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Originally published In Press as doi:10.1074/jbc.M300609200 on April 9, 2003

J. Biol. Chem., Vol. 278, Issue 24, 21631-21638, June 13, 2003
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NF-{kappa} B Promotes Breast Cancer Cell Migration and Metastasis by Inducing the Expression of the Chemokine Receptor CXCR4*

Gregory Helbig {ddagger} §, Kent W. Christopherson , II § ¶ || ** {ddagger}{ddagger}, Poornima Bhat-Nakshatri || **, Suresh Kumar {ddagger}, Hiromitsu Kishimoto {ddagger}, Kathy D. Miller §§, Hal E. Broxmeyer ¶ || ** §§ and Harikrishna Nakshatri {ddagger} || ** ¶¶ ||||

From the Departments of {ddagger}Surgery, Microbiology and Immunology, §§Medicine, and ¶¶Biochemistry and Molecular Biology and the ||Walther Oncology Center, Indiana University School of Medicine, Indianapolis, Indiana 46202 and the** Walther Cancer Institute, Indianapolis, Indiana 46208

Metastasis of cancer cells is a complex process involving multiple steps including invasion, angiogenesis, and trafficking of cancer cells through blood vessels, extravasations, organ-specific homing, and growth. While matrix metalloproteinases, urokinase-type plasminogen activator, and cytokines play a major role in invasion and angiogenesis, chemokines such as stromal derived factor-1{alpha} (SDF-1{alpha}) and their receptors such as CXCR4 are thought to play a critical role in motility, homing, and proliferation of cancer cells at specific metastatic sites. We and others have previously reported that the extracellular signal-activated transcription factor NF-{kappa}B up-regulates the expression of matrix metalloproteinases, urokinase-type plasminogen activator, and cytokines in highly metastatic breast cancer cell lines. In this report, we demonstrate that NF-{kappa}B regulates the motility of breast cancer cells by directly up-regulating the expression of CXCR4. Overexpression of the inhibitor of {kappa}B (I{kappa}B) in breast cancer cells with constitutive NF-{kappa}B activity resulted in reduced expression of CXCR4 and a corresponding loss of SDF-1{alpha}-mediated migration in vitro. Introduction of CXCR4 cDNA into I{kappa}B-expressing cells restored SDF-1{alpha}-mediated migration. Electrophoretic mobility shift assays and transient transfection assays revealed that the NF-{kappa}B subunits p65 and p50 bind directly to sequences within the –66 to +7 region of the CXCR4 promoter and activate transcription. We also show that the cell surface expression of CXCR4 and the SDF-1{alpha}-mediated migration are enhanced in breast cancer cells isolated from mammary fat pad xenografts compared with parental cells grown in culture. A further increase in CXCR4 cell surface expression and SDF-1{alpha}-mediated migration was observed with cancer cells that metastasized to the lungs. Taken together, these results implicate NF-{kappa}B in the migration and the organ-specific homing of metastatic breast cancer cells.


Received for publication, January 20, 2003 , and in revised form, March 17, 2003.

* This work was supported by American Cancer Society Grant RPG-00-122-01-TBE, American Institute for Cancer Research Grant 00A047, National Institutes of Health Grant CA82208, and the Phi Beta Psi Sorority (all to H. N.) and by National Institutes of Health Grants DK53674 and HL67384 (to H. E. B.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Both authors contributed equally to this work.

{ddagger}{ddagger} Supported by National Institutes of Health T32 Training Program Grant DK07519 (to H. E. B.).

|||| To whom correspondence should be addressed: R4-202 Indiana Cancer Research Inst., 1044 W. Walnut St., Indianapolis, IN 46202. Tel.: 317-278-2238; Fax: 317-274-0396; E-mail: hnakshat{at}iupui.edu.


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F. Pacifico, C. Mauro, C. Barone, E. Crescenzi, S. Mellone, M. Monaco, G. Chiappetta, G. Terrazzano, D. Liguoro, P. Vito, et al.
Oncogenic and Anti-apoptotic Activity of NF-{kappa}B in Human Thyroid Carcinomas
J. Biol. Chem., December 24, 2004; 279(52): 54610 - 54619.
[Abstract] [Full Text] [PDF]


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Cancer Res.Home page
M. C. P. Smith, K. E. Luker, J. R. Garbow, J. L. Prior, E. Jackson, D. Piwnica-Worms, and G. D. Luker
CXCR4 Regulates Growth of Both Primary and Metastatic Breast Cancer
Cancer Res., December 1, 2004; 64(23): 8604 - 8612.
[Abstract] [Full Text] [PDF]


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Mol. Cell. Biol.Home page
Z.-l. Yuan, Y.-j. Guan, L. Wang, W. Wei, A. B. Kane, and Y. E. Chin
Central Role of the Threonine Residue within the p+1 Loop of Receptor Tyrosine Kinase in STAT3 Constitutive Phosphorylation in Metastatic Cancer Cells
Mol. Cell. Biol., November 1, 2004; 24(21): 9390 - 9400.
[Abstract] [Full Text] [PDF]


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J. Virol.Home page
A. Owen, B. Chandler, P. G. Bray, S. A. Ward, C. A. Hart, D. J. Back, and S. H. Khoo
Functional Correlation of P-Glycoprotein Expression and Genotype with Expression of the Human Immunodeficiency Virus Type 1 Coreceptor CXCR4
J. Virol., November 1, 2004; 78(21): 12022 - 12029.
[Abstract] [Full Text] [PDF]


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CarcinogenesisHome page
L. Tacchini, C. De Ponti, E. Matteucci, R. Follis, and M.A. Desiderio
Hepatocyte growth factor-activated NF-{kappa}B regulates HIF-1 activity and ODC expression, implicated in survival, differently in different carcinoma cell lines
Carcinogenesis, November 1, 2004; 25(11): 2089 - 2100.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
M.-T. Lin, C.-C. Chang, S.-T. Chen, H.-L. Chang, J.-L. Su, Y.-P. Chau, and M.-L. Kuo
Cyr61 Expression Confers Resistance to Apoptosis in Breast Cancer MCF-7 Cells by a Mechanism of NF-{kappa}B-dependent XIAP Up-Regulation
J. Biol. Chem., June 4, 2004; 279(23): 24015 - 24023.
[Abstract] [Full Text] [PDF]


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Mol Cancer ResHome page
B.-C. Lee, T.-H. Lee, S. Avraham, and H. K. Avraham
Involvement of the Chemokine Receptor CXCR4 and Its Ligand Stromal Cell-Derived Factor 1{alpha} in Breast Cancer Cell Migration Through Human Brain Microvascular Endothelial Cells
Mol. Cancer Res., June 1, 2004; 2(6): 327 - 338.
[Abstract] [Full Text] [PDF]


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Am. J. Pathol.Home page
S. Kumar, H. Kishimoto, H. L. Chua, S. Badve, K. D. Miller, R. M. Bigsby, and H. Nakshatri
Interleukin-1{alpha} Promotes Tumor Growth and Cachexia in MCF-7 Xenograft Model of Breast Cancer
Am. J. Pathol., December 1, 2003; 163(6): 2531 - 2541.
[Abstract] [Full Text] [PDF]


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Proc. Natl. Acad. Sci. USAHome page
J. B. Rubin, A. L. Kung, R. S. Klein, J. A. Chan, Y. Sun, K. Schmidt, M. W. Kieran, A. D. Luster, and R. A. Segal
A small-molecule antagonist of CXCR4 inhibits intracranial growth of primary brain tumors
PNAS, November 11, 2003; 100(23): 13513 - 13518.
[Abstract] [Full Text] [PDF]




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