title: 12-O-Tetradecanoylphorbol-13-Acetate Induces Keratin 8 Phosphorylation and Reorganization via Expression of Transglutaminase-2.
abstract: The stiffness of cancer cells is attributable to intermediate filaments such as keratin. Perinuclear reorganization via phosphorylation of specific serine residue in keratin is implicated in the deformability of metastatic cancer cells including the human pancreatic carcinoma cell line (PANC-1). 12-O-Tetradecanoylphorbol-13-acetate (TPA) is a potent tumor promoter and protein kinase C (PKC) activator. However, its effects on phosphorylation and reorganization of keratin 8 (K8) are not well known. Therefore, we examined the underlying mechanism and effect of TPA on K8 phosphorylation and reorganization. TPA induced phosphorylation and reorganization of K8 and transglutaminase-2 (Tgase-2) expression in a time- and dose-dependent manner in PANC-1 cells. These effects peaked after 45 min and 100 nM of TPA treatment. We next investigated, using cystamine (CTM), Tgase inhibitor, and Tgase-2 gene silencing, Tgase-2's possible involvement in TPA-induced K8 phosphorylation and reorganization. We found that Tgase-2 gene silencing inhibited K8 phosphorylation and reorganization in PANC-1 cells. Tgase-2 gene silencing, we additionally discovered, suppressed TPA-induced migration of PANC-1 cells and Tgase-2 overexpression induced migration of PANC-1 cells. Overall, these results suggested that TPA induced K8 phosphorylation and reorganization via Tgase-2 expression in PANC-1 cells.
authors.affiliation: BK21PLUS R-FIND Team, College of Pharmacy, Dongguk University, Seoul 100-715, Korea.
authors.forename: Eun Ji
authors.initials: EJ
authors.lastname: Lee
authors.affiliation: BK21PLUS R-FIND Team, College of Pharmacy, Dongguk University, Seoul 100-715, Korea.
authors.forename: Mi Kyung
authors.initials: MK
authors.lastname: Park
authors.affiliation: BK21PLUS R-FIND Team, College of Pharmacy, Dongguk University, Seoul 100-715, Korea.
authors.forename: Hyun Ji
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: BK21PLUS R-FIND Team, College of Pharmacy, Dongguk University, Seoul 100-715, Korea.
authors.forename: June Hee
authors.initials: JH
authors.lastname: Kang
authors.affiliation: BK21PLUS R-FIND Team, College of Pharmacy, Dongguk University, Seoul 100-715, Korea.
authors.forename: You Ri
authors.initials: YR
authors.lastname: Kim
authors.affiliation: BK21PLUS R-FIND Team, College of Pharmacy, Dongguk University, Seoul 100-715, Korea.
authors.forename: Gyeoung Jin
authors.initials: GJ
authors.lastname: Kang
authors.affiliation: BK21PLUS R-FIND Team, College of Pharmacy, Dongguk University, Seoul 100-715, Korea.
authors.forename: Hyun Jung
authors.initials: HJ
authors.lastname: Byun
authors.affiliation: BK21PLUS R-FIND Team, College of Pharmacy, Dongguk University, Seoul 100-715, Korea.
authors.forename: Chang Hoon
authors.initials: CH
authors.lastname: Lee
keywords: 12-O-Tetradecanoylphorbol-13-acetate; Keratin 8 phosphorylation; PANC-1 cells; Perinuclear reorganization; Transglutaminase-2
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