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Caveolin-1 Expression Together with VEGF can be a Predictor for Lung Metastasis and Poor Prognosis in Osteosarcoma

  • Fatma El-Zahraa Ammar Mohamed
  • El Zahraa Ibrahim KhalilEmail author
  • Nisreen D. M. Toni
Original Article
  • 41 Downloads

Abstract

Caveolin-1, the major protein component of caveolae, plays vital functions in tumorigenesis and metastasis. Previous evidence demonstrated the positive role of Caveolin-1 in the regulation of endothelial cell differentiation and the involvement of Caveolin-1 in vascular endothelial growth factor (VEGF) mediated angiogenesis. The correlation of Caveolin-1 expression and angiogenesis is not yet elucidated in osteosarcoma. This study aimed to investigate the expression levels of Caveolin-1 and VEGF in osteosarcoma and their associations with clinicopathological data. This study included 66 formalin-fixed and paraffin embedded osteosarcoma tissue samples. The expression levels of Caveolin-1 and VEGF were assessed by immunohistochemistry. Then associations with clinicopathological variables and the correlation between both markers were evaluated statistically. We also investigated the expression of Caveolin-1 and VEGF values in gene microarrays of osteosarcoma patients and cell lines by using GEO data sets on https://www.ncbi.nlm.nih.gov. Caveolin-1 and VEGF were expressed in 19.6% and 77.3%, respectively. Caveolin-1 expression was associated positively with osteoblastic histological subtype (P < 0.0001). VEGF expression showed positive association with patient age, histological grade and clinical stage (P = 0.031, P = 0.024 and P < 0.001; respectively). An inverse correlation between Caveolin-1 and VEGF expressions in osteosarcoma was found (r = 0.2 P = 0.04). In silico analysis of Caveolin-1 and VEGF expression supported our results. Our results suggest that Caveolin-1 may act as a tumor suppressor in osteosarcoma. Down-regulation of Caveolin-1 can be used as an indicator for poor prognosis in osteosarcoma patients. Meanwhile, overexpression of VEGF is a predictor of pulmonary metastasis and poor prognosis.

Keywords

Caveolin-1 Vascular endothelial growth factor Osteosarcoma Immunohistochemistry 

Notes

Author contributions

All authors contribute equally in this work.

Funding

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Compliance with ethical standards

Conflict of Interests

All the authors have no potential conflicts (financial, professional, or personal) relevant to the manuscript to disclose.

