Skip to main content

Advertisement

SpringerLink
Log in

IGFBP7 reduces breast tumor growth by induction of senescence and apoptosis pathways

  • Preclinical study
  • Published:
Breast Cancer Research and Treatment Aims and scope Submit manuscript

Abstract

Insulin-like growth factor binding protein 7 (IGFBP7) has been shown to be a tumor suppressor in a variety of cancers. We previously have shown that IGFBP7 expression is inversely correlated with disease progression and poor outcome in breast cancer. Overexpression of IGFBP7 in MDA-MB-468, a triple-negative breast cancer (TNBC) cell line, resulted in inhibition of growth and migration. Xenografted tumors bearing ectopic IGFBP7 expression were significantly growth-impaired compared to IGFBP7-negative controls, which suggested that IGFBP7 treatment could inhibit breast cancer cell growth. To confirm this notion, 14 human patient primary breast tumors were analyzed by qRTPCR for IGFBP7 expression. The TNBC tumors expressed the lowest levels of IGFBP7 expression, which also correlated with higher tumorigenicity in mice. Furthermore, when breast cancer cell lines were treated with IGFBP7, only the TNBC cell lines were growth inhibited. Treatment of NOD/SCID mice harboring xenografts of TNBC cells with IGFBP7 systemically every 3–4 days inhibited tumorigenesis, with associated anti-angiogenic effects, together with increased apoptosis. Upon examining the mechanism of IGFBP7-mediated growth inhibition in TNBC cells, we found that cells not only were arrested in G1 phase of the cell cycle but also underwent senescence as a result of treatment with IGFBP7. Interestingly, IGFBP7 treatment was also associated with strong activation of the stress-associated p38 MAPK pathway, together with upregulation of p53 and the cyclin-dependent protein kinase (CDK) inhibitor, p21cip1. Prolonged treatment of cells with IGFBP7 resulted in increased cell death, marked by an increase in apoptotic cells and associated cleaved PARP. This is the first study showing that exogenous IGFBP7 inhibits TNBC cell growth both in vitro and in vivo. Taken together, these results suggest IGFBP7 treatment might have therapeutic potential for TNBC.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Wajapeyee N, Serra RW, Zhu X, Mahalingam M, Green MR (2008) Oncogenic BRAF induces senescence and apoptosis through pathways mediated by the secreted protein IGFBP7. Cell 132:363–374

    Article  PubMed  CAS  Google Scholar 

  2. Pollock PM, Harper UL, Hansen KS, Yudt LM, Stark M, Robbins CM, Moses TY, Hostetter G, Wagner U, Kakareka J, Salem G, Pohida T, Heenan P, Duray P, Kallioniemi O, Hayward NK, Trent JM, Meltzer PS (2003) High frequency of BRAF mutations in nevi. Nat Genet 33:19–20

    Article  PubMed  CAS  Google Scholar 

  3. Bauer J, Curtin JA, Pinkel D, Bastian BC (2007) Congenital melanocytic nevi frequently harbor NRAS mutations but no BRAF mutations. J Invest Dermatol 127:179–182

    Article  PubMed  CAS  Google Scholar 

  4. Papp T, Pemsel H, Zimmermann R, Bastrop R, Weiss DG, Schiffmann D (1999) Mutational analysis of the N-ras, p53, p16INK4a, CDK4, and MC1R genes in human congenital melanocytic naevi. J Med Genet 36:610–614

    PubMed  CAS  Google Scholar 

  5. Wajapeyee N, Kapoor V, Mahalingam M, Green MR (2009) Efficacy of IGFBP7 for treatment of metastatic melanoma and other cancers in mouse models and human cell lines. Mol Cancer Ther 8:3009–3014

    Article  PubMed  CAS  Google Scholar 

  6. Chen Y, Pacyna-Gengelbach M, Ye F, Knosel T, Lund P, Deutschmann N, Schluns K, Kotb WF, Sers C, Yasumoto H, Usui T, Petersen I (2007) Insulin-like growth factor binding protein-related protein 1 (IGFBP-rP1) has potential tumor-suppressive activity in human lung cancer. J Pathol 211:431–438

