Skip to main content

Advertisement

SpringerLink
Log in

Germ line knockout of IGFBP-3 reveals influences of the gene on mammary gland neoplasia

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

Abstract

Insulin-like growth factor binding protein-3 (IGFBP-3) is an important carrier protein for insulin-like growth factors (IGFs) in the circulation. IGFBP-3 antagonizes the growth-promoting and anti-apoptotic activities of IGFs in experimental systems, but in certain contexts can increase IGF bioactivity, probably by increasing its half-life. The goal of this study was to investigate the role of IGFBP-3 in breast carcinogenesis and breast cancer metastasis. In the first part of the study, we exposed IGFBP-3 knockout and wild-type female mice to dimethylbenz[a]anthracene (DMBA) and followed them for appearance of primary tumors for up to 13 months. In the second part, mice of each genotype received an IV injection of 4T1 mammary carcinoma cells and then lung nodules were counted. Our results show that IGFBP-3 knockout mice developed breast tumors significantly earlier than the wild-type (13.9 ± 1.1 versus 22.5 ± 3.3 weeks, respectively, P = 0.0144), suggesting tumor suppression activity of IGFBP-3. In tumors of IGFBP-3 knockout mice, levels of phospho-AKTSer473 were increased compared to wild-type mice. The lung metastasis assay showed significantly more and larger lung nodules in IGFBP-3 knockout mice than in wild-type mice. While we observed increased levels of IGFBP-5 protein in the IGFBP-3 knockout mice, our findings suggest that this was not sufficient to completely compensate for the absence of IGFBP-3. Even though knockout of IGFBP-3 is associated with only a subtle phenotype under control conditions, our results reveal that loss of this gene has measurable effects on breast carcinogenesis and breast cancer metastasis.

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

Similar content being viewed by others

References

  1. Firth SM, Baxter RC (2002) Cellular actions of the insulin-like growth factor binding proteins. Endocr Rev 23:824–854

    Article  CAS  PubMed  Google Scholar 

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

    CAS  PubMed  Google Scholar 

  3. Jogie-Brahim S, Feldman D, Oh Y (2009) Unraveling insulin-like growth factor binding protein-3 actions in human disease. Endocr Rev 30(5):417–437

    Article  CAS  Google Scholar 

  4. Pollak MN, Schernhammer ES, Hankinson SE (2004) Insulin-like growth factors and neoplasia. Nat Rev Cancer 4(7):505–518

    Article  CAS  PubMed  Google Scholar 

  5. Lu Y, Zi X, Zhao Y, Mascarenhas D, Pollak M (2001) Insulin-like growth factor-I receptor signaling and resistance to trastuzumab (Herceptin). J Natl Cancer Inst 93:1852–1857

    Article  CAS  PubMed  Google Scholar 

  6. Nickerson T, Huynh H, Pollak M (1997) Insulin-like growth factor binding protein-3 induces apoptosis in MCF7 breast cancer cells. Biochem Biophys Res Commun 237:690–693

    Article  CAS  PubMed  Google Scholar 

  7. Pratt S, Pollak M (1994) Insulin-like growth factor binding protein 3 inhibits estrogen-stimulated breast cancer cell proliferation. Biochem Biophys Res Commun 198:292–297

    Article  CAS  PubMed  Google Scholar 

  8. Shen L, Dean NM, Glazer RI (1999) Induction of p53-dependent, insulin-like growth factor-binding protein-3-mediated apoptosis in glioblastoma multiforme cells by a protein kinase Calpha antisense oligonucleotide. Mol Pharmacol 55:396–402

    CAS  PubMed  Google Scholar 

  9. Butt AJ, Williams DC (2001) IGFBP-3 and apoptosis—a licence to kill? Apoptosis 6:199–205

    Article  CAS  PubMed  Google Scholar 

  10. Gill ZP, Perks CM, Newcomb PV, Holly JM (1997) Insulin-like growth factor-binding protein (IGFBP-3) predisposes breast cancer cells to programmed cell death in a non-IGF-dependent manner. J Biol Chem 272:25602–25607

