Signaling mechanisms responsible for lysophosphatidic acid-induced urokinase plasminogen activator expression in ovarian cancer cells

Hongbin Li, Xiaoqin Ye, Chitladda Mahanivong, Dafang Bian, Jerold Chun, Shuang Huang

Research output: Contribution to journalArticle

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Abstract

Lysophosphatidic acid (LPA) enhances urokinase plasminogen activator (uPA) expression in ovarian cancer cells; however, the molecular mechanisms responsible for this event have not been investigated. In this study, we used the invasive ovarian cancer SK-OV-3 cell line to explore the signaling molecules and pathways essential for LPA-induced uPA up-regulation. With the aid of specific inhibitors and dominant negative forms of signaling molecules, we determined that the Gi-associated pathway mediates this LPA-induced event. Moreover, constitutively active H-Ras and Raf-1-activating H-Ras mutant enhance uPA expression, whereas dominant negative H-Ras and Raf-1 block LPA-induced uPA up-regulation, suggesting that the Ras-Raf pathway works downstream of Gi to mediate this LPA-induced process. Surprisingly, dominant negative MEK1 or Erk2 displays only marginal inhibitory effect on LPA-induced uPA up-regulation, suggesting that a signaling pathway distinct from Raf-MEK1/2-Erk is the prominent pathway responsible for this process. In this report, we demonstrate that LPA activates NF-κB in a Ras-Raf-dependent manner and that blocking NF-κB activation with either non-phosphorylable IκB or dominant negative IκB kinase abolished LPA-induced uPA up-regulation and uPA promoter activation. Furthermore, introducing mutations to knock out the NF-κB binding site of the uPA promoter results in over 80% reduction in LPA-induced uPA promoter activation, whereas this activity is largely intact with the promoter containing mutations in the AP1 binding sites. Thus these results suggest that the Gi-Ras-Raf-NF-κB signaling cascade is responsible for LPA-induced uPA up-regulation in ovarian cancer cells.

Original languageEnglish (US)
Pages (from-to)10564-10571
Number of pages8
JournalJournal of Biological Chemistry
Volume280
Issue number11
DOIs
StatePublished - Mar 18 2005

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Plasminogen Activators
Urokinase-Type Plasminogen Activator
Ovarian Neoplasms
Cells
Up-Regulation
Chemical activation
Binding Sites
lysophosphatidic acid
Mutation
Molecules
Phosphotransferases
Cell Line

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Signaling mechanisms responsible for lysophosphatidic acid-induced urokinase plasminogen activator expression in ovarian cancer cells. / Li, Hongbin; Ye, Xiaoqin; Mahanivong, Chitladda; Bian, Dafang; Chun, Jerold; Huang, Shuang.

In: Journal of Biological Chemistry, Vol. 280, No. 11, 18.03.2005, p. 10564-10571.

Research output: Contribution to journalArticle

Li, Hongbin ; Ye, Xiaoqin ; Mahanivong, Chitladda ; Bian, Dafang ; Chun, Jerold ; Huang, Shuang. / Signaling mechanisms responsible for lysophosphatidic acid-induced urokinase plasminogen activator expression in ovarian cancer cells. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 11. pp. 10564-10571.
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abstract = "Lysophosphatidic acid (LPA) enhances urokinase plasminogen activator (uPA) expression in ovarian cancer cells; however, the molecular mechanisms responsible for this event have not been investigated. In this study, we used the invasive ovarian cancer SK-OV-3 cell line to explore the signaling molecules and pathways essential for LPA-induced uPA up-regulation. With the aid of specific inhibitors and dominant negative forms of signaling molecules, we determined that the Gi-associated pathway mediates this LPA-induced event. Moreover, constitutively active H-Ras and Raf-1-activating H-Ras mutant enhance uPA expression, whereas dominant negative H-Ras and Raf-1 block LPA-induced uPA up-regulation, suggesting that the Ras-Raf pathway works downstream of Gi to mediate this LPA-induced process. Surprisingly, dominant negative MEK1 or Erk2 displays only marginal inhibitory effect on LPA-induced uPA up-regulation, suggesting that a signaling pathway distinct from Raf-MEK1/2-Erk is the prominent pathway responsible for this process. In this report, we demonstrate that LPA activates NF-κB in a Ras-Raf-dependent manner and that blocking NF-κB activation with either non-phosphorylable IκB or dominant negative IκB kinase abolished LPA-induced uPA up-regulation and uPA promoter activation. Furthermore, introducing mutations to knock out the NF-κB binding site of the uPA promoter results in over 80{\%} reduction in LPA-induced uPA promoter activation, whereas this activity is largely intact with the promoter containing mutations in the AP1 binding sites. Thus these results suggest that the Gi-Ras-Raf-NF-κB signaling cascade is responsible for LPA-induced uPA up-regulation in ovarian cancer cells.",
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