    Article  PubMed  CAS  Google Scholar 

  7. Ruan W, Xu E, Xu F, Ma Y, Deng H, Huang Q, Lv B, Hu H, Lin J, Cui J, Di M, Dong J, Lai M (2007) IGFBP7 plays a potential tumor suppressor role in colorectal carcinogenesis. Cancer Biol Ther 6:354–359

    Article  PubMed  CAS  Google Scholar 

  8. Sprenger CC, Damon SE, Hwa V, Rosenfeld RG, Plymate SR (1999) Insulin-like growth factor binding protein-related protein 1 (IGFBP-rP1) is a potential tumor suppressor protein for prostate cancer. Cancer Res 59:2370–2375

    PubMed  CAS  Google Scholar 

  9. Vizioli MG, Sensi M, Miranda C, Cleris L, Formelli F, Anania MC, Pierotti MA, Greco A (2010) IGFBP7: an oncosuppressor gene in thyroid carcinogenesis. Oncogene 29:3835–3844

    Article  PubMed  CAS  Google Scholar 

  10. Landberg G, Ostlund H, Nielsen NH, Roos G, Emdin S, Burger AM, Seth A (2001) Downregulation of the potential suppressor gene IGFBP-rP1 in human breast cancer is associated with inactivation of the retinoblastoma protein, cyclin E overexpression and increased proliferation in estrogen receptor negative tumors. Oncogene 20:3497–3505

    Article  PubMed  CAS  Google Scholar 

  11. Burger AM, Zhang X, Li H, Ostrowski JL, Beatty B, Venanzoni M, Papas T, Seth A (1998) Down-regulation of T1A12/mac25, a novel insulin-like growth factor binding protein related gene, is associated with disease progression in breast carcinomas. Oncogene 16:2459–2467

    Article  PubMed  CAS  Google Scholar 

  12. Burger AM, Zhang X, Seth A (1998) Detection of novel genes that are up-regulated (Di12) or down-regulated (T1A12) with disease progression in breast cancer. Eur J Cancer Prev 7(Suppl 1):S29–S35

    Article  PubMed  Google Scholar 

  13. Landberg G, Ostlund H, Nielsen NH, Roos G, Emdin S, Burger AM, Seth A (2001) Downregulation of the potential suppressor gene IGFBP-rP1 in human breast cancer is associated with inactivation of the retinoblastoma protein, cyclin E overexpression and increased proliferation in estrogen receptor negative tumors. Oncogene 20:3497–3505

    Article  PubMed  CAS  Google Scholar 

  14. Seth A, Kitching R, Landberg G, Xu J, Zubovits J, Burger AM (2003) Gene expression profiling of ductal carcinomas in situ and invasive breast tumors. Anticancer Res 23:2043–2051

    PubMed  CAS  Google Scholar 

  15. Burger A, Li H, Zhang XK, Pienkowska M, Venanzoni M, Vournakis J, Papas T, Seth A (1998) Breast cancer genome anatomy: correlation of morphological changes in breast carcinomas with expression of the novel gene product Di12. Oncogene 16:327–333

    Article  PubMed  CAS  Google Scholar 

  16. Burger AM, Zhang X, Li H, Ostrowski JL, Beatty B, Venanzoni M, Papas T, Seth A (1998) Down-regulation of T1A12/mac25, a novel insulin-like growth factor binding protein related gene, is associated with disease progression in breast carcinomas. Oncogene 16:2459–2467

    Article  PubMed  CAS  Google Scholar 

  17. Burger AM, Beatty B, Seth A (1999) Function and regulation of novel breast cancer associated genes. J Cancer Res Clin Oncol 125:S15–S16

    Google Scholar 

  18. Amemiya Y, Yang W, Benatar T, Nofech-Mozes S, Yee A, Kahn H, Holloway C, Seth A (2011) Insulin like growth factor binding protein-7 reduces growth of human breast cancer cells and xenografted tumors. Breast Cancer Res Treat 126:373–384