References

  1. 1.
    Isakoff MS, Bielack SS, Meltzer P, Gorlick R (2015) Osteosarcoma: Current treatment and a collaborative pathway to success. J Clin Oncol 33:3029–3035.  https://doi.org/10.1200/JCO.2014.59.4895 CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Klein MJ, Siegal GP (2006) Osteosarcoma: anatomic and histologic variants. Am J Clin Pathol 125(4):555–581.  https://doi.org/10.1309/UC6K-QHLD-9LV2-KENN CrossRefPubMedGoogle Scholar
  3. 3.
    Rubin P, Williams JP, Devesa SS, Travis LB, Constine LS (2010) Cancer genesis across the age spectrum: associations with tissue development, maintenance, and senescence. Semin Radiat Oncol 20:3–11.  https://doi.org/10.1016/j.semradonc.2009.08.001 CrossRefPubMedGoogle Scholar
  4. 4.
    Fletcher CD, Bridge J, Hogendoorn PC, Mertens F (eds) (2013) The World Health Organization Classification of Tumors of Soft Tissue and Bone. IARC, LyonGoogle Scholar
  5. 5.
    Zalupski MM, Rankin C, Ryan JR, Lucas DR, Muler J, Lanier KS, Budd GT, Biermann JS, Meyers FJ, Antman K (2004) Adjuvant therapy of osteosarcoma–a phase II trial: Southwest Oncology Group study 9139. Cancer 100(4):818–825.  https://doi.org/10.1002/cncr.20021 CrossRefPubMedGoogle Scholar
  6. 6.
    Wada T, Isu K, Takeda N, Usui M, Ishii S, Yamawaki S (1996) A preliminary report of neoadjuvant chemotherapy NSH-7 study in osteosarcoma: preoperative salvage chemotherapy based on clinical tumor response and the use of granulocyte colony-stimulating factor. Oncology 53:221–227.  https://doi.org/10.1159/000227564 CrossRefPubMedGoogle Scholar
  7. 7.
    Xie Y, Huang J, Wu M, Zhou Y (2018) Expression of CD133 protein in osteosarcoma and its relationship with the clinicopathological features and prognosis. J Cancer Res Ther 14:892–895.  https://doi.org/10.4103/jcrt.JCRT_461_17 CrossRefPubMedGoogle Scholar
  8. 8.
    Xu M, Xie Y, Sheng W, Miao J, Yang J (2015) Adenovirus-mediated ING4 Gene Transfer in Osteosarcoma Suppresses Tumor Growth via Induction of Apoptosis and Inhibition of Tumor Angiogenesis. Technology in Cancer Research and Treatment 14:369–378.  https://doi.org/10.1177/1533034614500424 CrossRefPubMedGoogle Scholar
  9. 9.
    Kaya M, Wada T, Akatsuka T, Kawaguchi S, Nagoya S, Shindoh M, Higashino F, Mezawa F, Okada F, Ishii S (2000) Vascular endothelial growth factor expression in untreated osteosarcoma is predictive of pulmonary metastasis and poor prognosis. Clin Cancer Res 6(2):572–577PubMedGoogle Scholar
  10. 10.
    DuBois S, Demetri G (2007) Markers of angiogenesis and clinical features in patientswith sarcoma. Cancer 109(5):813–981.  https://doi.org/10.1002/cncr.22455 CrossRefPubMedGoogle Scholar
  11. 11.
    Dvorak HF, Sioussat TM, Brown LF, Berse B, Nagy JA, Sotrel A, Manseau EJ, Van de Water L, Senger DR (1991) Distribution of vascular permeability factor (vascular endothelial growth factor) in tumors: concentration in tumor blood vessels. J Exp Med 174(5):1275–1278CrossRefGoogle Scholar
  12. 12.
    Cohen AW, Hnasko R, Schubert W, Lisanti MP (2004) Role of caveolae and caveolins in health and disease. Physiol Rev 84:1341–1379.  https://doi.org/10.1152/physrev.00046.2003 CrossRefGoogle Scholar
  13. 13.
    Schwencke C, Braun-Dullaeus RC, Wunderlich C, Strasser RH (2006) Caveolae and caveolin in transmembrane signaling: Implications for human disease. Cardiovasc Res 70(1):42–49.  https://doi.org/10.1016/j.cardiores.2005.11.029 CrossRefPubMedGoogle Scholar
  14. 14.
    Stan RV (2005) Structure of caveolae. Journal of Biochimica et Biophysica Acta 1746:334–348.  https://doi.org/10.1016/j.bbamcr.2005.08.008 CrossRefPubMedGoogle Scholar
  15. 15.
    Smart EJ, Graf GA, McNiven MA, Sessa WC, Engelman JA, Scherer PE, Okamoto T, Lisanti MP (1999) Caveolins, liquid-ordered domains, and signal transduction. Mol Cell Biol 19:7289–7304CrossRefGoogle Scholar
  16. 16.
    Solomon KR, Danciu TE, Adolphson LD, Hecht LE, Hauschka PV (2000) Caveolin-enriched membrane signaling complexes in human and murine osteoblasts. J Bone Miner Res 15:2380–2390.  https://doi.org/10.1359/jbmr.2000.15.12.2380 CrossRefPubMedGoogle Scholar
