TNF-α antineoplastic activity and inhibited CK2 activity and NF-κB-regulated antiapoptotic gene expression. All these results showed that LYG-202 enhanced TNF-α-induced apoptosis by attenuating the CK2-dependent NF-κB pathway and probably is a promising agent in combination with TNF-α.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Active Transport, Cell Nucleus
  • Animals
  • Antineoplastic Agents / pharmacology*
  • Apoptosis / drug effects*
  • Casein Kinase II / physiology*
  • Caspases / metabolism
  • Cell Nucleus / metabolism
  • Cell Proliferation / drug effects
  • Drug Synergism
  • Electrophoretic Mobility Shift Assay
  • Flavones / pharmacology*
  • Genes, Reporter
  • Hep G2 Cells
  • Humans
  • I-kappa B Kinase / antagonists & inhibitors
  • I-kappa B Proteins / metabolism
  • Luciferases / genetics
  • Male
  • Mice
  • Molecular Docking Simulation
  • NF-KappaB Inhibitor alpha
  • NF-kappa B / genetics
  • NF-kappa B / physiology*
  • Phosphorylation
  • Piperazines / pharmacology*
  • Signal Transduction
  • Transcription Factor RelA / metabolism
  • Transcription, Genetic / drug effects
  • Tumor Necrosis Factor-alpha / pharmacology*
  • Xenograft Model Antitumor Assays

Substances

  • Antineoplastic Agents
  • Flavones
  • I-kappa B Proteins
  • LYG 202
  • NF-kappa B
  • NFKBIA protein, human
  • Nfkbia protein, mouse
  • Piperazines
  • Transcription Factor RelA
  • Tumor Necrosis Factor-alpha
  • NF-KappaB Inhibitor alpha
  • Luciferases
  • Casein Kinase II
  • I-kappa B Kinase
  • Caspases