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Growth of hormone-dependent MCF-7 breast cancer cells is promoted by constitutive caveolin-1 whose expression is lost in an EGF-R-mediated manner during development of tamoxifen resistance

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Abstract

Caveolin-1 displays both tumour-suppressor and tumour-promoter properties in breast cancer. Using characterised preclinical cell models for the transition of oestrogen-sensitive (WT-MCF-7 cells) to a tamoxifen-resistant (TAM-R cells) phenotype we examined the role caveolin-1 in the development of hormone-resistant breast cancer. The WT-MCF-7 cells showed abundant expression of caveolin-1 which potentiated oestrogen-receptor (ERα) signalling and promoted cell growth despite caveolin-1 mediating inhibition of ERK signalling. In TAM-R cells caveolin-1 expression was negligible, repressed by EGF-R/ERK signalling. Pharmacological inhibition of EGFR/ERK in TAM-R cells restored caveolin-1 and also resulted in the emergence of pools of phosphorylated caveolin-1. WT-MCF-7 cells exposed to tamoxifen for upto 12 weeks displayed increased caveolin-1 (peaking by week 2) followed (after week 8) by a marked decrease as the cells progress to develop a stable tamoxifen-resistant phenotype. The targeted down-regulation (siRNA) of caveolin-1 in WT-MCF-7 cells reduced growth but did not affect their sensitivity to tamoxifen, suggesting loss of caveolin-1 alone is not sufficient to confer tamoxifen-resistance. Hyperactivation of EGFR/ERK is a feature of tamoxifen-resistant breast cancer cells, a principal driver of cell growth. Recombinant expression of caveolin-1 in TAM-R cells did not affect EGFR/ERK activity, potentially due to mislocalisation of caveolin-1 through hyperactivation of the mTOR pathway or altered caveolin-1 phosphorylation. This work defines a novel role for caveolin-1 with implications for the clinical course of breast cancer and identifies caveolin-1 as a potential drug target for the treatment of early oestrogen-dependent breast cancers. Further, the loss of caveolin-1 may have benefit as a molecular signature for tamoxifen resistance.

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References

  1. Early Breast Cancer Trialists’ Collaborative Group (EBCTCG) (2005) Effects of chemotherapy and hormonal therapy for early breast cancer on recurrence and 15-year survival: an overview of the randomised trails. Lancet 365:1687–1717. doi:10.1016/S0140-6736(05)66544-0

    Article  CAS  Google Scholar 

  2. Powles T, Ashley S, Tidy A et al (2000) Twenty-year follow-up of the Royal Marsden randomized, double-blinded tamoxifen breast cancer prevention trial. J Natl Cancer Inst 99:283–290. doi:10.1093/jnci/djk050

    Google Scholar 

  3. Clarke R, Leonessa F, Welch J et al (2001) Cellular and molecular pharmacology of antiestrogen action and resistance. Pharmacol Rev 53:25–71

    PubMed  CAS  Google Scholar 

  4. Nicholson R, Gee J (2000) Oestrogen and growth factor cross-talk and endocrine insensitivity and acquired resistance in breast cancer. Br J Cancer 82:501–513. doi:10.1054/bjoc.1999.0954

    Article  PubMed  CAS  Google Scholar 

  5. Nicholson R, Hutcheson I, Hiscox S et al (2005) Growth factor signalling and resistance to selective oestrogen receptor modulators and pure anti-oestrogens: the use of anti-growth factor therapies to treat or delay endocrine resistance in breast cancer. Endocr Relat Cancer 12:S29–S36. doi:10.1677/erc.1.00991

    Article  PubMed  CAS  Google Scholar 

  6. Long B, McKibben B, Lynch M, van den Berg H (1992) Changes in epidermal growth factor receptor expression and response to ligand associated with acquired tamoxifen resistance or oestrogen independence in the ZR-75–1 human breast cancer cell line. Br J Cancer 65:865–869

    PubMed  CAS  Google Scholar 

  7. Coutts A, Murphy L (1998) Elevated mitogen-activated protein kinase activity in estrogen-nonresponsive human breast cancer cells. Cancer Res 58:4071–4074

    PubMed  CAS  Google Scholar 

  8. El-Ashry D, Miller D, Kharbanda S et al (1997) Constituitive Raf-1 kinase activity in breast cancer cells induces both estrogen-independent growth and apoptosis. Oncogene 15:423–435. doi:10.1038/sj.onc.1201198

