CD44+/CD24- breast cancer cells exhibit enhanced invasive properties: an early step necessary for metastasis

IntroductionA subpopulation (CD44+/CD24-) of breast cancer cells has been reported to have stem/progenitor cell properties. The aim of this study was to investigate whether this subpopulation of cancer cells has the unique ability to invade, home, and proliferate at sites of metastasis.MethodsCD44 and CD24 expression was determined by flow cytometry. Northern blotting was used to determine the expression of proinvasive and 'bone and lung metastasis signature' genes. A matrigel invasion assay and intracardiac inoculation into nude mice were used to evaluate invasion, and homing and proliferation at sites of metastasis, respectively.ResultsFive among 13 breast cancer cell lines examined (MDA-MB-231, MDA-MB-436, Hs578T, SUM1315, and HBL-100) contained a higher percentage (>30%) of CD44+/CD24- cells. Cell lines with high CD44+/CD24- cell numbers express basal/mesenchymal or myoepithelial but not luminal markers. Expression levels of proinvasive genes (IL-1α, IL-6, IL-8, and urokinase plasminogen activator [UPA]) were higher in cell lines with a significant CD44+/CD24- population than in other cell lines. Among the CD44+/CD24--positive cell lines, MDA-MB-231 has the unique property of expressing a broad range of genes that favor bone and lung metastasis. Consistent with previous studies in nude mice, cell lines with CD44+/CD24- subpopulation were more invasive than other cell lines. However, only a subset of CD44+/CD24--positive cell lines was able to home and proliferate in lungs.ConclusionBreast cancer cells with CD44+/CD24- subpopulation express higher levels of proinvasive genes and have highly invasive properties. However, this phenotype is not sufficient to predict capacity for pulmonary metastasis.

[1]  René Bernards,et al.  A progression puzzle. , 2002, Nature.

[2]  M. Lohuizen,et al.  Stem Cells and Cancer The Polycomb Connection , 2004, Cell.

[3]  Peter Altevogt,et al.  CD24 affects CXCR4 function in pre-B lymphocytes and breast carcinoma cells , 2006, Journal of Cell Science.

[4]  C. Cordon-Cardo,et al.  A multigenic program mediating breast cancer metastasis to bone. , 2003, Cancer cell.

[5]  Fariba Behbod,et al.  Will cancer stem cells provide new therapeutic targets? , 2005, Carcinogenesis.

[6]  J. López,et al.  CD44 attenuates metastatic invasion during breast cancer progression. , 2005, Cancer research.

[7]  R. Tibshirani,et al.  Gene expression patterns of breast carcinomas distinguish tumor subclasses with clinical implications , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[8]  Ruud H. Brakenhoff,et al.  Dissecting the metastatic cascade , 2004, Nature Reviews Cancer.

[9]  M. Goodell,et al.  A distinct "side population" of cells with high drug efflux capacity in human tumor cells. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[10]  Jean-Michel,et al.  SPARC/osteonectin induces matrix metalloproteinase 2 activation in human breast cancer cell lines. , 1998, Cancer research.

[11]  Yudong D. He,et al.  Gene expression profiling predicts clinical outcome of breast cancer , 2002, Nature.

[12]  H. Nakshatri,et al.  NF-kappaB activation and interleukin 6 production in fibroblasts by estrogen receptor-negative breast cancer cell-derived interleukin 1alpha. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[13]  L. Suva,et al.  Expression of interleukin 8 and not parathyroid hormone-related protein by human breast cancer cells correlates with bone metastasis in vivo. , 2002, Cancer research.

[14]  J. Price.  Metastasis from human breast cancer cell lines , 2005, Breast Cancer Research and Treatment.

[15]  Robert Clarke,et al.  Association of increased basement membrane invasiveness with absence of estrogen receptor and expression of vimentin in human breast cancer cell lines , 1992, Journal of cellular physiology.

[16]  L. Matrisian,et al.  Changing views of the role of matrix metalloproteinases in metastasis. , 1997, Journal of the National Cancer Institute.

[17]  I. Mackenzie,et al.  Retention of intrinsic stem cell hierarchies in carcinoma-derived cell lines. , 2005, Cancer research.

[18]  G. Dontu,et al.  Breast cancer, stem/progenitor cells and the estrogen receptor , 2004, Trends in Endocrinology & Metabolism.

[19]  H. Nakshatri,et al.  NF-κB activation and interleukin 6 production in fibroblasts by estrogen receptor-negative breast cancer cell-derived interleukin 1α , 1998 .

[20]  C. Arteaga,et al.  Targeting the TGF beta signaling network in human neoplasia. , 2003, Cancer cell.

