Virchows Archiv

, Volume 451, Issue 6, pp 1039–1045

COX-2 localization within plasma membrane caveolae-like structures in human lobular intraepithelial neoplasia of the breast

  • Giuseppe Perrone
  • Mariagiovanna Zagami
  • Vittorio Altomare
  • Cleonice Battista
  • Sergio Morini
  • Carla Rabitti
Original Article


Cyclooxygenase-2 (COX-2) is highly expressed in human intraepithelial neoplasia of the breast and takes part in the molecular pathway implicated in progression of breast cancer. Recently, we demonstrated that COX-2 protein is mainly located in plasma membrane of lobular intraepithelial neoplasia (LIN) cells suggesting a localization in caveolae-like structures. The aim of the present study is to establish subcellular locations of COX-2 and its colocalization with caveolin-1 (CAV-1) to caveolae structures in LIN. To establish a relationship between COX-2 and CAV-1, 39 LINs were studied by immunohistochemistry and confocal microscopy analysis. COX-2 and CAV-1 expression was observed respectively in 79.5 and in 94.9% of LIN studied. A positive correlation was found between membrane COX-2 staining pattern and CAV-1 expression, while no correlation was found between cytoplasm COX-2 staining pattern and CAV-1. Confocal analysis showed that COX-2 localized to plasma membrane was strictly associated to CAV-1 suggesting that an amount of COX-2 protein is placed in caveolae-like structures. Our results show that COX-2 is localized within caveolae compartment and colocalized with CAV-1 protein in LIN lesions. Because caveolae are rich in signaling molecules, this COX-2 compartment may play an important role in diverse breast cancer carcinogenesis processes.


Lobular intraepithelial neoplasia CAV-1 COX-2 Immunohistochemistry Confocal microscopy 


