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J. Biol. Chem., Vol. 278, Issue 16, 14487-14497, April 18, 2003
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From the National Center for Cell Science (NCCS), NCCS
Complex, Pune-411 007, India
We have recently reported that
osteopontin (OPN) stimulates tumor growth and activation of promatrix
metalloproteinase-2 (pro-MMP-2) through nuclear factor
Osteopontin Induces Nuclear Factor
B-mediated
Promatrix Metalloproteinase-2 Activation through IB
/IKK Signaling
Pathways, and Curcumin (Diferulolylmethane) Down-regulates These
Pathways*
B
(NFB)-mediated induction of membrane type 1 matrix metalloproteinase
(MT1-MMP) in murine melanoma cells (Philip, S., Bulbule, A., and Kundu,
G. C. (2001) J. Biol. Chem. 276, 44926-44935).
However, the molecular mechanism by which OPN activates NFB and
regulates pro-MMP-2 activation in murine melanoma (B16F10) cells is not
well defined. We also investigated the mechanism of action of curcumin
(diferulolylmethane) on OPN-induced NFB-mediated activation of
pro-MMP-2 in B16F10 cells. Here we report that OPN induces
phosphorylation and degradation of the inhibitor of nuclear factor B
(IB
) by inducing the activity of IB kinase (IKK) in these
cells. OPN also induces the nuclear accumulation of NFB p65,
NFB-DNA binding, and transactivation. However, curcumin a known
anti-inflammatory and anticarcinogenic agent suppressed OPN-induced
IB phosphorylation and degradation by inhibiting the IKK
activity. Moreover, our data revealed that curcumin inhibited the
OPN-induced translocation of p65, NFB-DNA binding, and NFB transcriptional activity. The OPN-induced pro-MMP-2 activation and
MT1-MMP expression were also drastically reduced by curcumin. Curcumin
also inhibited OPN-induced cell proliferation, cell migration, extracellular matrix invasion, and synergistically induced
apoptotic morphology with OPN in these cells. Most importantly,
curcumin suppressed the OPN-induced tumor growth in nude mice, and the levels of pro-MMP-2 expression and activation in OPN-induced tumor were
inhibited by curcumin. To our knowledge, this is the first report that
OPN induces NFB activity through phosphorylation and degradation of
IB by activating IKK that ultimately triggers the activation of
pro-MMP-2 and further demonstrates that curcumin potently suppresses
OPN-induced cell migration, tumor growth, and NFB-mediated pro-MMP-2
activation by blocking the IKK/IB signaling pathways.
*
This work was supported by funds from the Department of
Biotechnology (to the National Center for Cell Science) and by an extramural fund from the Department of Science and Technology (to
G. C. K.) of the Government of India.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: National Center for
Cell Science, NCCS Complex, Pune-411 007. Tel.: 91-20-5690931 (ext.
203); Fax: 91-20-5692259; E-mail:
gopalkundu@hotmail.com.
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