[21]  Charlotte Kuperwasser,et al.  A mouse model of human breast cancer metastasis to human bone. , 2005, Cancer research.

[22]  Carlos L. Arteaga,et al.  Targeting the TGFβ signaling network in human neoplasia , 2003 .

[23]  PAUL A. W. Edwards.  Metastasis: the role of chance in malignancy , 2002, Nature.

[24]  W. Gerald,et al.  Distinct organ-specific metastatic potential of individual breast cancer cells and primary tumors. , 2005, The Journal of clinical investigation.

[25]  S. M. Sims,et al.  Regulation of cancer cell migration and bone metastasis by RANKL , 2006, Nature.

[26]  A. Ouhtit,et al.  Cortactin underpins CD44-promoted invasion and adhesion of breast cancer cells to bone marrow endothelial cells , 2006, Oncogene.

[27]  Paul J. Williams,et al.  Tumor-derived platelet-derived growth factor-BB plays a critical role in osteosclerotic bone metastasis in an animal model of human breast cancer. , 2002, Cancer research.

[28]  Robert M. Hoffman,et al.  Orthotopic Metastatic Mouse Models for Anticancer Drug Discovery and Evaluation: a Bridge to the Clinic , 2004, Investigational New Drugs.

[29]  T. Rowlands,et al.  Cadherins and catenins in breast cancer. , 2005, Current opinion in cell biology.

[30]  J. Sleeman,et al.  CD24 expression causes the acquisition of multiple cellular properties associated with tumor growth and metastasis. , 2005, Cancer research.

[31]  Andy J. Minn,et al.  Genes that mediate breast cancer metastasis to lung , 2005, Nature.

[32]  H. Nakshatri,et al.  NF-κ B Promotes Breast Cancer Cell Migration and Metastasis by Inducing the Expression of the Chemokine Receptor CXCR4* , 2003, Journal of Biological Chemistry.

[33]  François Vaillant,et al.  Generation of a functional mammary gland from a single stem cell , 2006, Nature.

[34]  A. Ashworth,et al.  CD24 staining of mouse mammary gland cells defines luminal epithelial, myoepithelial/basal and non-epithelial cells , 2005, Breast Cancer Research.

[35]  Michael Dean,et al.  Tumour stem cells and drug resistance , 2005, Nature Reviews Cancer.

[36]  R. Walker,et al.  Breast cell invasive potential relates to the myoepithelial phenotype , 2003, International journal of cancer.

[37]  Danila Coradini,et al.  Isolation and in vitro propagation of tumorigenic breast cancer cells with stem/progenitor cell properties. , 2005, Cancer research.

[38]  F. Bertucci,et al.  Gene expression profiling of breast cell lines identifies potential new basal markers , 2006, Oncogene.

[39]  E. Gaffney.  A cell line (HBL-100) established from human breast milk , 2004, Cell and Tissue Research.

[40]  A. Hollestelle,et al.  BRCA1 mutation analysis of 41 human breast cancer cell lines reveals three new deleterious mutants. , 2006, Cancer research.

[41]  L. Bourguignon.  CD44-Mediated Oncogenic Signaling and Cytoskeleton Activation During Mammary Tumor Progression , 2001, Journal of Mammary Gland Biology and Neoplasia.

[42]  Alan Ashworth,et al.  Stem cells and breast cancer: A field in transit , 2003, Nature Reviews Cancer.

[43]  P. Dirks,et al.  Cancer stem cells in nervous system tumors , 2004, Oncogene.

[44]  C. Jordan,et al.  Mechanisms controlling pathogenesis and survival of leukemic stem cells , 2004, Oncogene.

[45]  Christian A. Rees,et al.  Molecular portraits of human breast tumours , 2000, Nature.

[46]  S. Morrison,et al.  Prospective identification of tumorigenic breast cancer cells , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[47]  J. Foidart,et al.  Vimentin contributes to human mammary epithelial cell migration. , 1999, Journal of cell science.

[48]  D. Edwards,et al.  Cancer: Proteases — invasion and more , 1998, Nature.

[49]  I. Weissman,et al.  Stem cells, cancer, and cancer stem cells , 2001, Nature.

[50]  J. Iglehart,et al.  Morphological and biochemical properties of a new human breast cancer cell line. , 1979, Cancer Research.

[51]  Mary Anne Wheeler,et al.  Stem , 1985 .

[52]  Maria Athelogou,et al.  Prevalence of CD44+/CD24-/low cells in breast cancer may not be associated with clinical outcome but may favor distant metastasis. , 2005, Clinical cancer research : an official journal of the American Association for Cancer Research.

[53]  G. Mundy.  Metastasis: Metastasis to bone: causes, consequences and therapeutic opportunities , 2002, Nature Reviews Cancer.