    Article  PubMed  CAS  Google Scholar 

  9. Gee J, Robertson J, Ellis I et al (2001) Phosphorylation of ERK1/2 mitogen-activated protein kinase is associated with poor response to anti-hormonal therapy and decreased patient survival in clinical breast cancer. Int J Cancer 95:247–254. doi:10.1002/1097-0215(20010720)95:4≤247::AID-IJC1042≥3.0.CO;2-S

    Article  PubMed  CAS  Google Scholar 

  10. McClelland R, Barrow D, Madden T et al (2001) Enhanced epidermal growth factor receptor signalling in MCF7 breast cancer cells following long-term culture in the presence of the pure antioestrogen ICI 182, 780 (Faslodex). Endocrinology 142:2776–2788. doi:10.1210/en.142.7.2776

    Article  PubMed  CAS  Google Scholar 

  11. Knowlden J, Hutcheson I, Jones H et al (2003) Elevated levels of epidermal growth receptor/c-erB2 heterodimers mediate an autocrine growth regulatory pathway in tamoxifen-resistant MCF-7 cells. Endocrinology 144:1032–1044. doi:10.1210/en.2002-220620

    Article  PubMed  CAS  Google Scholar 

  12. Williams T, Lisanti M (2004) The caveolin genes: from cell biology to medicine. Ann Med 36:584–595. doi:10.1080/07853890410018899

    Article  PubMed  CAS  Google Scholar 

  13. Couet J, Li S, Okamoto T et al (1997) Identification of peptide and protein ligands for the caveolin-scaffolding domain. Implications for the interaction of caveolin with caveolae-associated proteins. J Biol Chem 272:6525–6533. doi:10.1074/jbc.272.48.30429

    Article  PubMed  CAS  Google Scholar 

  14. Li S, Couet J, Lisanti M (1996) Src tyrosine kinases, Galpha subunits, and H-Ras share a common membrane-anchored scaffolding domain, caveolin. Caveolin binding negatively regulates the auto-activation of Src tyrosine kinases. J Biol Chem 271:29182–29190. doi:10.1074/jbc.271.46.29182

    Article  PubMed  CAS  Google Scholar 

  15. Li S, Okamoto T, Chun M et al (1995) Evidence for a regulated interaction between heterotrimeric G proteins and caveolin. J Biol Chem 270:15693–15701. doi:10.1074/jbc.270.26.15693

    Article  PubMed  CAS  Google Scholar 

  16. Couet J, Sargiacomo M, Lisanti MP (1997) Interaction of a receptor tyrosine kinase, EGF-R, with caveolins. Caveolin binding negatively regulates tyrosine and serine/threonine kinase activities. J Biol Chem 272:30429–30438. doi:10.1074/jbc.272.48.30429

    Article  PubMed  CAS  Google Scholar 

  17. Engelman J, Chu C, Lin A et al (1998) Caveolin-mediated regulation of signalling along the p42/p44 MAP kinase cascade in vivo. FEBS Lett 428:205–211. doi:10.1016/S0014-5793(98)00470-0

    Article  PubMed  CAS  Google Scholar 

  18. Engelman J, Lee R, Karnezis A et al (1998) Reciprocal regulation of neu tyrosine kinase activity and caveolin-1 protein expression in vitro and in vivo. Implications for human breast cancer. J Biol Chem 273:20448–20455. doi:10.1074/jbc.273.32.20448

    Article  PubMed  CAS  Google Scholar 

  19. Engelman J, Zhang X, Razani B et al (1999) p42/44 MAP kinase-dependent and -independent signaling pathways regulate caveolin-1 gene expression. Activation of Ras-MAP kinase and protein kinase a signaling cascades transcriptionally down-regulates caveolin-1 promoter activity. J Biol Chem 274:32333–32341. doi:10.1074/jbc.274.45.32333

    Article  PubMed  CAS  Google Scholar 

  20. Lin M, DiVito M, Merajver S et al (2005) Regulation of pancreatic cancer cell migration and invasion by RhoC GTP ase and caveolin-1. Mol Cancer 4:21. doi:10.1186/1476-4598-4-21

    Article  PubMed  CAS  Google Scholar 

  21. Lee S, Reimer C, Oh P et al (1998) Tumor cell growth inhibition by caveolin re-expression in human breast cancer cells. Oncogene 16:1391–1397. doi:10.1038/sj.onc.1201661

    Article  PubMed  CAS  Google Scholar 

  22. Suzuki T, Suzuki Y, Hanada K et al (1998) Reduction of caveolin-1 in tumorigenic human cell hybrids. J Biochem 124:383–388