    Article  PubMed  CAS  Google Scholar 

  19. Dent R, Trudeau M, Pritchard KI, Hanna WM, Kahn HK, Sawka CA, Lickley LA, Rawlinson E, Sun P, Narod SA (2007) Triple-negative breast cancer: clinical features and patterns of recurrence. Clin Cancer Res 13:4429–4434

    Article  PubMed  Google Scholar 

  20. Kang Y, Siegel PM, Shu W, Drobnjak M, Kakonen SM, Cordon-Cardo C, Guise TA, Massague J (2003) A multigenic program mediating breast cancer metastasis to bone. Cancer Cell 3:537–549

    Article  PubMed  CAS  Google Scholar 

  21. Kwong J, Hong L, Liao R, Deng Q, Han J, Sun P (2009) p38alpha and p38gamma mediate oncogenic ras-induced senescence through differential mechanisms. J Biol Chem 284:11237–11246

    Article  PubMed  CAS  Google Scholar 

  22. Chen RY, Chen HX, Lin JX, She WB, Jiang P, Xu L, Tu YT (2010) In vivo transfection of pcDNA3.1-IGFBP7 inhibits melanoma growth in mice through apoptosis induction and VEGF downexpression. J Exp Clin Cancer Res 29:13

    Article  PubMed  Google Scholar 

  23. Inda AM, Andrini LB, Garcia MN, Garcia AL, Fernandez BA, Furnus CC, Galletti SM, Prat GD, Errecalde AL (2007) Evaluation of angiogenesis with the expression of VEGF and CD34 in human non-small cell lung cancer. J Exp Clin Cancer Res 26:375–378

    PubMed  CAS  Google Scholar 

  24. Sun T, Cao H, Xu L, Zhu B, Gu Q, Xu X (2011) Insulin-like growth factor binding protein-related protein 1 mediates VEGF-induced proliferation, migration and tube formation of retinal endothelial cells. Curr Eye Res 36:341–349

    Article  PubMed  Google Scholar 

  25. Tamura K, Hashimoto K, Suzuki K, Yoshie M, Kutsukake M, Sakurai T (2009) Insulin-like growth factor binding protein-7 (IGFBP7) blocks vascular endothelial cell growth factor (VEGF)-induced angiogenesis in human vascular endothelial cells. Eur J Pharmacol 610:61–67

    Article  PubMed  CAS  Google Scholar 

  26. Lin J, Lai M, Huang Q, Ruan W, Ma Y, Cui J (2008) Reactivation of IGFBP7 by DNA demethylation inhibits human colon cancer cell growth in vitro. Cancer Biol Ther 7:1896–1900

    Article  PubMed  CAS  Google Scholar 

  27. Hollestelle A, Elstrodt F, Nagel JH, Kallemeijn WW, Schutte M (2007) Phosphatidylinositol-3-OH kinase or RAS pathway mutations in human breast cancer cell lines. Mol Cancer Res 5:195–201

    Article  PubMed  CAS  Google Scholar 

  28. Bos JL (1989) Ras oncogenes in human cancer: a review. Cancer Res 49:4682–4689

    PubMed  CAS  Google Scholar 

  29. Hahn WC, Counter CM, Lundberg AS, Beijersbergen RL, Brooks MW, Weinberg RA (1999) Creation of human tumour cells with defined genetic elements. Nature 400:464–468

    Article  PubMed  CAS  Google Scholar 

  30. Rangarajan A, Hong SJ, Gifford A, Weinberg RA (2004) Species- and cell type-specific requirements for cellular transformation. Cancer Cell 6:171–183

    Article  PubMed  CAS  Google Scholar 

  31. Kamijo T, Zindy F, Roussel MF, Quelle DE, Downing JR, Ashmun RA, Grosveld G, Sherr CJ (1997) Tumor suppression at the mouse INK4a locus mediated by the alternative reading frame product p19ARF. Cell 91:649–659

    Article  PubMed  CAS  Google Scholar 

  32. Serrano M, Lin AW, McCurrach ME, Beach D, Lowe SW (1997) Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16INK4a. Cell 88:593–602