  17. 17.
    Solomon KR, Adolphson LD, Wank DA, McHugh KP, Hauschka PV (2000) Caveolae in human and murine osteoblasts. J Bone Miner Res 15:2391–2401.  https://doi.org/10.1359/jbmr.2000.15.12.2391 CrossRefPubMedGoogle Scholar
  18. 18.
    Wiechen K, Sers C, Agoulnik A, Arlt K, Dietel M, Schlag PM, Schneider U (2001) Down-regulation of caveolin-1, a candidate tumor suppressor gene, in sarcomas. Am J Pathol 158:833–839.  https://doi.org/10.1016/S0002-9440(10)64031-X CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Belanger MM, Roussel E, Couet J (2004) Caveolin-1 is down-regulated in human lung carcinoma and acts as a candidate tumor suppressor gene. Chest 125(5 suppl):106S.  https://doi.org/10.1378/chest.125.5_suppl.106S CrossRefPubMedGoogle Scholar
  20. 20.
    Wiechen K, Diatchenko L, Agoulnik A, Scharff KM, Schober H, Arlt K, Zhumabayeva B, Siebert PD, Dietel M, Schäfer R, Sers C (2001) Caveolin-1 is down-regulated in human ovarian carcinoma and acts as a candidate tumor suppressor gene. Am J Pathol 159:1635–1643.  https://doi.org/10.1016/S0002-9440(10)63010-6 CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Kato K, Hida Y, Miyamoto M, Hashida H, Shinohara T, Itoh T, Okushiba S, Kondo S, Katoh H (2002) Overexpression of caveolin-1 in esophageal squamous cell carcinoma correlates with lymph node metastasis and pathologic stage. Cancer 94:929–933.  https://doi.org/10.1002/cncr.10329 CrossRefPubMedGoogle Scholar
  22. 22.
    Yang G, Truong LD, Wheeler TM, Thompson TC (1999) Caveolin- 1 expression in clinically confined human prostate cancer: a novel prognostic marker. Cancer Res 59:5719–5723PubMedGoogle Scholar
  23. 23.
    Cantiani L, Manara MC, Zucchini C, De Sanctis P, Zuntini M, Valvassori L, Serra M, Olivero M, Di Renzo MF, Colombo MP, Picci P, Scotlandi K (2007) Caveolin-1 Reduces Osteosarcoma Metastases by Inhibiting c-Src Activity and Met Signaling. Cancer Res 67(16):7675–7685.  https://doi.org/10.1158/0008-5472.CAN-06-4697 CrossRefPubMedGoogle Scholar
  24. 24.
    Carver LA, Schnitzer JE (2003) Caveolae: mining little caves for new cancer targets. Nat Rev Cancer 3:571–581.  https://doi.org/10.1038/nrc1146 CrossRefPubMedGoogle Scholar
  25. 25.
    Krajewska WM, Maslowska I (2004) Caveolins: structure and function in signal transduction. Cell Mol Biol Lett 9:195–220Google Scholar
  26. 26.
    Williams TM, Lisanti MP (2005) Caveolin-1 in oncogenic transformation, cancer, and metastasis. Am J Physiol Cell Physiol 288:494–506.  https://doi.org/10.1152/ajpcell.00458.2004 CrossRefGoogle Scholar
  27. 27.
    Liu J, Wang XB, Park DS, Lisanti MP (2002) Caveolin-1 expression enhances endothelial capillary tubule formation. J Biol Chem 277:10661–10668.  https://doi.org/10.1074/jbc.M110354200 CrossRefPubMedGoogle Scholar
  28. 28.
    Deurs B, Roepstorff K, Hommelgaard AM, Sandvig K (2003) Caveolae: anchored, multifunctional platforms in the lipid ocean. Trends Cell Biol 13:92–100.  https://doi.org/10.1016/S0962-8924(02)00039-9 CrossRefPubMedGoogle Scholar
  29. 29.
    Razani B, Woodman SE, Lisanti MP (2002) Caveolae: from cell biology to animal physiology. Pharmacol Rev 54:431–467CrossRefGoogle Scholar
  30. 30.
    Feron O, Kelly RA (2001) The caveolar paradox: suppressing, inducing, and terminating eNOS signaling. Circ Res 88:129–131CrossRefGoogle Scholar
  31. 31.
    Chen J, Braet F, Brodsky S, Weinstein T, Romanov V, Noiri E, Goligorsky MS (2002) VEGF-induced mobilization of caveolae and increase in permeability of endothelial cells. Am J Phys 282(5):C1053–C1063.  https://doi.org/10.1152/ajpcell.00292.2001 CrossRefGoogle Scholar
  32. 32.
    Shi L, Chen XM, Wang L, Zhang L, Chen Z (2007) Expression of Caveolin-1 in Mucoepidermoid Carcinoma of the Salivary Glands: Correlation with Vascular Endothelial Growth Factor, Microvessel Density, and Clinical Outcome. CANCER 109(8):1523–1531.  https://doi.org/10.1002/cncr.22573 CrossRefPubMedGoogle Scholar
