Tumor Biology

, Volume 32, Issue 1, pp 23–32 | Cite as

Clinical significance of Maspin promoter methylation and loss of its protein expression in invasive ductal breast carcinoma: correlation with VEGF-A and MTA1 expression

  • Gayatri Sharma
  • Sameer Mirza
  • Rajinder Parshad
  • Anurag Srivastava
  • Siddartha Datta Gupta
  • Pranav Pandya
  • Ranju Ralhan
Research Article

Abstract

Maspin is a serine protease inhibitor with tumor-suppressor activity. Maspin can suppress tumor growth and metastasis in vivo and tumor cell motility and invasion in vitro. Previous studies indicate that the loss of Maspin expression is closely linked to aberrant methylation of the Maspin promoter. We examined the promoter methylation status of Maspin in tumor and corresponding serum of breast cancer patients. In addition, protein expression of this gene was also assessed to determine possible correlation between promoter hypermethylation and gene silencing. Further, we investigated the correlation of Maspin expression with vascular endothelial growth factor (VEGF-A) and MTA1 expression. Maspin methylation was analyzed by methylation-specific PCR in 100 invasive ductal breast carcinoma patients’ tumors and circulating DNA in a prospective study. Promoter hypermethylation was correlated with expression of the encoded protein in tumors by immunohistochemistry. Significant correlation was observed between promoter hypermethylation of Maspin (r = +0.88; p ≤ 0.0001) in tumors and paired sera. Significant association was found between Maspin promoter hypermethylation and loss of its protein expression (p = 0.01, OR = 3.1, 95% CI = 1.3–7.4). The expression of VEGF-A and MTA1 was lower in tumors with high Maspin expression compared to tumors with loss of Maspin expression. Our results indicate that aberrant promoter methylation is associated with loss of Maspin immunoreactivity in breast cancer tissues. Further, loss of Maspin expression is significantly correlated with increased expression of VEGF-A and MTA1.