    Article  PubMed  CAS  Google Scholar 

  33. Serrano M, Blasco MA (2001) Putting the stress on senescence. Curr Opin Cell Biol 13:748–753

    Article  PubMed  CAS  Google Scholar 

  34. Campisi J (2005) Suppressing cancer: the importance of being senescent. Science 309:886–887

    Article  PubMed  CAS  Google Scholar 

  35. Bardeesy N, Sharpless NE (2006) RAS unplugged: negative feedback and oncogene-induced senescence. Cancer Cell 10:451–453

    Article  PubMed  CAS  Google Scholar 

  36. Lee AC, Fenster BE, Ito H, Takeda K, Bae NS, Hirai T, Yu ZX, Ferrans VJ, Howard BH, Finkel T (1999) Ras proteins induce senescence by altering the intracellular levels of reactive oxygen species. J Biol Chem 274:7936–7940

    Article  PubMed  CAS  Google Scholar 

  37. Hagen TM, Yowe DL, Bartholomew JC, Wehr CM, Do KL, Park JY, Ames BN (1997) Mitochondrial decay in hepatocytes from old rats: membrane potential declines, heterogeneity and oxidants increase. Proc Natl Acad Sci USA 94:3064–3069

    Article  PubMed  CAS  Google Scholar 

  38. Macip S, Igarashi M, Berggren P, Yu J, Lee SW, Aaronson SA (2003) Influence of induced reactive oxygen species in p53-mediated cell fate decisions. Mol Cell Biol 23:8576–8585

    Article  PubMed  CAS  Google Scholar 

  39. Polyak K, Xia Y, Zweier JL, Kinzler KW, Vogelstein B (1997) A model for p53-induced apoptosis. Nature 389:300–305

    Article  PubMed  CAS  Google Scholar 

  40. Macip S, Igarashi M, Fang L, Chen A, Pan ZQ, Lee SW, Aaronson SA (2002) Inhibition of p21-mediated ROS accumulation can rescue p21-induced senescence. EMBO J 21:2180–2188

    Article  PubMed  CAS  Google Scholar 

  41. Dolado I, Swat A, Ajenjo N, De VG, Cuadrado A, Nebreda AR (2007) p38alpha MAP kinase as a sensor of reactive oxygen species in tumorigenesis. Cancer Cell 11:191–205

    Article  PubMed  CAS  Google Scholar 

  42. Bulavin DV, Fornace AJ Jr (2004) p38 MAP kinase’s emerging role as a tumor suppressor. Adv Cancer Res 92:95–118

    Article  PubMed  CAS  Google Scholar 

  43. Bulavin DV, Saito S, Hollander MC, Sakaguchi K, Anderson CW, Appella E, Fornace AJ Jr (1999) Phosphorylation of human p53 by p38 kinase coordinates N-terminal phosphorylation and apoptosis in response to UV radiation. EMBO J 18:6845–6854

    Article  PubMed  CAS  Google Scholar 

  44. Huang C, Ma WY, Maxiner A, Sun Y, Dong Z (1999) p38 kinase mediates UV-induced phosphorylation of p53 protein at serine 389. J Biol Chem 274:12229–12235

    Article  PubMed  CAS  Google Scholar 

  45. Sun P, Yoshizuka N, New L, Moser BA, Li Y, Liao R, Xie C, Chen J, Deng Q, Yamout M, Dong MQ, Frangou CG, Yates JR III, Wright PE, Han J (2007) PRAK is essential for ras-induced senescence and tumor suppression. Cell 128:295–308

    Article  PubMed  CAS  Google Scholar 

  46. Sayed M, Kim SO, Salh BS, Issinger OG, Pelech SL (2000) Stress-induced activation of protein kinase CK2 by direct interaction with p38 mitogen-activated protein kinase. J Biol Chem 275:16569–16573

    Article  PubMed  CAS  Google Scholar 

  47. Shi Y, Gaestel M (2002) In the cellular garden of forking paths: how p38 MAPKs signal for downstream assistance. Biol Chem 383:1519–1536

