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Epigenetic silencing of CXCL12 increases the metastatic potential of mammary carcinoma cells

Oncogene volume 27, pages 1461–1471 (2008)Cite this article

Abstract

Expression of the chemokine receptor CXCR4 has been linked with increased metastasis and decreased clinical prognosis in breast cancer. The current paradigm dictates that CXCR4 fosters carcinoma cell metastasis along a chemotactic gradient to organs expressing the ligand CXCL12. The present study asked if alterations in autocrine CXCR4 signaling via dysregulation of CXCL12 in mammary carcinoma cells modulated their metastatic potential. While CXCR4 was consistently detected, expression of CXCL12 characteristic of human mammary epithelium was silenced by promoter hypermethylation in breast cancer cell lines and primary mammary tumors. Stable re-expression of functional CXCL12 in ligand null cells increased orthotopic primary tumor growth in the mammary fat-pad model of tumorigenesis. Those data parallel increased carcinoma cell proliferation measured in vitro with little-to-no-impact on apoptosis. Moreover, re-expression of autocrine CXCL12 markedly reduced metastatic lung invasion assessed using in vivo bioluminescence imaging following tail vein injection. Consistent with those data, decreased metastasis reflected diminished intracellular calcium signaling and chemotactic migration in response to exogenous CXCL12 independent of changes in CXCR4 expression. Together these data suggest that an elevated migratory signaling response to ectopic CXCL12 contributes to the metastatic potential of CXCR4-expressing mammary carcinoma cells, subsequent to epigenetic silencing of autocrine CXCL12.

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References

  • Agace WW, Amara A, Roberts AI, Pablos JL, Thelen S, Uguccioni M et al. (2000). Constitutive expression of stromal derived factor-1 by mucosal epithelia and its role in HIV transmission and propagation. Curr Biol 10: 325–328.

    Article  CAS  Google Scholar 

  • Amara A, Gall SL, Schwartz O, Salamero J, Montes M, Loetscher P et al. (1997). HIV coreceptor downregulation as antiviral principle: SDF-1alpha-dependent internalization of the chemokine receptor CXCR4 contributes to inhibition of HIV replication. J Exp Med 186: 139–146.

    Article  CAS  Google Scholar 

  • Balkwill F . (2004). The significance of cancer cell expression of the chemokine receptor CXCR4. Semin Cancer Biol 14: 171–179.

    Article  CAS  Google Scholar 

  • Baylin SB, Chen WY . (2005). Aberrant gene silencing in tumor progression: implications for control of cancer. Cold Spring Harb Symp Quant Biol 70: 427–433.427–433.

    Article  CAS  Google Scholar 

  • Brauweiler A, Merrell K, Gauld SB, Cambier JC . (2007). Cutting Edge: acute and chronic exposure of immature B cells to antigen leads to impaired homing and SHIP1-dependent reduction in stromal cell-derived factor-1 responsiveness. J Immunol 178: 3353–3357.

    Article  CAS  Google Scholar 

  • Clark SJ, Harrison J, Molloy PL . (1997). Sp1 binding is inhibited by (m)Cp(m)CpG methylation. Gene 195: 67–71.

    Article  CAS  Google Scholar 

  • Dewan MZ, Ahmed S, Iwasaki Y, Ohba K, Toi M, Yamamoto N . (2006). Stromal cell-derived factor-1 and CXCR4 receptor interaction in tumor growth and metastasis of breast cancer. Biomed Pharmacother 60: 273–276.

    Article  CAS  Google Scholar 

  • Ellison G, Klinowska T, Westwood RF, Docter E, French T, Fox JC . (2002). Further evidence to support the melanocytic origin of MDA-MB-435. Mol Pathol 55: 294–299.

    Article  CAS  Google Scholar 

  • Garcia-Moruja C, Alonso-Lobo JM, Rueda P, Torres C, Gonzalez N, Bermejo M et al. (2005). Functional characterization of SDF-1 proximal promoter. J Mol Biol 348: 43–62.

    Article  CAS  Google Scholar 

  • Gazitt Y . (2004). Homing and mobilization of hematopoietic stem cells and hematopoietic cancer cells are mirror image processes, utilizing similar signaling pathways and occurring concurrently: circulating cancer cells constitute an ideal target for concurrent treatment with chemotherapy and antilineage-specific antibodies. Leukemia 18: 1–10.