Keywords

Breast cancer DNA methylation Maspin MTA1 Gene silencing 

References

  1. 1.
    Sheng S, Carey J, Seftor EA, Dias L, Hendrix MJ, Sager R. Maspin acts at the cell membrane to inhibit invasion and motility of mammary and prostatic cancer cells. Proc Natl Acad Sci USA. 1996;93:11669–74.CrossRefPubMedGoogle Scholar
  2. 2.
    Liu J, Yin S, Reddy N, Spencer C, Sheng S. Bax mediates the apoptosis-sensitizing effect of maspin. Cancer Res. 2004;64:1703–11.CrossRefPubMedGoogle Scholar
  3. 3.
    Jiang N, Meng Y, Zhang S, Mensah-Osman E, Sheng S. Maspin sensitizes breast carcinoma cells to induced apoptosis. Oncogene. 2002;21:4089–98.CrossRefPubMedGoogle Scholar
  4. 4.
    Zhang M, Volpert O, Shi YH, Bouck N. Maspin is an angiogenesis inhibitor. Nat Med. 2000;6:196–9.CrossRefPubMedGoogle Scholar
  5. 5.
    Domann FE, Rice JC, Hendrix MJ, Futscher BW. Epigenetic silencing of maspin gene expression in human breast cancers. Int J Cancer. 2000;85:805–10.CrossRefPubMedGoogle Scholar
  6. 6.
    Maass N, Biallek M, Rosel F, Schem C, Ohike N, Zhang M, et al. Hypermethylation and histone deacetylation lead to silencing of the maspin gene in human breast cancer. Biochem Biophys Res Commun. 2002;297:125–8.CrossRefPubMedGoogle Scholar
  7. 7.
    Primeau M, Gagnon J, Momparler RL. Synergistic antineoplastic action of DNA methylation inhibitor 5-aza-2'-deoxycytidine and histone deacetylase inhibitor depsipeptide on human breast carcinoma cells. Int J Cancer. 2003;103:177–84.CrossRefPubMedGoogle Scholar
  8. 8.
    Khalkhali-Ellis Z. Maspin: the new frontier. Clin Cancer Res. 2006;12:7279–83.CrossRefPubMedGoogle Scholar
  9. 9.
    Akiyama Y, Maesawa C, Ogasawara S, Terashima M, Masuda T. Cell-type-specific repression of the maspin gene is disrupted frequently by demethylation at the promoter region in gastric intestinal metaplasia and cancer cells. Am J Pathol. 2003;163:1911–9.PubMedGoogle Scholar
  10. 10.
    Bettstetter M, Woenckhaus M, Wild PJ, Rummele P, Blaszyk H, Hartmann A, et al. Elevated nuclear maspin expression is associated with microsatellite instability and high tumour grade in colorectal cancer. J Pathol. 2005;205:606–14.CrossRefPubMedGoogle Scholar
  11. 11.
    Boltze C, Schneider-Stock R, Quednow C, Hinze R, Mawrin C, Hribaschek A, et al. Silencing of the maspin gene by promoter hypermethylation in thyroid cancer. Int J Mol Med. 2003;12:479–84.PubMedGoogle Scholar
  12. 12.
    Goelz SE, Vogelstein B, Hamilton SR, Feinberg AP. Hypomethylation of DNA from benign and malignant human colon neoplasms. Science. 1985;228:187–90.CrossRefPubMedGoogle Scholar
  13. 13.
    Reis-Filho JS, Torio B, Albergaria A, Schmitt FC. Maspin expression in normal skin and usual cutaneous carcinomas. Virchows Arch. 2002;441:551–8.CrossRefPubMedGoogle Scholar
  14. 14.
    Rose SL, Fitzgerald MP, White NO, Hitchler MJ, Futscher BW, De Geest K, et al. Epigenetic regulation of maspin expression in human ovarian carcinoma cells. Gynecol Oncol. 2006;102:319–24.CrossRefPubMedGoogle Scholar
  15. 15.
    Sato N, Maitra A, Fukushima N, van Heek NT, Matsubayashi H, Iacobuzio-Donahue CA, et al. Frequent hypomethylation of multiple genes overexpressed in pancreatic ductal adenocarcinoma. Cancer Res. 2003;63:4158–66.PubMedGoogle Scholar
  16. 16.
    Bolat F, Gumurdulu D, Erkanli S, Kayaselcuk F, Zeren H, Ali Vardar M, et al. Maspin overexpression correlates with increased expression of vascular endothelial growth factors a, c, and d in human ovarian carcinoma. Pathol Res Pract. 2008;204:379–87.CrossRefPubMedGoogle Scholar
  17. 17.
    Chua R, Setzer S, Govindarajan B, Sexton D, Cohen C, Arbiser JL. Maspin expression, angiogenesis, prognostic parameters, and outcome in malignant melanoma. J Am Acad Dermatol. 2009;60:758–66.CrossRefPubMedGoogle Scholar
  18. 18.
    Zhang H, Stephens LC, Kumar R. Metastasis tumor antigen family proteins during breast cancer progression and metastasis in a reliable mouse model for human breast cancer. Clin Cancer Res. 2006;12:1479–86.CrossRefPubMedGoogle Scholar
  19. 19.
    Balasenthil S, Broaddus RR, Kumar R. Expression of metastasis-associated protein 1 (mta1) in benign endometrium and endometrial adenocarcinomas. Hum Pathol. 2006;37:656–61.CrossRefPubMedGoogle Scholar
  20. 20.
    Hofer MD, Kuefer R, Varambally S, Li H, Ma J, Shapiro GI, et al. The role of metastasis-associated protein 1 in prostate cancer progression. Cancer Res. 2004;64:825–9.CrossRefPubMedGoogle Scholar
  21. 21.
    Kumar R, Wang RA, Mazumdar A, Talukder AH, Mandal M, Yang Z, et al. A naturally occurring mta1 variant sequesters oestrogen receptor-alpha in the cytoplasm. Nature. 2002;418:654–7.CrossRefPubMedGoogle Scholar
  22. 22.
    Mazumdar A, Wang RA, Mishra SK, Adam L, Bagheri-Yarmand R, Mandal M, et al. Transcriptional repression of oestrogen receptor by metastasis-associated protein 1 corepressor. Nat Cell Biol. 2001;3:30–7.CrossRefPubMedGoogle Scholar
  23. 23.
    Nicolson GL, Nawa A, Toh Y, Taniguchi S, Nishimori K, Moustafa A. Tumor metastasis-associated human mta1 gene and its mta1 protein product: role in epithelial cancer cell invasion, proliferation and nuclear regulation. Clin Exp Metastasis. 2003;20:19–24.CrossRefPubMedGoogle Scholar
  24. 24.
    Lewis CM, Cler LR, Bu DW, Zochbauer-Muller S, Milchgrub S, Naftalis EZ, et al. Promoter hypermethylation in benign breast epithelium in relation to predicted breast cancer risk. Clin Cancer Res. 2005;11:166–72.PubMedGoogle Scholar
  25. 25.
    Frommer M, McDonald LE, Millar DS, Collis CM, Watt F, Grigg GW, et al. A genomic sequencing protocol that yields a positive display of 5-methylcytosine residues in individual DNA strands. Proc Natl Acad Sci USA. 1992;89:1827–31.CrossRefPubMedGoogle Scholar
  26. 26.
    Murai S, Maesawa C, Masuda T, Sugiyama T. Aberrant maspin expression in human endometrial cancer. Cancer Sci. 2006;97(9):883–8.CrossRefPubMedGoogle Scholar
  27. 27.