    PubMed  CAS  Google Scholar 

  23. Zou W, McDaneld L, Smith L (2003) Caveolin-1 haploinsufficiency leads to partial transformation of human breast epithelial cells. Anticancer Res 23:4581–4586

    PubMed  CAS  Google Scholar 

  24. Fiucci G, Ravid D, Reich R et al (2002) Caveolin-1 inhibits anchorage-independent growth, ankoikis and invasiveness in MCF-7 human breast cancer cells. Oncogene 21:2365–2375. doi:10.1038/sj.onc.1205300

    Article  PubMed  CAS  Google Scholar 

  25. Engelman J, Zang X, Lisanti M (1998) Genes encodoing human caveolin-1 and -2 are co-localized to the D7S522 locus (7q31.1), a known fragile site (FRA7G) that is frequently deleted in human cancers. FEBS Lett 436:403–410. doi:10.1016/S0014-5793(98)01134-X

    Article  PubMed  CAS  Google Scholar 

  26. Schlegel A, Wang C, Katzenellenbogen B et al (1999) Caveolin-1 potentiates estrogen receptor alpha (ERalpha) signaling. Caveolin-1 drives ligand-independent nuclear translocation and activation of ERalpha. J Biol Chem 274:33551–33556. doi:10.1074/jbc.274.47.33551

    Article  PubMed  CAS  Google Scholar 

  27. Schlegel A, Wang C, Pestell R et al (2002) Ligand-independent activation of oestrogen receptor alpha by caveolin-1. Biochem J 359:203–210. doi:10.1042/0264-6021:3590203

    Article  Google Scholar 

  28. Nasu Y, Timme T, Yang G et al (1998) Suppression of caveolin expression induces androgen sensitivity in metastatic androgen insensitive mouse prostate cancer cell lines. Nat Med 4:1062–1064. doi:10.1038/2048

    Article  PubMed  CAS  Google Scholar 

  29. Yang G, Truong L, Wheeler T et al (1999) Caveolin-1 expression in clinically confined human prostate cancer: a novel prognostic marker. Cancer Res 59:5719–5723

    PubMed  CAS  Google Scholar 

  30. Rau K, Kang H, Cha T et al (2005) The mechanisms and managements of hormone therapy resistance in breast and prostate cancers. Endocr Relat Cancer 12:511–532. doi:10.1677/erc.1.01026

    Article  PubMed  CAS  Google Scholar 

  31. Hutcheson I, Knowlden J, Madden T et al (2003) Oestrogen receptor-mediated modulation of the EGFR/MAPK pathway in tamoxifen-resistant MCF-7 cells. Breast Cancer Res Treat 81:81–93. doi:10.1023/A:1025484908380

    Article  PubMed  CAS  Google Scholar 

  32. Cho K, Ryu S, Oh Y et al (2004) Morphological adjustment of senescent cells by modulating caveolin-1 status. J Biol Chem 279:42270–42278. doi:10.1074/jbc.M402352200

    Article  PubMed  CAS  Google Scholar 

  33. Shin J, Kim J, Ryu B et al (2006) Caveolin-1 is associated with VCAM-1 dependent adhesion of gastric cancer cells to endothelial cells. Cell Physiol Biochem 17:211–220. doi:10.1159/000094126

    Article  PubMed  CAS  Google Scholar 

  34. Song K, Li S, Okamoto T et al (1996) Co-purification and direct interaction of Ras with caveolin, an integral membrane protein of caveolae microdomains. Detergent-free purification of caveolae microdomains. J Biol Chem 271:9690–9697. doi:10.1074/jbc.271.16.9690

    Article  PubMed  CAS  Google Scholar 

  35. Williams T, Lisanti M (2005) Caveolin-1 in oncogenic transformation, cancer, and metastasis. Am J Cell Physiol 288:C494–C506. doi:10.1152/ajpcell.00458.2004

    Article  CAS  Google Scholar 

  36. Goetz J, Lajoie P, Wiseman S et al (2008) Caveolin-1 in tumour progression: the good, the bad and the ugly. Cancer Metastasis Rev 27:715–735. doi:10.1007/s10555-008-9160-9

    Article  PubMed  CAS  Google Scholar 

  37. Burgermeister E, Liscovitch M, Rocken C et al (2008) Caveats of caveolin-1 in cancer progression. Cancer Lett 268:187–201. doi:10.1016/j.canlet.2008.03.055

    Article  PubMed  CAS  Google Scholar 

  38. Zhang W, Razani B, Altschuler Y et al (2000) Caveolin-1 inhibits epidermal growth factor-stimulated lamellipod extension and cell migration in metastatic mammary adenocarcinoma cells (MTLn3). J Biol Chem 275:20717–20725. doi:10.1074/jbc.M909895199