  33. 33.
    Tahir SA, Park S, Thompson TC (2009) Caveolin-1 regulates VEGF-stimulated angiogenic activities in prostate cancer and endothelial cells. Cancer Biol Ther 8(23):2286–2296CrossRefGoogle Scholar
  34. 34.
    Gamallo C, Palacios J, Moreno G, Calvo de Mora J, Suarez A, Armas A (1999) Beta-catenin expression pattern in stage I and II ovarian carcinomas: relationship with beta-catenin gene mutations, clinicopathological features, and clinical outcome. Am J Pathol 155:527–536CrossRefGoogle Scholar
  35. 35.
    Galbiati F, Volonte D, Engelman JA et al (1998) Targeted downregulation of caveolin-1 is sufficient to drive cell transformation and hyperactivate the p42/44 MAP kinase cascade. EMBO J 17:6633–6648CrossRefGoogle Scholar
  36. 36.
    Koleske AJ, Baltimore D, Lisanti MP (1995) Reduction of caveolin and caveolae in oncogenically transformed cells. Proc Natl Acad Sci U S A 92:1381–1385CrossRefGoogle Scholar
  37. 37.
    Sloan EK, Stanley KL, Anderson RL (2004) Caveolin-1 inhibits breast cancer growth and metastasis. Oncogene 23:7893–7897CrossRefGoogle Scholar
  38. 38.
    Hu YC, Lam KY, Law S et al (2001) Profiling of differentially expressed cancer-related genes in esophageal squamous cell carcinoma (ESCC) using human cancer cDNA arrays: overexpression of oncogene MET correlates with tumor differentiation in ESCC. Clin Cancer Res 7:3519–3525PubMedGoogle Scholar
  39. 39.
    Fong A, Garcia E, Gwynn L et al (2003) Expression of caveolin-1 and caveolin-2 in urothelial carcinoma of the urinary bladder correlates with tumor grade and squamous differentiation. Am J Clin Pathol 120:93–100CrossRefGoogle Scholar
  40. 40.
    Li L, Yang G, Ebara S et al (2001) Caveolin-1 mediates testosterone stimulate survival/clonal growth and promotes metastatic activities in prostate cancer cells. Cancer Res 61:4386–4392PubMedGoogle Scholar
  41. 41.
    Tirado OM, Mateo-Lozano S, Villar J, Dettin LE, Llort A, Gallego S, Ban J, Kovar H, Notario V (2006) Caveolin-1 (CAV1) is a target of EWS/FLI-1 and a key determinant of the oncogenic phenotype and tumorigenicity of Ewing’s sarcoma cells. Cancer Res 66:9937–9947.  https://doi.org/10.1158/0008-5472.CAN-06-0927 CrossRefPubMedGoogle Scholar
  42. 42.
    Lee H, Volonte D, Galbiati F, Lyengar P, Lublin MD, Bregman DB, Wilson MT, Campos-Gonzalez R, Bouzahzah B, Pestell RG, Scherer PE, Lisanti MP (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 14:1750–1775.  https://doi.org/10.1210/mend.14.11.0553 CrossRefPubMedGoogle Scholar
  43. 43.
    Oda Y, Yamamoto H, Tamiya S, Matsuda S, Tanaka K, Yokoyama R, Iwamoto Y, Tsuneyoshi M (2006) CXCR4 and VEGF expression in the primary site and the metastatic site of human osteosarcoma: analysis within a group of patients, all of whom developed lung metastasis. Mod Pathol 19(5):738–745.  https://doi.org/10.1038/modpathol.3800587 CrossRefPubMedGoogle Scholar
  44. 44.
    Bajpai J, Sharma M, Sreenivas V, Kumar R, Gamnagatti S, Khan SA, Rastogi S, Malhotra A, Bakhshi S (2009) VEGF expression as a prognostic marker in osteosarcoma. Pediatr Blood Cancer 53(6):1035–1039.  https://doi.org/10.1002/pbc.22178 CrossRefPubMedGoogle Scholar
  45. 45.
    Lammli J, Fan M, Rosenthal HG, Patni M, Rinehart E, Vergara G, Ablah E, Wooley PH, Lucas G, Yang SY (2012) Expression of Vascular Endothelial Growth Factor correlates with the advance of clinical osteosarcoma. Int Orthop 36(11):2307–2313.  https://doi.org/10.1007/s00264-012-1629-z CrossRefPubMedPubMedCentralGoogle Scholar
  46. 46.
    Ługowska I, Woźniak W, Klepacka T, Michalak E, Szamotulska K (2011) A prognostic evaluation of vascular endothelial growth factor in children and young adults with osteosarcoma. Pediatr Blood Cancer 57(1):63–68.  https://doi.org/10.1002/pbc.23021 CrossRefPubMedGoogle Scholar
  47. 47.