    Article  CAS  PubMed  Google Scholar 

  11. Hong J, Zhang G, Dong F, Rechler MM (2002) Insulin-like growth factor (IGF)-binding protein-3 mutants that do not bind IGF-I or IGF-II stimulate apoptosis in human prostate cancer cells. J Biol Chem 277:10489–10497

    Article  CAS  PubMed  Google Scholar 

  12. McCaig C, Fowler CA, Laurence NJ, Lai T, Savage PB, Holly JMP, Perks CM (2002) Differential interactions between IGFBP-3 and transforming growth factor-beta (TGF-beta) in normal vs cancerous breast epithelial cells. Br J Cancer 86:1963–1969

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  13. Williams AC, Collard TJ, Perks CM, Newcomb P, Moorghen M, Holly JMP, Paraskeva C (2000) Increased p53-dependent apoptosis by the insulin-like growth factor binding protein IGFBP-3 in human colonic adenoma-derived cells. Cancer Res 60:22–27

    CAS  PubMed  Google Scholar 

  14. Liu B, Lee KW, Anzo M, Zhang B, Zi X, Tao Y, Shiry L, Pollak M, Lin S, Cohen P (2007) Insulin-like growth factor-binding protein-3 inhibition of prostate cancer growth involves suppression of angiogenesis. Oncogene 26:1811–1819

    Article  CAS  PubMed  Google Scholar 

  15. Gucev ZS, Oh Y, Kelley KM, Rosenfeld RG (1996) Insulin-like growth factor binding protein 3 mediates retinoic acid and transforming growth factor beta 2-induced growth inhibition in human breast cancer cells. Cancer Res 56:1545–1550

    CAS  PubMed  Google Scholar 

  16. Huynh HT, Robitaille G, Turner JD (1991) Establishment of bovine mammary epithelial cells (MAC-T): an in vitro model for bovine lactation. Exp Cell Res 197:191–199

    Article  CAS  PubMed  Google Scholar 

  17. Huynh HT, Yang XF, Pollak M (1996) Estradiol and antiestrogens regulate a growth inhibitory insulin-like growth factor binding protein 3 autocrine loop in human breast cancer cells. J Biol Chem 271:1016–1021

    Article  CAS  PubMed  Google Scholar 

  18. Khandwala HM, McCutcheon IE, Flyvbjerg A, Friend KE (2000) The effects of insulin-like growth factors on tumorigenesis and neoplastic growth. Endocr Rev 21:215–244

    Article  CAS  Google Scholar 

  19. Martin JL, Coverley JA, Pattison ST, Baxter RC (1995) Insulin-like growth factor-binding protein-3 production by MCF-7 breast cancer cells: stimulation by retinoic acid and cyclic adenosine monophosphate and differential effects of estradiol. Endocrinology 136:1219–1226

    CAS  PubMed  Google Scholar 

  20. Torrisi R, Parodi S, Fontana V, Pensa F, Casella C, Barreca A, De P, Costa A, Decensi A (1998) Effect of fenretinide on plasma IGF-I and IGFBP-3 in early breast cancer patients. Int J Cancer 76:787–790

    Article  CAS  PubMed  Google Scholar 

  21. Burrows C, Shiry LJ, Holly JMP, Perks CM (2003) Differential effects of IGFBP-3 on apoptosis of breast epithelial cells according to apoptotic trigger. Growth Hormone and IGF Research 13 (Abstract P28):203–222

  22. Fowler CA, Perks CM, Newcomb PV, Savage PB, Farndon JR, Holly JM (2000) Insulin-like growth factor binding protein-3 (IGFBP-3) potentiates paclitaxel-induced apoptosis in human breast cancer cells. Int J Cancer 88:448–453