  1. 1.
    Balsinde J, Balboa MA, Dennis EA (1998) Functional coupling between secretory phospholipase A2 and cyclooxygenase-2 and its regulation by cytosolic group IV phospholipase A2. Proc Natl Acad Sci U S A 95:7951–7956PubMedCrossRefGoogle Scholar
  2. 2.
    Couet J, Li S, Okamoto T, Ikezu T, Lisanti MP (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 272:6525–6533PubMedCrossRefGoogle Scholar
  3. 3.
    Engelman JA, Lee RJ, Karnezis A, Bearss DJ, Webster M, Siegel P, Muller WJ, Windle JJ, Pestell RG, Lisanti MP (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 273:20448–20455PubMedCrossRefGoogle Scholar
  4. 4.
    Esaki M, Matsumoto T, Yao S, Nakamura S, Hirahashi M, Yao T, Iida M (2005) Immunohistochemical characteristics of duodenal adenomas in familial adenomatous polyposis with special reference to cell kinetics. Human Pathol 36:66–73CrossRefGoogle Scholar
  5. 5.
    Half E, Tang XM, Gwyn K, Sahin A, Wathen K, Sinicrope FA (2002) Cyclooxygenase-2 expression in human breast cancers and adjacent ductal carcinoma in situ. Cancer Res 62:1676–1681PubMedGoogle Scholar
  6. 6.
    Harris RE, Beebe-Donk J, Alshafie GA (2006) Reduction in the risk of human breast cancer by selective cyclooxygenase-2 (COX-2) inhibitors. BMC Cancer 6:27PubMedCrossRefGoogle Scholar
  7. 7.
    Hurlstone AF, Reid G, Reeves JR, Fraser J, Strathdee G, Rahilly M, Parkinson EK, Black DM (1999) Analysis of the CAVEOLIN-1 gene at human chromosome 7q31.1 in primary tumours and tumour-derived cell lines. Oncogene 18:1881–1890PubMedCrossRefGoogle Scholar
  8. 8.
    Kargman SL, O’Neill GP, Vickers PJ, Evans JF, Mancini JA, Jothy S (1995) Expression of prostaglandin G/H synthase-1 and -2 protein in human colon cancer. Cancer Res 55:2556–2559PubMedGoogle Scholar
  9. 9.
    Konturek PC, Kania J, Burnat G, Hahn EG, Konturek SJ (2005) Prostaglandins as mediators of COX-2 derived carcinogenesis in gastrointestinal tract. J Physiol Pharmacol 56(Suppl 5):57–73PubMedGoogle Scholar
  10. 10.
    Kurumbail RG, Stevens AM, Gierse JK, McDonald JJ, Stegeman RA, Pak JY, Gildehaus D, Miyashiro JM, Penning TD, Seibert K, Isakson PC, Stallings WC (1996) Structural basis for selective inhibition of cyclooxygenase-2 by anti-inflammatory agents. Nature 384:644–648PubMedCrossRefGoogle Scholar
  11. 11.
    Kuwata H, Sawada H, Murakami M, Kudo I (1999) Role of type IIA secretory phospholipase A2 in arachidonic acid metabolism. Adv Exp Med Biol 469:183–188PubMedGoogle Scholar
  12. 12.
    Lee SW, Reimer CL, Oh P, Campbell DB, Schnitzer JE (1998) Tumor cell growth inhibition by caveolin re-expression in human breast cancer cells. Oncogene 16:1391–1397PubMedCrossRefGoogle Scholar
  13. 13.
    Li S, Couet J, Lisanti MP (1996) Src tyrosine kinases, Galpha subunits, and H-Ras share a common membrane-anchored scaffolding protein, caveolin. Caveolin binding negatively regulates the auto-activation of Src tyrosine kinases. J Biol Chem 271:29182–29190PubMedCrossRefGoogle Scholar
  14. 14.
    Liou JY, Shyue SK, Tsai MJ, Chung CL, Chu KY, Wu KK (2000) Colocalization of prostacyclin synthase with prostaglandin H synthase-1 (PGHS-1) but not phorbol ester-induced PGHS-2 in cultured endothelial cells. J Biol Chem 275:15314–15320PubMedCrossRefGoogle Scholar
  15. 15.
    Liou JY, Deng WG, Gilroy DW, Shyue SK, Wu KK (2001) Colocalization and interaction of cyclooxygenase-2 with caveolin-1 in human fibroblasts. J Biol Chem 276:34975–34982PubMedCrossRefGoogle Scholar
  16. 16.
    Liu CH, Chang SH, Narko K, Trifan OC, Wu MT, Smith E, Haudenschild C, Lane TF, Hla T (2001) Overexpression of cyclooxygenase-2 is sufficient to induce tumorigenesis in transgenic mice. J Biol Chem 276:18563–18569PubMedCrossRefGoogle Scholar
  17. 17.
    Liu J, Oh P, Horner T, Rogers RA, Schnitzer JE (1997) Organized endothelial cell surface signal transduction in caveolae distinct from glycosylphosphatidylinositol-anchored protein microdomains. J Biol Chem 272:7211–7222PubMedCrossRefGoogle Scholar
  18. 18.
    Liu XH, Kirschenbaum A, Lu M, Yao S, Dosoretz A, Holland JF, Levine AC (2002) Prostaglandin E2 induces hypoxia-inducible factor-1alpha stabilization and nuclear localization in a human prostate cancer cell line. J Biol Chem 277:50081–50086PubMedCrossRefGoogle Scholar
  19. 19.
    Luetterforst R, Stang E, Zorzi N, Carozzi A, Way M, Parton RG (1999) Molecular characterization of caveolin association with the Golgi complex: identification of a cis-Golgi targeting domain in the caveolin molecule. J Cell Biol 145:1443–1459PubMedCrossRefGoogle Scholar
  20. 20.