    Becker RG, Galia CR, Morini S, Viana CR (2013) Immunohistochemical expression of VEGF and her-2 proteins in osteosarcoma biopsies. Acta Ortopedica Brasileira 21(4):233–238.  https://doi.org/10.1590/S1413-78522013000400010 CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Baptista AM, Camargo AF, Filippi RZ, Oliveira CR, Azevedo Neto RS, Camargo OP (2014) Correlation between the expression of vegf and Survival in osteosarcoma. Acta Ortopedica Brasileira 22(5):250–255.  https://doi.org/10.1590/1413-78522014220500978 CrossRefGoogle Scholar
  49. 49.
    Zhao J, Zhang Z, Zhao N, Ma BA, Fan QU (2015) VEGF silencing inhibits human osteosarcoma angiogenesis and promotes cell apoptosis via PI3K/AKT signaling pathway. Cell Biochem Biophys 73(2):519–525.  https://doi.org/10.1007/s12013-015-0692-7 CrossRefPubMedGoogle Scholar
  50. 50.
    Mizobuchi H, García-Castellano JM, Philip S, Healey JH, Gorlick R (2008) Hypoxia markers in human osteosarcoma: an exploratory study. Clin Orthop Relat Res 466(9):2052–2059.  https://doi.org/10.1007/s11999-008-0328-y CrossRefPubMedPubMedCentralGoogle Scholar
  51. 51.
    Zhou Q, Zhu Y, Deng Z, Long H, Zhang S, Chen X (2011) VEGF and EMMPRIN expression correlates with survival of patients with osteosarcoma. Surg Oncol 20(1):13–19.  https://doi.org/10.1016/j.suronc.2009.09.002 CrossRefPubMedGoogle Scholar
  52. 52.
    Hassan SE, Bekarev M, Kim MY, Lin J, Piperdi S, Gorlick R, Geller DS (2012) Cell surface receptor expression patterns in osteosarcoma. Cancer 118:740–749.  https://doi.org/10.1002/cncr.26339 CrossRefPubMedGoogle Scholar
  53. 53.
    Chen D, Zhang YJ, Zhu KW, Wang WC (2013) A systematic review of vascular endothelial growth factor expression as a biomarker of prognosis in patients with osteosarcoma. Tumour Biol 34(3):1895–1899.  https://doi.org/10.1007/s13277-013-0733-z CrossRefPubMedGoogle Scholar
  54. 54.
    Ohba T, Cates JM, Cole HA, Slosky DA, Haro H, Ando T, Schwartz HS, Schoenecker JG (2014) Autocrine VEGF/VEGFR1 signaling in a subpopulation of cells associates with aggressive osteosarcoma. Mol Cancer Res 12:1100–1111.  https://doi.org/10.1158/1541-7786.MCR-14-0037 CrossRefPubMedGoogle Scholar
  55. 55.
    Yu XW, Wu TY, Yi X, Ren WP, Zhou ZB, Sun YQ, Zhang CQ (2014) Prognostic significance of VEGF expression in osteosarcoma: a meta-analysis. Tumour Biol 35:155–160.  https://doi.org/10.1007/s13277-013-1019-1 CrossRefPubMedGoogle Scholar
  56. 56.
    Daft PG, Yang Y, Napierala D, Zayzafoon M (2015) The Growth and Aggressive Behavior of Human Osteosarcoma Is Regulated by a CaMKII-Controlled Autocrine VEGF Signaling Mechanism. PLoS One 10(4):1–20. e0121568.  https://doi.org/10.1371/journal.Pone CrossRefGoogle Scholar
  57. 57.
    Tang Y, Zeng X, He F, Liao Y, Qian N, Toi M (2012) Caveolin-1 is related to invasion, survival, and poor prognosis in hepatocellular cancer. Med Oncol 29(2):977–984.  https://doi.org/10.1007/s12032-011-9900-5 CrossRefPubMedGoogle Scholar
  58. 58.
    Hoffman R (2004) Do the signalling proteins for angiogenesis exist as a modular complex? The case for the angosome. Med Hypotheses 63:675–680CrossRefGoogle Scholar
  59. 59.
    Wu T, Zhang B, Ye F, Xiao Z (2013) A potential role for caveolin-1 in VEGF-induced fibronectin upregulation in mesangial cells: involvement of VEGFR2 and Src. Am J Physiol Ren Physiol 304:F820–F830.  https://doi.org/10.1152/ajprenal.00294.2012 CrossRefGoogle Scholar
  60. 60.
    Liu J, Razani B, Tang S, Terman BI, Ware JA, Lisanti MP (1999) Angiogenesis activators and inhibitors differentially regulate caveolin-1 expression and caveolae formation in vascular endothelial cells. Angiogenesis inhibitors block vascular endothelial growth factor-induced downregulation of caveolin-1. J Biol Chem 274:15781–15785.  https://doi.org/10.1074/jbc.274.22.15781 CrossRefPubMedGoogle Scholar

Copyright information

© Arányi Lajos Foundation 2019

Authors and Affiliations

  • Fatma El-Zahraa Ammar Mohamed
    • 1
  • El Zahraa Ibrahim Khalil
    • 1
    Email author
  • Nisreen D. M. Toni
    • 1
  1. 1.Pathology Department, Faculty of MedicineMinia UniversityMiniaEgypt

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