    Article  CAS  PubMed  Google Scholar 

  23. Neuenschwander S, Schwartz A, Wood TL, Roberts CT Jr, Hennighausen L, LeRoith D (1996) Involution of the lactating mammary gland is inhibited by the IGF system in a transgenic mouse model. J Clin Investig 97(10):2225–2232

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  24. Schneider MR, Lahm H, Wu M, Hoeflich A, Wolf E (2000) Transgenic mouse models for studying the functions of insulin-like growth factor-binding proteins. FASEB J 14(5):629–640

    CAS  PubMed  Google Scholar 

  25. Silha JV, Murphy LJ (2002) Insights from insulin-like growth factor binding protein transgenic mice. Endocrinology 143(10):3711–3714

    Article  CAS  PubMed  Google Scholar 

  26. Silha JV, Sheppard PC, Mishra S, Gui Y, Schwartz J, Dodd JG, Murphy LJ (2006) Insulin-like growth factor (IGF) binding protein-3 attenuates prostate tumor growth by IGF-dependent and IGF-independent mechanisms. Endocrinology 147(5):2112–2121

    Article  CAS  PubMed  Google Scholar 

  27. Ning Y, Schuller AG, Bradshaw S, Rotwein P, Ludwig T, Frystyk J, Pintar JE (2006) Diminished growth and enhanced glucose metabolism in triple knockout mice containing mutations of insulin-like growth factor binding protein-3, -4, and -5. Mol Endocrinol 20(9):2173–2186

    Article  CAS  PubMed  Google Scholar 

  28. Mehta HH, Gao Q, Galet C, Paharkova V, Wan J, Said J, Sohn JJ, Lawson G, Cohen P, Cobb LJ, Lee KW (2011) IGFBP-3 is a metastasis suppression gene in prostate cancer. Cancer Res 71(15):5154–5163

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  29. Lue Y, Swerdloff R, Liu Q, Mehta H, Hikim AS, Lee KW, Jia Y, Hwang D, Cobb LJ, Cohen P, Wang C (2010) Opposing roles of insulin-like growth factor binding protein 3 and humanin in the regulation of testicular germ cell apoptosis. Endocrinology 151(1):350–357

    Article  CAS  PubMed  Google Scholar 

  30. Aldaz CM, Liao QY, LaBate M, Johnston DA (1996) Medroxyprogesterone acetate accelerates the development and increases the incidence of mouse mammary tumors induced by dimethylbenzanthracene. Carcinogenesis 17(9):2069–2072

    Article  CAS  PubMed  Google Scholar 

  31. Siddiqui RA, Harvey KA, Walker C, Altenburg J, Xu Z, Terry C, Camarillo I, Jones-Hall Y, Mariash C (2013) Characterization of synergistic anti-cancer effects of docosahexaenoic acid and curcumin on DMBA-induced mammary tumorigenesis in mice. BMC Cancer 13:418. doi:10.1186/1471-2407-13-418

    Article  PubMed Central  PubMed  Google Scholar 

  32. Gribben L, Baxter RC, Marsh DJ (2012) Insulin-like growth factor binding protein-3 inhibits migration of endometrial cancer cells. Cancer Lett 317(1):41–48. doi:10.1016/j.canlet.2011.11.011

    Article  CAS  PubMed  Google Scholar 

  33. Chun SY, Chen F, Washburn JG, MacDonald JW, Innes KL, Zhao R, Cruz-Correa MR, Dang LH, Dang DT (2007) CDX2 promotes anchorage-independent growth by transcriptional repression of IGFBP-3. Oncogene 26(32):4725–4729. doi:10.1038/sj.onc.1210258

    Article  CAS  PubMed  Google Scholar 

  34. Saldana SM, Lee HH, Lowery FJ, Khotskaya YB, Xia W, Zhang C, Chang SS, Chou CK, Steeg PS, Yu D, Hung MC (2013) Inhibition of type I insulin-like growth factor receptor signaling attenuates the development of breast cancer brain metastasis. PLoS One 8(9):e73406. doi:10.1371/journal.pone.0073406