    Sharma G, Mirza S, Prasad CP, Srivastava A, Gupta SD, Ralhan R. Promoter hypermethylation of p16ink4a, p14arf, cyclind2 and slit2 in serum and tumor DNA from breast cancer patients. Life Sci. 2007;80:1873–81.CrossRefPubMedGoogle Scholar
  28. 28.
    Mohsin SK, Zhang M, Clark GM, Craig Allred D. Maspin expression in invasive breast cancer: association with other prognostic factors. J Pathol. 2003;199:432–5.CrossRefPubMedGoogle Scholar
  29. 29.
    Jang KS, Paik SS, Chung H, Oh YH, Kong G. Mta1 overexpression correlates significantly with tumor grade and angiogenesis in human breast cancers. Cancer Sci. 2006;97:374–9.CrossRefPubMedGoogle Scholar
  30. 30.
    Goldhirsch A, Wood WC, Gelber RD, Coates AS, Thurlimann B, Senn HJ. Meeting highlights: updated international expert consensus on the primary therapy of early breast cancer. J Clin Oncol. 2003;21:3357–65.CrossRefPubMedGoogle Scholar
  31. 31.
    Ryden L, Stendahl M, Jonsson H, Emdin S, Bengtsson NO, Landberg G. Tumor-specific vegf-a and vegfr2 in postmenopausal breast cancer patients with long-term follow-up. Implication of a link between vegf pathway and tamoxifen response. Breast Cancer Res Treat. 2005;89:135–43.CrossRefPubMedGoogle Scholar
  32. 32.
    Rhodes A, Jasani B, Anderson E, Dodson AR, Balaton AJ. Evaluation of her-2/neu immunohistochemical assay sensitivity and scoring on formalin-fixed and paraffin-processed cell lines and breast tumors: a comparative study involving results from laboratories in 21 countries. Am J Clin Pathol. 2002;118:408–17.CrossRefPubMedGoogle Scholar
  33. 33.
    Fitzgerald M, Oshiro M, Holtan N, Krager K, Cullen JJ, Futscher BW, et al. Human pancreatic carcinoma cells activate maspin expression through loss of epigenetic control. Neoplasia. 2003;5:427–36.PubMedGoogle Scholar
  34. 34.
    Futscher BW, O'Meara MM, Kim CJ, Rennels MA, Lu D, Gruman LM, et al. Aberrant methylation of the maspin promoter is an early event in human breast cancer. Neoplasia. 2004;6:380–9.CrossRefPubMedGoogle Scholar
  35. 35.
    Khalkhali-Ellis Z, Hendrix MJ. Elucidating the function of secreted maspin: inhibiting cathepsin d-mediated matrix degradation. Cancer Res. 2007;67:3535–9.CrossRefPubMedGoogle Scholar
  36. 36.
    Cho JH, Kim HS, Park CS, Kim JK, Jung KY, Shin BK, et al. Maspin expression in early oral tongue cancer and its relation to expression of mutant-type p53 and vascular endothelial growth factor (vegf). Oral Oncol. 2007;43:272–7.CrossRefPubMedGoogle Scholar
  37. 37.
    Frey A, Soubani AO, Adam AK, Sheng S, Pass HI, Lonardo F. Nuclear, compared with combined nuclear and cytoplasmic expression of maspin, is linked in lung adenocarcinoma to reduced vegf-a levels and in stage i, improved survival. Histopathology. 2009;54:590–7.CrossRefPubMedGoogle Scholar
  38. 38.
    Solomon LA, Munkarah AR, Schimp VL, Arabi MH, Morris RT, Nassar H, et al. Maspin expression and localization impact on angiogenesis and prognosis in ovarian cancer. Gynecol Oncol. 2006;101:385–9.CrossRefPubMedGoogle Scholar
  39. 39.
    Song SY, Lee SK, Kim DH, Son HJ, Kim HJ, Lim YJ, et al. Expression of maspin in colon cancers: its relationship with p53 expression and microvessel density. Dig Dis Sci. 2002;47:1831–5.CrossRefPubMedGoogle Scholar
  40. 40.
    Terashima M, Maesawa C, Oyama K, Ohtani S, Akiyama Y, Ogasawara S, et al. Gene expression profiles in human gastric cancer: expression of maspin correlates with lymph node metastasis. Br J Cancer. 2005;92:1130–6.CrossRefPubMedGoogle Scholar
  41. 41.
    Reddy KB, McGowen R, Schuger L, Visscher D, Sheng S. Maspin expression inversely correlates with breast tumor progression in mmtv/tgf-alpha transgenic mouse model. Oncogene. 2001;20:6538–43.CrossRefPubMedGoogle Scholar
  42. 42.
    Shi HY, Zhang W, Liang R, Abraham S, Kittrell FS, Medina D, et al. Blocking tumor growth, invasion, and metastasis by maspin in a syngeneic breast cancer model. Cancer Res. 2001;61:6945–51.PubMedGoogle Scholar
  43. 43.
    Vecchi M, Confalonieri S, Nuciforo P, Vigano MA, Capra M, Bianchi M, et al. Breast cancer metastases are molecularly distinct from their primary tumors. Oncogene. 2008;27:2148–58.CrossRefPubMedGoogle Scholar
  44. 44.
    Sheng S. A role of novel serpin maspin in tumor progression: the divergence revealed through efforts to converge. J Cell Physiol. 2006;209:631–5.CrossRefPubMedGoogle Scholar
  45. 45.
    Zhang M. Toward re-expressing tumor suppressor gene maspin in breast cancer. Clin Breast Cancer. 2002;3:353–4.PubMedGoogle Scholar
  46. 46.
    Zou Z, Gao C, Nagaich AK, Connell T, Saito S, Moul JW, et al. P53 regulates the expression of the tumor suppressor gene maspin. J Biol Chem. 2000;275:6051–4.CrossRefPubMedGoogle Scholar
  47. 47.
    Maass N, Hojo T, Ueding M, Luttges J, Kloppel G, Jonat W, et al. Expression of the tumor suppressor gene maspin in human pancreatic cancers. Clin Cancer Res. 2001;7:812–7.PubMedGoogle Scholar
  48. 48.
    Maass N, Teffner M, Rosel F, Pawaresch R, Jonat W, Nagasaki K, et al. Decline in the expression of the serine proteinase inhibitor maspin is associated with tumour progression in ductal carcinomas of the breast. J Pathol. 2001;195:321–6.CrossRefPubMedGoogle Scholar
  49. 49.
    Xia W, Lau YK, Hu MC, Li L, Johnston DA, Sheng S, et al. High tumoral maspin expression is associated with improved survival of patients with oral squamous cell carcinoma. Oncogene. 2000;19:2398–403.CrossRefPubMedGoogle Scholar
  50. 50.
    Ferrucci PF, Rabascio C, Gigli F, Corsini C, Giordano G, Bertolini F, et al. A new comprehensive gene expression panel to study tumor micrometastasis in patients with high-risk breast cancer. Int J Oncol. 2007;30:955–62.PubMedGoogle Scholar
  51. 51.
    Joensuu KM, Leidenius MH, Andersson LC, Heikkila PS. High expression of maspin is associated with early tumor relapse in breast cancer. Hum Pathol. 2009;40:1143–51.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2010

