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Breast Cancer Research and Treatment

, Volume 91, Issue 1, pp 35–45 | Cite as

Inositol hexaphosphate (IP6) blocks proliferation of human breast cancer cells through a PKCδ-dependent increase in p27Kip1 and decrease in retinoblastoma protein (pRb) phosphorylation

  • Ivana Vucenik
  • Gayatri Ramakrishna
  • Kwanchanit Tantivejkul
  • Lucy M. Anderson
  • Danica Ramljak
Report

Summary

Inositol hexaphosphate (IP6) is a naturally occurring polyphosphorylated carbohydrate with demonstrated anti-proliferative and anti-cancer activity in mammary cells. We hypothesized that IP6 modulates cell cycle proteins by action on cytoplasmic signaling molecules. The effects of both pharmacological (2 mM) and physiological (100 μM) doses of IP6 on major PKC isoforms (PKCα, δ, ε, β and ζ), PI3-K/Akt and ras/Erk1/2 were evaluated. Treatment of MCF-7 human breast cancer cells with 2 mM IP6 for 24 h caused a 3.1-fold increase in the expression of anti-proliferative PKCδ. Similar results were observed with 100 μM IP6 at only 30–60 min post-treatment. IP6 also caused an increase in PKCδ activity, shown by its translocation from cytosol to membrane. No changes in expression of PKC α, δ, ε, β and ζ were detected. Additionally, IP6 caused a decrease of Erk1/2 and Akt activity. Among cell cycle control proteins, IP6 resulted in increased p27Kip1 protein levels and marked reduction of pRb phosphorylation. Specificity of the IP6 effects on p27 Kip1 and pRb in MCF-7 cells (hormone-dependent) were additionally confirmed in highly invasive hormone-independent MDA-MB 231 breast cancer cells. Use of specific pharmaclogical inhibitors of PKC δ, MEK/Erk, and PI3K/Akt pathways indicated that the IP6-mediated effects on PKC δ were responsible for up-regulation of p27Kip, and pRb hypo-phosphorylation. In addition, IP6-induced apoptosis detected in MCF-7 cells appeared also to be PKC δ-dependent. Our data suggest potential usefulness of IP6 as a novel therapeutic modulator of PKC δ and p27Kip1, an important prognostic factor in human breast cancers.

Keywords

apoptosis breast cancer p27Kip1 phytic acid PKCδ retinoblastoma protein 

Abbreviations

Cdk

cyclin dependent kinase

DMSO

dimethylsulfoxide

DTT

dithiothreitol

EDTA

ethylenediaminetetraacetic acid

Erk

extracellular signal-regulated kinase

ER

estrogen receptor

IP6

inositol hexaphosphate

FCS

fetal calf serum

MAPK

mitogen-activated protein kinase

MEK

MAP kinase kinase

MEM

minimal essential medium

PARP

poly(ADP-ribose) polymerase

PI3-K

phosphatidylinositol 3-kinase

PKB

protein kinase B

PKC

protein kinase C

PMSF

phenylmethylsulfonyl fluoride

pRb

retinoblastoma protein

SDS-PAGE

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

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Copyright information

© Springer 2005

Authors and Affiliations

  • Ivana Vucenik
    • 1
  • Gayatri Ramakrishna
    • 2
  • Kwanchanit Tantivejkul
    • 1
  • Lucy M. Anderson
    • 2
  • Danica Ramljak
    • 2
  1. 1.Department of Medical and Research TechnologyUniversity of Maryland School of MedicineBaltimoreUSA
  2. 2.Laboratory of Comparative CarcinogenesisNCI-FCRDCFrederickUSA

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