Anti-inflammatory effect of pitavastatin on NF-kappaB activated by TNF-alpha in hepatocellular carcinoma cells.
As nuclear factor-kappa B (NF-kappaB) is essential for promoting inflammation-associated cancer, it is a potential target for cancer prevention in chronic inflammatory diseases. Here we examined the anti-inflammatory effect of pitavastatin, a 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, on NF-kappaB activated by TNF-alpha in hepatocellular carcinoma (HCC) cells. Western blot revealed that the treatment of Huh 7 cells with pitavastatin at 0.1 microM inhibited the nuclear expression of NF-kappaB p65 induced by TNF-alpha. Furthermore, electrophoretic mobility shift assay showed that after the cells were incubated with pitavastatin alone or with pitavastatin and TNF-alpha for 24 h, pitavastatin significantly decreased the DNA binding activity of NF-kappaB induced by TNF-alpha. Subsequently, luciferase assay revealed that pitavastatin suppressed the transcriptional activity of the NF-kappaB promoter, which was clearly related to the HMG-CoA reductase activity because the addition of mevalonic acid (MEV) elevated the TNF-alpha activity. Moreover, the Rho kinase inhibitor Y27632 had no major effect on the NF-kappaB inhibitory activity of pitavastatin. The inhibitory effect of pitavastatin is possibly independent of the Rho kinase pathway in inflammation-associated HCC cells is. Finally, the addition of TNF-alpha significantly increased IL-6 protein production, which was suppressed by the addition of pitavastatin. These results suggest that pitavastatin at a low dose (0.1 microM) inhibits NF-kappaB activation and decreases IL-6 production induced by TNF-alpha, and is therefore expected to be a new strategy for treating HCC.
Wang
Juyong
J
Department of Clinical Laboratory Medicine, Faculty of Medicine, Toyama University, Sugitani, Japan.
Tokoro
Takashi
T
Higa
Susumu
S
Kitajima
Isao
I
D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000235:genetics*; D045744:Cell Line, Tumor; D024202:Electrophoretic Mobility Shift Assay; D004797:Enzyme-Linked Immunosorbent Assay; D017930:Genes, Reporter / Q000235:genetics; D006801:Humans; D007202:Indicators and Reagents; D015850:Interleukin-6 / Q000096:biosynthesis; D047908:Intracellular Signaling Peptides and Proteins; D008113:Liver Neoplasms / Q000235:genetics*; D008156:Luciferases / Q000235:genetics; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000378:metabolism; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000235:genetics; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011804:Quinolines / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects; D013778:Tetrazolium Salts / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology*; D054460:rho-Associated Kinases
C476329:2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H tetrazolium monosodium salt; D000893:Anti-Inflammatory Agents; D007202:Indicators and Reagents; D015850:Interleukin-6; D047908:Intracellular Signaling Peptides and Proteins; D016328:NF-kappa B; D009687:Nuclear Proteins; D011804:Quinolines; D013778:Tetrazolium Salts; D014409:Tumor Necrosis Factor-alpha; D008156:Luciferases; D017346:Protein Serine-Threonine Kinases; D054460:rho-Associated Kinases; C108475:pitavastatin