Authors and Affiliations

  • Gayatri Sharma
    • 1
    • 4
  • Sameer Mirza
    • 1
  • Rajinder Parshad
    • 2
  • Anurag Srivastava
    • 2
  • Siddartha Datta Gupta
    • 3
  • Pranav Pandya
    • 4
  • Ranju Ralhan
    • 1
    • 5
    • 6
    • 7
    • 8
  1. 1.Department of BiochemistryAll India Institute of Medical SciencesAnsari NagarIndia
  2. 2.Department of SurgeryAll India Institute of Medical SciencesAnsari NagarIndia
  3. 3.Department of PathologyAll India Institute of Medical SciencesAnsari NagarIndia
  4. 4.Dev Sanskriti Vishwa VidhyalayaHardwarIndia
  5. 5.Department of Otolaryngology—Head and Neck SurgeryUniversity of TorontoTorontoCanada
  6. 6.Joseph and Mildred Sonshine Family Centre for Head and Neck DiseaseMount Sinai Hospital, Joseph and Wolf Lebovic Health ComplexTorontoCanada
  7. 7.Department of Otolaryngology—Head and Neck SurgeryMount Sinai Hospital, Joseph and Wolf Lebovic Health ComplexTorontoCanada
  8. 8.Department of Pathology and Laboratory MedicineMount Sinai Hospital, Joseph and Wolf Lebovic Health ComplexTorontoCanada

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