    Article  PubMed  CAS  Google Scholar 

  48. Haq R, Brenton JD, Takahashi M, Finan D, Finkielsztein A, Damaraju S, Rottapel R, Zanke B (2002) Constitutive p38HOG mitogen-activated protein kinase activation induces permanent cell cycle arrest and senescence. Cancer Res 62:5076–5082

    PubMed  CAS  Google Scholar 

  49. Iwasa H, Han J, Ishikawa F (2003) Mitogen-activated protein kinase p38 defines the common senescence-signalling pathway. Genes Cells 8:131–144

    Article  PubMed  CAS  Google Scholar 

  50. Wang W, Chen JX, Liao R, Deng Q, Zhou JJ, Huang S, Sun P (2002) Sequential activation of the MEK-extracellular signal-regulated kinase and MKK3/6–p38 mitogen-activated protein kinase pathways mediates oncogenic ras-induced premature senescence. Mol Cell Biol 22:3389–3403

    Article  PubMed  Google Scholar 

  51. Chen Y, Pacyna-Gengelbach M, Ye F, Knosel T, Lund P, Deutschmann N, Schluns K, Kotb WF, Sers C, Yasumoto H, Usui T, Petersen I (2007) Insulin-like growth factor binding protein-related protein 1 (IGFBP-rP1) has potential tumour-suppressive activity in human lung cancer. J Pathol 211:431–438

    Article  PubMed  CAS  Google Scholar 

  52. Hwa V, Oh Y, Rosenfeld RG (1999) The insulin-like growth factor-binding protein (IGFBP) superfamily. Endocr Rev 20:761–787

    Article  PubMed  CAS  Google Scholar 

  53. Sprenger CC, Damon SE, Hwa V, Rosenfeld RG, Plymate SR (1999) Insulin-like growth factor binding protein-related protein 1 (IGFBP-rP1) is a potential tumor suppressor protein for prostate cancer. Cancer Res 59:2370–2375

    PubMed  CAS  Google Scholar 

  54. Sprenger CC, Vail ME, Evans K, Simurdak J, Plymate SR (2002) Over-expression of insulin-like growth factor binding protein-related protein-1(IGFBP-rP1/mac25) in the M12 prostate cancer cell line alters tumor growth by a delay in G1 and cyclin A associated apoptosis. Oncogene 21:140–147

    Article  PubMed  CAS  Google Scholar 

  55. Ingermann AR, Yang YF, Han J, Mikami A, Garza AE, Mohanraj L, Fan L, Idowu M, Ware JL, Kim HS, Lee DY, Oh Y (2010) Identification of a novel cell death receptor mediating IGFBP-3-induced anti-tumor effects in breast and prostate cancer. J Biol Chem 285:30233–30246

    Article  PubMed  CAS  Google Scholar 

  56. Lin J, Lai M, Huang Q, Ma Y, Cui J, Ruan W (2007) Methylation patterns of IGFBP7 in colon cancer cell lines are associated with levels of gene expression. J Pathol 212:83–90

    Article  PubMed  CAS  Google Scholar 

  57. Tomii K, Tsukuda K, Toyooka S, Dote H, Hanafusa T, Asano H, Naitou M, Doihara H, Kisimoto T, Katayama H, Pass HI, Date H, Shimizu N (2007) Aberrant promoter methylation of insulin-like growth factor binding protein-3 gene in human cancers. Int J Cancer 120:566–573

    Article  PubMed  CAS  Google Scholar 

  58. Ma Y, Lu B, Ruan W, Wang H, Lin J, Hu H, Deng H, Huang Q, Lai M (2008) Tumor suppressor gene insulin-like growth factor binding protein-related protein 1 (IGFBP-rP1) induces senescence-like growth arrest in colorectal cancer cells. Exp Mol Pathol 85:141–145

    Article  PubMed  CAS  Google Scholar 

  59. Swisshelm K, Ryan K, Tsuchiya K, Sager R (1995) Enhanced expression of an insulin growth factor-like binding protein (mac25) in senescent human mammary epithelial cells and induced expression with retinoic acid. Proc Natl Acad Sci USA 92:4472–4476