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  35. Gahete MD, Cordoba-Chacon J, Lantvit DD, Ortega-Salas R, Sanchez-Sanchez R, Perez-Jimenez F, Lopez-Miranda J, Swanson SM, Castano JP, Luque RM, Kineman RD (2014) Elevated GH/IGF-I promotes mammary tumors in high-fat, but not low-fat, fed mice. Carcinogenesis. doi:10.1093/carcin/bgu161

    PubMed  Google Scholar 

  36. Pollak M, Blouin MJ, Zhang JC, Kopchick JJ (2001) Reduced mammary gland carcinogenesis in transgenic mice expressing a growth hormone antagonist. Br J Cancer 85(3):428–430. doi:10.1054/bjoc.2001.1895

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  37. Ramsey MM, Ingram RL, Cashion AB, Ng AH, Cline JM, Parlow AF, Sonntag WE (2002) Growth hormone-deficient dwarf animals are resistant to dimethylbenzanthracine (DMBA)-induced mammary carcinogenesis. Endocrinology 143(10):4139–4142. doi:10.1210/en.2002-220717

    Article  CAS  PubMed  Google Scholar 

  38. Deal C, Ma J, Wilkin F, Paquette J, Rozen F, Ge B, Hudson T, Stampfer M, Pollak M (2001) Novel promoter polymorphism in insulin-like growth factor-binding protein-3: correlation with serum levels and interaction with known regulators. J Clin Endocrinol Metab 86(3):1274–1280

    CAS  PubMed  Google Scholar 

  39. Jernstrom H, Wilkin F, Deal C, Chu W, Tao Y, Majeed N, Narod S, Hudson T, Pollak M (2001) Genetic and non-genetic factors associated with variation of plasma levels of insulin-like growth factor-I and insulin-like growth factor binding protein-3 in healthy premenopausal women. Cancer Epidemiol Biomark Prev 10:377–384

    CAS  Google Scholar 

Download references

Acknowledgments

We would like to thank Ms. Véronique Michaud and Mr. Yvhans Chery for their technical assistance with the animal care, Dr. Naciba Benlimame and Ms. Lilian Canetti for their collaboration for the immunohistochemistry procedures and Ms Jasmin Sander for the scanning of the slides. We are also grateful to Ms. Rhoda Lim for her help in the submission of this manuscript.

Conflict of interest

Dr. David R. Powell is the Vice-President of Metabolism Research at Lexicon Pharmaceuticals, Inc., of which the knockout mice were generated. The other authors have no conflict of interest to disclose.

Funding

Miguel Bazile was supported through the McGill Integrated Cancer Research Training Program Studentship.

Author information

Authors and Affiliations

  1. Lady Davis Institute for Medical Research of the Jewish General Hospital, Room E-423, 3755 Cote-Ste-Catherine Road, Montreal, QC, H3T 1E2, Canada

    Marie-José Blouin, Miguel Bazile, Elena Birman, Mahvash Zakikhani & Michael Pollak

  2. Department of Pathology, McGill University Health Center, McGill University, Montreal, QC, Canada

    Livia Florianova

  3. Department of Pathology, Jewish General Hospital, Montreal, QC, Canada

    Olga Aleynikova

  4. Lexicon Pharmaceuticals Inc., The Woodlands, TX, USA

    David R. Powell

  5. Department of Oncology, McGill University, Montreal, QC, Canada

    Michael Pollak

Authors
  1. Marie-José Blouin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Miguel Bazile
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Elena Birman
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mahvash Zakikhani
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Livia Florianova
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Olga Aleynikova
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. David R. Powell
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Michael Pollak
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Marie-José Blouin.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Blouin, MJ., Bazile, M., Birman, E. et al. Germ line knockout of IGFBP-3 reveals influences of the gene on mammary gland neoplasia. Breast Cancer Res Treat 149, 577–585 (2015). https://doi.org/10.1007/s10549-015-3268-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10549-015-3268-8

Keywords

Search

Navigation