    Article  CAS  Google Scholar 

  • Heidemann J, Ogawa H, Dwinell MB, Rafiee P, Maaser C, Gockel HR et al. (2003). Angiogenic effects of interleukin 8 (CXCL8) in human intestinal microvascular endothelial cells are mediated by CXCR2. J Biol Chem 278: 8508–8515.

    Article  CAS  Google Scholar 

  • Herman JG, Baylin SB . (2003). Gene silencing in cancer in association with promoter hypermethylation. N Engl J Med 349: 2042–2054.

    Article  CAS  Google Scholar 

  • Hesselgesser J, Liang M, Hoxie J, Greenberg M, Brass LF, Orsini MJ et al. (1998). Identification and characterization of the CXCR4 chemokine receptor in human T cell lines: ligand binding, biological activity, and HIV-1 infectivity. J Immunol 160: 877–883.

    CAS  Google Scholar 

  • Jenkins DE, Hornig YS, Oei Y, Dusich J, Purchio T . (2005). Bioluminescent human breast cancer cell lines that permit rapid and sensitive in vivo detection of mammary tumors and multiple metastases in immune deficient mice. Breast Cancer Res 7: R444–R454.

    Article  CAS  Google Scholar 

  • Kimura R, Nishioka T, Ishida T . (2003). The SDF1-G801A polymorphism is not associated with SDF1 gene expression in Epstein-Barr virus-transformed lymphoblastoid cells. Genes Immun 4: 356–361.

    Article  CAS  Google Scholar 

  • McGarvey KM, Fahrner JA, Greene E, Martens J, Jenuwein T, Baylin SB . (2006). Silenced tumor suppressor genes reactivated by DNA demethylation do not return to a fully euchromatic chromatin state. Cancer Res 66: 3541–3549.

    Article  CAS  Google Scholar 

  • Moyer RA, Wendt MK, Johanesen PA, Turner JR, Dwinell MB . (2007). Rho activation regulates CXCL12 chemokine stimulated actin rearrangement and restitution in model intestinal epithelia. Lab Invest 87: 807–817.

    Article  CAS  Google Scholar 

  • Muller A, Homey B, Soto H, Ge N, Catron D, Buchanan ME et al. (2001). Involvement of chemokine receptors in breast cancer metastasis. Nature 410: 50–56.

    Article  CAS  Google Scholar 

  • Nagasawa T, Hirota S, Tachibana K, Takakura N, Nishikawa S, Kitamura Y et al. (1996a). Defects of B-cell lymphopoiesis and bone-marrow myelopoiesis in mice lacking the CXC chemokine PBSF/SDF-1. Nature 382: 635–638.

    Article  CAS  Google Scholar 

  • Nagasawa T, Nakajima T, Tachibana K, Iizasa H, Bleul CC, Yoshie O et al. (1996b). Molecular cloning and characterization of a murine pre-B-cell growth-stimulating factor/stromal cell-derived factor 1 receptor, a murine homolog of the human immunodeficiency virus 1 entry coreceptor fusin. Proc Natl Acad Sci USA 93: 14726–14729.

    Article  CAS  Google Scholar 

  • Orimo A, Gupta PB, Sgroi DC, Arenzana-Seisdedos F, Delaunay T, Naeem R et al. (2005). Stromal fibroblasts present in invasive human breast carcinomas promote tumor growth and angiogenesis through elevated SDF-1/CXCL12 secretion. Cell 121: 335–348.

    Article  CAS  Google Scholar 

  • Phillips RJ, Burdick MD, Lutz M, Belperio JA, Keane MP, Strieter RM . (2003). The stromal derived factor-1/CXCL12-CXC chemokine receptor 4 biological axis in non-small cell lung cancer metastases. Am J Respir Crit Care Med 167: 1676–1686.

    Article  Google Scholar 

  • Rhee I, Bachman KE, Park BH, Jair KW, Yen RW, Schuebel KE et al. (2002). DNMT1 and DNMT3b cooperate to silence genes in human cancer cells. Nature 416: 552–556.

