A role for cyclic-GMP dependent protein kinase in anoikis

Yali Hou, Elsie Wong, Jessica Martin, Patricia V Schoenlein, Wolfgang R. Dostmann, Darren D Browning

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Anoikis is an essential process in which a loss of adhesion to the substratum alters intracellular signaling pathways that lead to apoptosis. Using phosphorylation of vasodilator stimulated phosphoprotein (VASP) as an indicator of cGMP-dependent protein kinase (PKG) activity in vivo, it was found that suspension of the colon epithelial cell line (CCD841) leads to rapid and transient activation of PKG that lasted several hours. The colon carcinoma lines SW480 and SW620 do not express endogenous PKG, but exogenously expressed PKG was similarly activated upon cell suspension. To determine whether PKG has a role in apoptosis following cell suspension, poly-ADP ribose polymerase (PARP) cleavage and propidium iodide staining were measured. After 24 h in suspension it was found that approximately 50% of CCD841 cells exhibited apoptosis, whereas apoptosis was not detected in either of the colon carcinoma cell lines. Inhibition of type 1 PKG by expression of a dominant negative PKG construct (G1αR-GFP), or by incubation with the PKG inhibitor peptide DT-2, blocked apoptosis in suspended CCD841 cells by approximately 50%. Furthermore, expression of exogenous PKG in SW620 and SW480 cells conferred partial sensitivity anoikis. Taken together these findings indicate that PKG has an important role in the induction of apoptosis following suspension of normal colon epithelial cells, and loss of PKG expression in colon tumor cells may contribute to resistance to anoikis.

Original languageEnglish (US)
Pages (from-to)882-888
Number of pages7
JournalCellular Signalling
Volume18
Issue number6
DOIs
StatePublished - Jun 1 2006

Fingerprint

Anoikis
Cyclic GMP-Dependent Protein Kinases
Suspensions
Apoptosis
Colon
Epithelial Cells
Carcinoma
Cell Line
Poly(ADP-ribose) Polymerases
Propidium
Phosphorylation
Staining and Labeling
Peptides

Keywords

  • Anoikis
  • Apoptosis
  • Cancer
  • Kinase
  • cGMP

ASJC Scopus subject areas

  • Cell Biology

Cite this

A role for cyclic-GMP dependent protein kinase in anoikis. / Hou, Yali; Wong, Elsie; Martin, Jessica; Schoenlein, Patricia V; Dostmann, Wolfgang R.; Browning, Darren D.

In: Cellular Signalling, Vol. 18, No. 6, 01.06.2006, p. 882-888.

Research output: Contribution to journalArticle

Hou, Yali ; Wong, Elsie ; Martin, Jessica ; Schoenlein, Patricia V ; Dostmann, Wolfgang R. ; Browning, Darren D. / A role for cyclic-GMP dependent protein kinase in anoikis. In: Cellular Signalling. 2006 ; Vol. 18, No. 6. pp. 882-888.
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