    Article  PubMed  CAS  Google Scholar 

  39. Park D, Hyangkyu L, Frank P et al (2002) Caveolin-1-deficient mice show accelerated mammary gland development during pregnancy, premature lactation, and hyperactivation of the Jak-2/STAT5a signalling cascade. Mol Biol Cell 13:3416–3430. doi:10.1091/mbc.02-05-0071

    Article  PubMed  CAS  Google Scholar 

  40. Williams T, Medina F, Badano I et al (2004) Caveolin-1 gene promotes mammary tumorigenesis and dramatically enhances lung metastasis in vivo. J Biol Chem 279:51630–51646. doi:10.1074/jbc.M409214200

    Article  PubMed  CAS  Google Scholar 

  41. Williams T, Sotgia F, Lee H et al (2006) Stromal and epithelial caveolin-1 both confer a protective effect against mammary hyperplasia and tumorigenesis. Am J Pathol 169:1784–1801. doi:10.2353/ajpath.2006.060590

    Article  PubMed  CAS  Google Scholar 

  42. Hayashi K, Matsuda S, Machida K et al (2001) Invasion activating caveolin-1 mutation in human scirrhous breast cancers. Cancer Res 61:2361–2364

    PubMed  CAS  Google Scholar 

  43. Pinilla S, Honrado E, Hardisson E et al (2006) Caveolin-1 expression is associated with a basal-like phenotype in sporadic and hereditary breast cancer. Breast Cancer Res Treat 99:85–90. doi:10.1007/s10549-006-9184-1

    Article  PubMed  CAS  Google Scholar 

  44. Perrone G, Altomare V, Zagami M et al (2009) Caveolin-1 expression in human breast lobular cancer progression. Mod Pathol 22:71–78. doi:10.1038/modpathol.2008.154

    Article  PubMed  CAS  Google Scholar 

  45. Zschocke J, Manthey D, Bayatti N et al (2002) Estrogen receptor alpha-mediated silencing of caveolin gene expression in neuronal cells. J Biol Chem 277:38772–38780. doi:10.1074/jbc.M205664200

    Article  PubMed  CAS  Google Scholar 

  46. Razandi M, Oh P, Pedram A et al (2002) ERs associate with and regulate the production of caveolin: implications for signaling and cellular actions. Mol Endocrinol 16:100–115. doi:10.1210/me.16.1.100

    Article  PubMed  CAS  Google Scholar 

  47. Li T, Sotgia F, Vuolo M et al (2006) Caveolin-1 mutations in human breast cancer: functional association with estrogen receptor alpha-positive status. Am J Pathol 168:1998–2013. doi:10.2353/ajpath.2006.051089

    Article  PubMed  CAS  Google Scholar 

  48. Park S, Kim J, Kim Y (2005) Caveolin-1 is down-regulated and inversely correlated with HER2 and EGFR expression status in invasive ductal carcinoma of the breast. Histopathol 47:625–630. doi:10.1111/j.1365-2559.2005.02303.x

    Article  Google Scholar 

  49. Sagara Y, Mimori K, Yoshinaga K et al (2004) Clinical significance of Caveolin-1, Caveolin-2 and HER2/neu mRNA expression in human breast cancer. Br J Cancer 91:959–965

    PubMed  CAS  Google Scholar 

  50. Britton D, Hutcheson I, Knowlden J et al (2006) Bidirectional cross talk between ERα and EGFR signalling pathways regulates tamoxifen-resistant growth. Breast Cancer Res Treat 96:131–146. doi:10.1007/s10549-005-9070-2

    Article  PubMed  CAS  Google Scholar 

  51. Jones K, Jiang X, Yamamoto Y et al (2004) Tuberin is a component of lipid rafts and mediates caveolin-1 localisation: role of TSC2 in post-Golgi transport. Exp Cell Res 295:512–524. doi:10.1016/j.yexcr.2004.01.022

    Article  PubMed  CAS  Google Scholar 

  52. Jiang X, Yeung S (2006) Regulation of microtubule-dependent protein transport by the TSC2/mammalian target of rapamycin pathway. Cancer Res 66:2006

    Google Scholar 

  53. Aoki T, Nomura R, Fujimoto T (1999) Tyrosine phosphorylation of caveolin-1 in the endothelium. Exp Cell Res 253:629–636. doi:10.1006/excr.1999.4652

