Suppression of urokinase receptor expression by bikunin is associated with inhibition of upstream targets of extracellular signal-regulated kinase-dependent cascade

Hiroshi Kobayashi, Mika Suzuki, Naohiro Kanayama, Takashi Nishida, Masaharu Takigawa, Toshihiko Terao

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Our laboratory showed that bikunin, a Kunitz-type protease inhibitor, suppresses 4β-phorbol 12-myristate 13-acetate (PMA)- or tumor necrosis factor-alpha (TNFα)-induced urokinase-type plasminogen activator (uPA) expression in different cell types. In addition to its effects on protease inhibition, bikunin could be modulating other cellular events associated with the metastatic cascade. To test this hypothesis, we examined whether bikunin was able to suppress the expression of uPA receptor (uPAR) mRNA and protein in a human chondrosarcoma cell line, HCS-2/8, and two human ovarian cancer cell lines, HOC-I and HRA. The present study showed that (a) bikunin suppresses the expression of constitutive and PMA-induced uPAR mRNA and protein in a variety of cell types; (b) an extracellular signal-regulated kinase (ERK) activation system is necessary for the PMA-induced increase in uPAR expression, as PD098059 and U0126, which prevent the activation of MEK1, reduce the uPAR expression; (c) bikunin markedly suppresses PMA-induced phosphorylation of ERK1/2 at the concentration that prevents uPAR expression, but does not reduce total ERK1/2 antigen level; (d) bikunin has no ability to inhibit overexpression of uPAR in cells treated with sodium vanadate; and (e) we further studied the inhibition of uPAR expression by stable transfection of HRA cells with bikunin gene, demonstrating that bikunin secretion is necessary for inhibition of uPAR expression. We conclude that bikunin downregulates constitutive and PMA-stimulated uPAR mRNA and protein possibly through suppression of upstream targets of the ERK-dependent cascade, independent of whether cells were treated with exogenous bikunin or transfected with bikunin gene.

Original languageEnglish
Pages (from-to)3945-3957
Number of pages13
JournalEuropean Journal of Biochemistry
Volume269
Issue number16
DOIs
Publication statusPublished - 2002

Fingerprint

Extracellular Signal-Regulated MAP Kinases
Urokinase-Type Plasminogen Activator
Acetates
Messenger RNA
Genes
Chemical activation
Cells
Urokinase Plasminogen Activator Receptors
Cell Line
Chondrosarcoma
Phosphorylation
Proteins
Vanadates
Protease Inhibitors
Ovarian Neoplasms
Transfection
Peptide Hydrolases
Down-Regulation
Tumor Necrosis Factor-alpha
phorbol-12-myristate

Keywords

  • Bikunin
  • Extracellular signal-regulated kinase
  • Tumor invasion
  • uPA receptor
  • Urokinase-type plasminogen activator

ASJC Scopus subject areas

  • Biochemistry

Cite this

Suppression of urokinase receptor expression by bikunin is associated with inhibition of upstream targets of extracellular signal-regulated kinase-dependent cascade. / Kobayashi, Hiroshi; Suzuki, Mika; Kanayama, Naohiro; Nishida, Takashi; Takigawa, Masaharu; Terao, Toshihiko.

In: European Journal of Biochemistry, Vol. 269, No. 16, 2002, p. 3945-3957.

Research output: Contribution to journalArticle

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AU - Suzuki, Mika

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AU - Nishida, Takashi

AU - Takigawa, Masaharu

AU - Terao, Toshihiko

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