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Protein kinase Cepsilon is important for migration of neuroblastoma cells

Stensman, Helena LU and Larsson, Christer LU (2008) In BMC Cancer 8.
Abstract
Background: Migration is important for the metastatic capacity and thus for the malignancy of cancer cells. There is limited knowledge on regulatory factors that promote the migration of neuroblastoma cells. This study investigates the hypothesis that protein kinase C (PKC) isoforms regulate neuroblastoma cell motility. Methods: PKC isoforms were downregulated with siRNA or modulated with activators and inhibitors. Migration was analyzed with scratch and transwell assays. Protein phosphorylation and expression levels were measured with Western blot. Results: Stimulation with 12-O-tetradecanoylphorbol-13-acetate (TPA) induced migration of SK-N-BE(2) C neuroblastoma cells. Treatment with the general protein kinase C (PKC) inhibitor GF109203X... (More)
Background: Migration is important for the metastatic capacity and thus for the malignancy of cancer cells. There is limited knowledge on regulatory factors that promote the migration of neuroblastoma cells. This study investigates the hypothesis that protein kinase C (PKC) isoforms regulate neuroblastoma cell motility. Methods: PKC isoforms were downregulated with siRNA or modulated with activators and inhibitors. Migration was analyzed with scratch and transwell assays. Protein phosphorylation and expression levels were measured with Western blot. Results: Stimulation with 12-O-tetradecanoylphorbol-13-acetate (TPA) induced migration of SK-N-BE(2) C neuroblastoma cells. Treatment with the general protein kinase C (PKC) inhibitor GF109203X and the inhibitor of classical isoforms Go6976 inhibited migration while an inhibitor of PKC beta isoforms did not have an effect. Downregulation of PKC epsilon, but not of PKC alpha or PKC delta, with siRNA led to a suppression of both basal and TPA-stimulated migration. Experiments using PD98059 and LY294002, inhibitors of the Erk and phosphatidylinositol 3-kinase (PI3K) pathways, respectively, showed that PI3K is not necessary for TPA-induced migration. The Erk pathway might be involved in TPA-induced migration but not in migration driven by PKC epsilon. TPA induced phosphorylation of the PKC substrate myristoylated alanine-rich C kinase substrate (MARCKS) which was suppressed by the PKC inhibitors. Treatment with siRNA oligonucleotides against different PKC isoforms before stimulation with TPA did not influence the phosphorylation of MARCKS. Conclusion: PKC epsilon is important for migration of SK-N-BE(2) C neuroblastoma cells. Neither the Erk pathway nor MARCKS are critical downstream targets of PKC epsilon but they may be involved in TPA-mediated migration. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
BMC Cancer
volume
8
publisher
BioMed Central
external identifiers
  • wos:000262704100002
  • scopus:58149373711
ISSN
1471-2407
DOI
10.1186/1471-2407-8-365
language
English
LU publication?
yes
id
954ed32d-a07e-406c-ba0b-caabf4a2fa77 (old id 1375983)
date added to LUP
2009-04-24 13:48:12
date last changed
2017-03-12 03:41:57
@article{954ed32d-a07e-406c-ba0b-caabf4a2fa77,
  abstract     = {Background: Migration is important for the metastatic capacity and thus for the malignancy of cancer cells. There is limited knowledge on regulatory factors that promote the migration of neuroblastoma cells. This study investigates the hypothesis that protein kinase C (PKC) isoforms regulate neuroblastoma cell motility. Methods: PKC isoforms were downregulated with siRNA or modulated with activators and inhibitors. Migration was analyzed with scratch and transwell assays. Protein phosphorylation and expression levels were measured with Western blot. Results: Stimulation with 12-O-tetradecanoylphorbol-13-acetate (TPA) induced migration of SK-N-BE(2) C neuroblastoma cells. Treatment with the general protein kinase C (PKC) inhibitor GF109203X and the inhibitor of classical isoforms Go6976 inhibited migration while an inhibitor of PKC beta isoforms did not have an effect. Downregulation of PKC epsilon, but not of PKC alpha or PKC delta, with siRNA led to a suppression of both basal and TPA-stimulated migration. Experiments using PD98059 and LY294002, inhibitors of the Erk and phosphatidylinositol 3-kinase (PI3K) pathways, respectively, showed that PI3K is not necessary for TPA-induced migration. The Erk pathway might be involved in TPA-induced migration but not in migration driven by PKC epsilon. TPA induced phosphorylation of the PKC substrate myristoylated alanine-rich C kinase substrate (MARCKS) which was suppressed by the PKC inhibitors. Treatment with siRNA oligonucleotides against different PKC isoforms before stimulation with TPA did not influence the phosphorylation of MARCKS. Conclusion: PKC epsilon is important for migration of SK-N-BE(2) C neuroblastoma cells. Neither the Erk pathway nor MARCKS are critical downstream targets of PKC epsilon but they may be involved in TPA-mediated migration.},
  author       = {Stensman, Helena and Larsson, Christer},
  issn         = {1471-2407},
  language     = {eng},
  publisher    = {BioMed Central},
  series       = {BMC Cancer},
  title        = {Protein kinase Cepsilon is important for migration of neuroblastoma cells},
  url          = {http://dx.doi.org/10.1186/1471-2407-8-365},
  volume       = {8},
  year         = {2008},
}