    Article  PubMed  CAS  Google Scholar 

  60. Goldstein S, Moerman EJ, Jones RA, Baxter RC (1991) Insulin-like growth factor binding protein 3 accumulates to high levels in culture medium of senescent and quiescent human fibroblasts. Proc Natl Acad Sci USA 88:9680–9684

    Article  PubMed  CAS  Google Scholar 

  61. Ahmed S, Yamamoto K, Sato Y, Ogawa T, Herrmann A, Higashi S, Miyazaki K (2003) Proteolytic processing of IGFBP-related protein-1 (TAF/angiomodulin/mac25) modulates its biological activity. Biochem Biophys Res Commun 310:612–618

    Article  PubMed  CAS  Google Scholar 

  62. Lalou C, Lassarre C, Binoux M (1996) A proteolytic fragment of insulin-like growth factor (IGF) binding protein-3 that fails to bind IGFs inhibits the mitogenic effects of IGF-I and insulin. Endocrinology 137:3206–3212

    Article  PubMed  CAS  Google Scholar 

  63. Wilson EM, Oh Y, Hwa V, Rosenfeld RG (2001) Interaction of IGF-binding protein-related protein 1 with a novel protein, neuroendocrine differentiation factor, results in neuroendocrine differentiation of prostate cancer cells. J Clin Endocrinol Metab 86:4504–4511

    Article  PubMed  CAS  Google Scholar 

  64. Schedlich LJ, Young TF, Firth SM, Baxter RC (1998) Insulin-like growth factor-binding protein (IGFBP)-3 and IGFBP-5 share a common nuclear transport pathway in T47D human breast carcinoma cells. J Biol Chem 273:18347–18352

    Article  PubMed  CAS  Google Scholar 

  65. Lee KW, Liu B, Ma L, Li H, Bang P, Koeffler HP, Cohen P (2004) Cellular internalization of insulin-like growth factor binding protein-3: distinct endocytic pathways facilitate re-uptake and nuclear localization. J Biol Chem 279:469–476

    Article  PubMed  CAS  Google Scholar 

  66. Santer FR, Bacher N, Moser B, Morandell D, Ressler S, Firth SM, Spoden GA, Sergi C, Baxter RC, Jansen-Durr P, Zwerschke W (2006) Nuclear insulin-like growth factor binding protein-3 induces apoptosis and is targeted to ubiquitin/proteasome-dependent proteolysis. Cancer Res 66:3024–3033

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Stephanie Bacopulos for editorial assistance. This work was supported by a CIHR grant, #MOP-97996 to Arun Seth.

Author information

Authors and Affiliations

  1. Division of Molecular and Cellular Biology, Sunnybrook Research Institute, 2075 Bayview Avenue, S-238, Toronto, ON, M4N 3M5, Canada

    Tania Benatar, Wenyi Yang, Yutaka Amemiya, Valentina Evdokimova & Arun Seth

  2. Department of Anatomic Pathology, Sunnybrook Health Science Center, Toronto, ON, Canada

    Harriette Kahn & Arun Seth

  3. Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON, Canada

    Arun Seth

  4. Genomics Core Facility, Sunnybrook Research Institute, Toronto, ON, Canada

    Yutaka Amemiya & Arun Seth

  5. Department of General Surgery, Odette Cancer Center, Sunnybrook Health Science Center, Toronto, ON, Canada

    Claire Holloway

  6. Institute of Protein Research, Pushchino, Moscow Region, 142290, Russia

    Valentina Evdokimova

Authors
  1. Tania Benatar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wenyi Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yutaka Amemiya
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Valentina Evdokimova
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Harriette Kahn
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Claire Holloway
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Arun Seth
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Arun Seth.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Benatar, T., Yang, W., Amemiya, Y. et al. IGFBP7 reduces breast tumor growth by induction of senescence and apoptosis pathways. Breast Cancer Res Treat 133, 563–573 (2012). https://doi.org/10.1007/s10549-011-1816-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10549-011-1816-4

Keywords

Search

Navigation