    Article  CAS  Google Scholar 

  • Rot A, von Andrian UH . (2004). Chemokines in innate and adaptive host defense: basic chemokinese grammar for immune cells. Annu Rev Immunol 22: 891–928.

    Article  CAS  Google Scholar 

  • Salvucci O, Bouchard A, Baccarelli A, Deschenes J, Sauter G, Simon R et al. (2006). The role of CXCR4 receptor expression in breast cancer: a large tissue microarray study. Breast Cancer Res Treat 97: 275–283.

    Article  CAS  Google Scholar 

  • Schrader AJ, Lechner O, Templin M, Dittmar KE, Machtens S, Mengel M et al. (2002). CXCR4/CXCL12 expression and signalling in kidney cancer. Br J Cancer 86: 1250–1256.

    Article  CAS  Google Scholar 

  • Shirozu M, Nakano T, Inazawa J, Tashiro K, Tada H, Shinohara T et al. (1995). Structure and chromosomal localization of the human stromal cell-derived factor 1 (SDF1) gene. Genomics 28: 495–500.

    Article  CAS  Google Scholar 

  • Smith JM, Johanesen PA, Wendt MK, Binion DG, Dwinell MB . (2005). CXCL12 activation of CXCR4 regulates mucosal host defense through stimulation of epithelial cell migration and promotion of intestinal barrier integrity. Am J Physiol Gastrointest Liver Physiol 288: G316–G326.

    Article  CAS  Google Scholar 

  • Smith MC, Luker KE, Garbow JR, Prior JL, Jackson E, Piwnica-Worms D et al. (2004). CXCR4 regulates growth of both primary and metastatic breast cancer. Cancer Res 64: 8604–8612.

    Article  CAS  Google Scholar 

  • Tachibana K, Hirota S, Iizasa H, Yoshida H, Kawabata K, Kataoka Y et al. (1998). The chemokine receptor CXCR4 is essential for vascularization of the gastrointestinal tract. Nature 393: 591–594.

    Article  CAS  Google Scholar 

  • Ueda Y, Neel NF, Schutyser E, Raman D, Richmond A . (2006). Deletion of the COOH-terminal domain of CXC chemokine receptor 4 leads to the down-regulation of cell-to-cell contact, enhanced motility and proliferation in breast carcinoma cells. Cancer Res 66: 5665–5675.

    Article  CAS  Google Scholar 

  • Wendt MK, Johanesen PA, Kang-Decker N, Binion DG, Shah V, Dwinell MB . (2006). Silencing of epithelial CXCL12 expression by DNA hypermethylation promotes colonic carcinoma metastasis. Oncogene 25: 4986–4997.

    Article  CAS  Google Scholar 

  • Zeelenberg IS, Ruuls-Van Stalle L, Roos E . (2003). The chemokine receptor CXCR4 is required for outgrowth of colon carcinoma micrometastases. Cancer Res 63: 3833–3839.

    CAS  Google Scholar 

  • Zou YR, Kottmann AH, Kuroda M, Taniuchi I, Littman DR . (1998). Function of the chemokine receptor CXCR4 in haematopoiesis and in cerebellar development. Nature 393: 595–599.

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Dr Richard Komorowski (Department of Pathology, Medical College of Wisconsin) and Dr Eric Luedke for assistance in obtaining and processing primary human mammary carcinoma tissue and Dr Robert Truitt (Cancer Center of the Medical College of Wisconsin and Director of the Biophotonic Imaging Core) for assistance in these studies. These studies were supported in part by grants from the Cancer Center of the Medical College of Wisconsin and the American Cancer Society (IRG-84-004).

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  1. Department of Microbiology and Molecular Genetics, Medical College of Wisconsin, Milwaukee, WI, USA

    M K Wendt, A N Cooper & M B Dwinell

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  1. M K Wendt
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  2. A N Cooper
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  3. M B Dwinell
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Correspondence to M B Dwinell.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Wendt, M., Cooper, A. & Dwinell, M. Epigenetic silencing of CXCL12 increases the metastatic potential of mammary carcinoma cells. Oncogene 27, 1461–1471 (2008). https://doi.org/10.1038/sj.onc.1210751

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