    Murakami M, Kambe T, Shimbara S, Kudo I (1999) Functional coupling between various phospholipase A2s and cyclooxygenases in immediate and delayed prostanoid biosynthetic pathways. J Biol Chem 274:3103–3115PubMedCrossRefGoogle Scholar
  21. 21.
    Oh P, McIntosh DP, Schnitzer JE (1998) Dynamin at the neck of caveolae mediates their budding to form transport vesicles by GTP-driven fission from the plasma membrane of endothelium. J Cell Biol 141:101–114PubMedCrossRefGoogle Scholar
  22. 22.
    Okamoto T, Schlegel A, Scherer PE, Lisanti MP (1998) Caveolins, a family of scaffolding proteins for organizing “preassembled signaling complexes” at the plasma membrane. J Biol Chem 273:5419–5422PubMedCrossRefGoogle Scholar
  23. 23.
    Park SS, Kim JE, Kim YA, Kim YC, Kim SW (2005) Caveolin-1 is down-regulated and inversely correlated with HER2 and EGFR expression status in invasive ductal carcinoma of the breast. Histopathology 47:625–630PubMedCrossRefGoogle Scholar
  24. 24.
    Perrone G, Santini D, Vincenzi B, Zagami M, La Cesa A, Bianchi A, Altomare V, Primavera A, Battista C, Vetrani A, Tonini G, Rabitti C (2005) COX-2 expression in DCIS: correlation with VEGF, HER-2/neu, prognostic molecular markers and clinicopathological features. Histopathology 46:561–568PubMedCrossRefGoogle Scholar
  25. 25.
    Perrone G, Zagami M, Santini D, Vincenzi B, Gullotta G, Morini S, Battista C, Guerriero G, Altomare V, Tonini G, Rabitti C (2007) Cox-2 expression in lobular in situ neoplasia of the breast: correlation with histopathological grading system according to Tavassoli classification. Histopathology 51:33–39PubMedCrossRefGoogle Scholar
  26. 26.
    Picot D, Loll PJ, Garavito RM (1994) The X-ray crystal structure of the membrane protein prostaglandin H2 synthase-1. Nature 367:243–249PubMedCrossRefGoogle Scholar
  27. 27.
    Razandi M, Oh P, Pedram A, Schnitzer J, Levin ER (2002) ERs associate with and regulate the production of caveolin: implications for signaling and cellular actions. Mol Endocrinol 16:100–115PubMedCrossRefGoogle Scholar
  28. 28.
    Razani B, Engelman JA, Wang XB, Schubert W, Zhang XL, Marks CB, Macaluso F, Russell RG, Li M, Pestell RG, Di Vizio D, Hou H Jr, Kneitz B, Lagaud G, Christ GJ, Edelmann W, Lisanti MP (2001) Caveolin-1 null mice are viable but show evidence of hyperproliferative and vascular abnormalities. J Biol Chem 276:38121–38138PubMedCrossRefGoogle Scholar
  29. 29.
    Sagara Y, Mimori K, Yoshinaga K, Tanaka F, Nishida K, Ohno S, Inoue H, Mori M (2004) Clinical significance of Caveolin-1, Caveolin-2 and HER2/neu mRNA expression in human breast cancer. Br J Cancer 91:959–965PubMedGoogle Scholar
  30. 30.
    Spencer AG, Woods JW, Arakawa T, Singer II, Smith WL (1998) Subcellular localization of prostaglandin endoperoxide H synthases-1 and -2 by immunoelectron microscopy. J Biol Chem 273:9886–9893PubMedCrossRefGoogle Scholar
  31. 31.
    Song KS, Li S, Okamoto T, Quilliam LA, Sargiacomo M, Lisanti MP (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 Che 271:9690–9697CrossRefGoogle Scholar
  32. 32.
    Tavassoli FA, Millis RR, Boecker W, Lakhani SR (2003) Lobular neoplasia. In: Tavassoli FA, Devilee P (eds) World Health Organization classification of tumours. Pathology and genetics of tumours of the breast and female genital organs. IARC Press, Lyon, pp 60–62Google Scholar
  33. 33.
    Tsujii M, DuBois RN (1995) Alterations in cellular adhesion and apoptosis in epithelial cells overexpressing prostaglandin endoperoxide synthase 2. Cell 83:493–501PubMedCrossRefGoogle Scholar
  34. 34.
    Williams TM, Sotgia F, Lee H, Hassan G, Di Vizio D, Bonuccelli G, Capozza F, Mercier I, Rui H, Pestell RG, Lisanti MP (2006) Stromal and epithelial caveolin-1 both confer a protective effect against mammary hyperplasia and tumorigenesis: caveolin-1 antagonizes cyclin D1 function in mammary epithelial cells. Am J Pathol 169:1784–1801PubMedCrossRefGoogle Scholar
  35. 35.
    Zhao Y, Agarwal VR, Mendelson CR, Simpson ER (1996) Estrogen biosynthesis proximal to a breast tumor is stimulated by PGE2 via cyclic AMP, leading to activation of promoter II of the CYP19 (aromatase) gene. Endocrinology 137:5739–5742PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Giuseppe Perrone
    • 1
  • Mariagiovanna Zagami
    • 1
  • Vittorio Altomare
    • 2
  • Cleonice Battista
    • 2
  • Sergio Morini
    • 3
  • Carla Rabitti
    • 1
  1. 1.Surgical PathologyUniversità Campus Bio-Medico di RomaRomeItaly
  2. 2.Breast Surgical UnitUniversità Campus Bio-Medico di RomaRomeItaly
  3. 3.Department of Biomedical ResearchUniversità Campus Bio-Medico di RomaRomeItaly

Personalised recommendations