    Article  PubMed  CAS  Google Scholar 

  54. Nomura R, Fujimoto T (1999) Tyrosine-phosphorylated caveolin-1: immunolocalization and molecular characterization. Mol Biol Cell 10:975–986

    PubMed  CAS  Google Scholar 

  55. Li S, Seitz R, Lisanti M (1996) Phosphorylation of caveolin by src tyrosine kinases. The alpha-isoform of caveolin is selectively phosphorylated by v-Src in vivo. J Biol Chem 271:3863–3868. doi:10.1074/jbc.271.7.3863

    Article  PubMed  CAS  Google Scholar 

  56. Hiscox S, Morgan L, Green T et al (2006) Elevated Src activity promotes cellular invasion and motility in tamoxifen resistant breast cancer cells. Breast Cancer Res Treat 97:263–274. doi:10.1007/s10549-005-9120-9

    Article  PubMed  CAS  Google Scholar 

  57. Lee H, Volonte D, Galbiati F et al (2000) Constitutive and growth factor-regulated phosphorylation of caveolin-1 occurs at the same site (Tyr-14) in vivo: identification of a c-Src/Cav-1/Grb7 signaling cassette. Mol Endocrinol 14:1750–17575. doi:10.1210/me.14.11.1750

    Article  PubMed  CAS  Google Scholar 

  58. Khan EM, Heidinger JM, Levy M et al (2006) Epidermal growth factor receptor exposed to oxidative stress undergoes Src- and caveolin-1-dependent perinuclear trafficking. J Biol Chem 281:14486–14493. doi:10.1074/jbc.M509332200

    Article  PubMed  CAS  Google Scholar 

  59. Dittman K, Mayer C, Kehlbach R, Rodemann H (2008) The radioprotector Bowman-Birk proteinase inhibitor stimulates DNA repair via the epidermal growth factor receptor phosphorylation and nuclear transport. Radiother Oncol 86:375–382. doi:10.1016/j.radonc.2008.01.007

    Article  CAS  Google Scholar 

  60. Joshi B, Strugnell S, Goetz JG et al (2008) Phosphorylated caveolin-1 regulates Rho/ROCK-dependent focal adhesion dynamics and tumor cell migration and invasion. Cancer Res 68:8210–8220. doi:10.1158/0008-5472.CAN-08-0343

    Article  PubMed  CAS  Google Scholar 

  61. Van den Eynden G, Van Laere S, Van der Auwera I et al (2006) Overexpression of caveolin-1 and -2 in cell lines and in human samples of inflammatory breast cancer. Breast Cancer Res Treat 95:219–228. doi:10.1007/s10549-005-9002-1

    Article  PubMed  CAS  Google Scholar 

  62. Johnston SR (2005) Clinical trials of intracellular signal transductions inhibitors for breast cancer—a strategy to overcome endocrine resistance. Endocr Relat Cancer 1(suppl):S145–S157. doi:10.1677/erc.1.00992

    Article  CAS  Google Scholar 

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Acknowledgments

The authors would like to thank Carol Dutkowski and Denise Barrow for their excellent technical assistance. To Dr. Andrew Hollins for construction of the recombinant caveolin-1 construct and Dr. Arwyn T. Jones for advice on subcellular fractionation. This research was generously supported by the Tenovus organisation and a Welsh School of Pharmacy studentship to NBPT. ‘Iressa’ and ‘Faslodex’ are trademarks of the AstraZeneca group of companies.

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  1. Experimental Therapeutics, Welsh School of Pharmacy, Cardiff University, Redwood Building, , King Edward VII Avenue, Cardiff, CF10 3XF, UK

    Nicholas B. P. Thomas, Lee Campbell & Mark Gumbleton

  2. Tenovus Centre for Cancer Research, Cardiff University, Redwood Building, , King Edward VII Avenue, Cardiff, CF10 3XF, UK

    Nicholas B. P. Thomas, Iain R. Hutcheson, Julia Gee, Kathryn M. Taylor & Robert I. Nicholson

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  1. Nicholas B. P. Thomas
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Correspondence to Mark Gumbleton.

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Nicholas B.·P. Thomas, Iain R. Hutcheson and Lee Campbell have contributed equally to this work.

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Thomas, N.B.P., Hutcheson, I.R., Campbell, L. et al. Growth of hormone-dependent MCF-7 breast cancer cells is promoted by constitutive caveolin-1 whose expression is lost in an EGF-R-mediated manner during development of tamoxifen resistance. Breast Cancer Res Treat 119, 575–591 (2010). https://doi.org/10.1007/s10549-009-0355-8

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