title: The role of PKC isoforms in the inhibition of NF-κB activation by vitamin K2 in human hepatocellular carcinoma cells.
abstract: Vitamin K (VK) has diverse protective effects against osteoporosis, atherosclerosis and carcinogenesis. We recently reported that menatetrenone, a VK2 analogue, suppressed nuclear factor (NF)-κB activation in human hepatoma cells. Although NF-κB is regulated by isoforms of protein kinase C (PKC), the involvement of PKCs in VK2-mediated NF-κB inhibition remains unknown. Therefore, the effects of VK2 on the activation and the kinase activity of each PKC isoform were investigated. The human hepatoma Huh7 cells were treated with PKC isoform-specific inhibitors and/or siRNAs against each PKC isoform with or without 12-O-tetradecanoylphorbol-13-acetate (TPA). VK2 inhibited the TPA-induced NF-κB activation in Huh7 cells. NF-κB activity was inhibited by the pan-PKC inhibitor Ro-31-8425, but not by the PKCα-specific inhibitor Gö6976. The knockdown of individual PKC isoforms including PKCα, δ and ɛ showed only marginal effects on the NF-κB activity. However, the knockdown of both PKCδ and PKCɛ, together with treatment with a PKCα-specific inhibitor, depressed the NF-κB activity. VK2 suppressed the PKCα kinase activity and the phosphorylation of PKCɛ after TPA treatment, but neither the activation nor the enzyme activity of PKCδ was affected. The knockdown of PKCɛ abolished the TPA-induced phosphorylation of PKD1, and the effects of PKD1 knockdown on NF-κB activation were similar to those of PKCɛ knockdown. Collectively, all of the PKCs, including α, δ and ɛ, and PKD1 are involved in the TPA-mediated activation of NF-κB. VK2 inhibited the NF-κB activation through the inhibition of PKCα and ɛ kinase activities, as well as subsequent inhibition of PKD1 activation.
authors.affiliation: Department of Internal Medicine, Saga Medical School, Saga University 5-1-1 Nabeshima, Saga 849-8501, Japan.
authors.forename: JingHe
authors.initials: J
authors.lastname: Xia
authors.forename: Sachiko
authors.initials: S
authors.lastname: Matsuhashi
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Hamajima
authors.forename: Shinji
authors.initials: S
authors.lastname: Iwane
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Takahashi
authors.forename: Yuichiro
authors.initials: Y
authors.lastname: Eguchi
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Mizuta
authors.forename: Kazuma
authors.initials: K
authors.lastname: Fujimoto
authors.forename: Shun'ichi
authors.initials: S
authors.lastname: Kuroda
authors.forename: Iwata
authors.initials: I
authors.lastname: Ozaki
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000378:metabolism; D045744:Cell Line, Tumor; D004791:Enzyme Inhibitors / Q000494:pharmacology; D055785:Gene Knockdown Techniques; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000378:metabolism; D008301:Maleimides / Q000494:pharmacology; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D051745:Protein Kinase C-delta / Q000235:genetics / Q000378:metabolism; D051744:Protein Kinase C-epsilon / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D024482:Vitamin K 2 / Q000494:pharmacology*
chemical_list: D004791:Enzyme Inhibitors; D007211:Indoles; D008301:Maleimides; D016328:NF-kappa B; D034741:RNA, Small Interfering; D024482:Vitamin K 2; C071924:Ro 31-8425; C089212:protein kinase D; D011493:Protein Kinase C; D051745:Protein Kinase C-delta; D051744:Protein Kinase C-epsilon; D013755:Tetradecanoylphorbol Acetate