The broad-spectrum metalloproteinase inhibitor BB-94 inhibits growth, HER3 and Erk activation in fulvestrant-resistant breast cancer cell lines

Kirkegaard,Tove; Yde,Christina W.; Kveiborg,Marie; Lykkesfeldt,Anne E.

doi:10.3892/ijo.2014.2434

The broad-spectrum metalloproteinase inhibitor BB-94 inhibits growth, HER3 and Erk activation in fulvestrant-resistant breast cancer cell lines

Authors:
- Tove Kirkegaard
- Christina W. Yde
- Marie Kveiborg
- Anne E. Lykkesfeldt
View Affiliations

Affiliations: Breast Cancer Group, Cell Death and Metabolism, Danish Cancer Society Research Center, Copenhagen, Denmark, Department of Biomedical Sciences and Biotech Research and Innovation Centre (BRIC), University of Copenhagen, Copenhagen, Denmark
Published online on: May 9, 2014 https://doi.org/10.3892/ijo.2014.2434
Pages: 393-400

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Breast cancer cells can switch from estrogen receptor α (ER)- to human epidermal growth factor receptor (HER)-driven cell growth upon acquiring antiestrogen resistance. HER ligands are cleaved by metalloproteinases leading to release of active HER ligands, activation of HER receptors and consequently increased cell growth. In this study, we investigated the importance of HER receptors, in particular HER3, and HER ligand shedding for growth and signaling in human MCF-7 breast cancer cells and MCF-7-derived sublines resistant to the antiestrogen fulvestrant. The HER3/HER4 ligand heregulin 1β induced phosphorylation of HER3, Akt and Erk, and partly rescued fulvestrant-inhibited growth of MCF-7 cells. HER3 ligands were found to be produced and shed from the fulvestrant-resistant cells as conditioned medium from fulvestrant-resistant MCF-7 cells induced phosphorylation of HER3 and Akt in MCF-7 cells. This was prevented by treatment of resistant cells with the metalloproteinase inhibitor TAPI-2. Only the broad-spectrum metalloproteinase inhibitor BB-94, and not the more selective inhibitors GM6001 or TAPI-2, which inhibited shedding of the HER ligands produced by the fulvestrant-resistant cells, was able to inhibit growth and activation of HER3 and Erk in resistant cells. Compared to MCF-7, fulvestrant-resistant cells have increased HER3 phosphorylation, but knockdown of HER3 had no inhibitory effect on resistant cell growth. The EGFR inhibitor gefitinib exhibited only a minor growth inhibition, whereas the pan-HER inhibitor CI-1033 exerted growth arrest. Thus, neither HER3 nor EGFR alone are the main driver of fulvestrant-resistant cell growth and treatment should target both receptors. Ligand shedding is not a treatment target, as receptor activation occurred, independent of release of ligands. Only the broad-spectrum metalloproteinase inhibitor BB-94 could abrogate HER3 and Erk activation in the resistant cells, which stresses the complexity of the resistance mechanisms and the requirement of targeting signaling from HER receptors by multiple strategies.

View Figures

View References

1.	Valachis A, Mauri D, Polyzos NP, Mavroudis D, Georgoulias V and Casazza G: Fulvestrant in the treatment of advanced breast cancer: a systematic review and meta-analysis of randomized controlled trials. Crit Rev Oncol Hematol. 73:220–227. 2010. View Article : Google Scholar : PubMed/NCBI
2.	Frogne T, Benjaminsen RV, Sonne-Hansen K, et al: Activation of ErbB3, EGFR and Erk is essential for growth of human breast cancer cell lines with acquired resistance to fulvestrant. Breast Cancer Res Treat. 114:263–275. 2009. View Article : Google Scholar : PubMed/NCBI
3.	McClelland RA, Barrow D, Madden TA, et al: Enhanced epidermal growth factor receptor signaling in MCF7 breast cancer cells after long-term culture in the presence of the pure antiestrogen ICI 182,780 (Faslodex). Endocrinology. 142:2776–2788. 2001.PubMed/NCBI
4.	Sommer A, Hoffmann J, Lichtner RB, Schneider MR and Parczyk K: Studies on the development of resistance to the pure antiestrogen Faslodex in three human breast cancer cell lines. J Steroid Biochem Mol Biol. 85:33–47. 2003. View Article : Google Scholar : PubMed/NCBI
5.	Massarweh S, Osborne CK, Creighton CJ, et al: Tamoxifen resistance in breast tumors is driven by growth factor receptor signaling with repression of classic estrogen receptor genomic function. Cancer Res. 68:826–833. 2008. View Article : Google Scholar
6.	Sonne-Hansen K, Norrie IC, Emdal KB, et al: Breast cancer cells can switch between estrogen receptor alpha and ErbB signaling and combined treatment against both signaling pathways postpones development of resistance. Breast Cancer Res Treat. 121:601–613. 2010. View Article : Google Scholar
7.	Zahnow CA: ErbB receptors and their ligands in the breast. Expert Rev Mol Med. 8:1–21. 2006. View Article : Google Scholar : PubMed/NCBI
8.	Gullick WJ: c-erbB-4/HER4: friend or foe? J Pathol. 200:279–281. 2003. View Article : Google Scholar : PubMed/NCBI
9.	Yarden Y and Sliwkowski MX: Untangling the ErbB signalling network. Nat Rev Mol Cell Biol. 2:127–137. 2001. View Article : Google Scholar : PubMed/NCBI
10.	Witton CJ, Reeves JR, Going JJ, Cooke TG and Bartlett JM: Expression of the HER1-4 family of receptor tyrosine kinases in breast cancer. J Pathol. 200:290–297. 2003. View Article : Google Scholar : PubMed/NCBI
11.	Tovey S, Dunne B, Witton CJ, Forsyth A, Cooke TG and Bartlett JM: Can molecular markers predict when to implement treatment with aromatase inhibitors in invasive breast cancer? Clin Cancer Res. 11:4835–4842. 2005. View Article : Google Scholar : PubMed/NCBI
12.	Frogne T, Laenkholm AV, Lyng MB, Henriksen KL and Lykkesfeldt AE: Determination of HER2 phosphorylation at tyrosine 1221/1222 improves prediction of poor survival for breast cancer patients with hormone receptor-positive tumors. Breast Cancer Res. 11:R112009. View Article : Google Scholar : PubMed/NCBI
13.	Blobel CP: ADAMs: key components in EGFR signalling and development. Nat Rev Mol Cell Biol. 6:32–43. 2005. View Article : Google Scholar : PubMed/NCBI
14.	Piccart-Gebhart MJ, Procter M, Leyland-Jones B, et al: Trastuzumab after adjuvant chemotherapy in HER2-positive breast cancer. N Engl J Med. 353:1659–1672. 2005. View Article : Google Scholar : PubMed/NCBI
15.	Slamon DJ, Leyland-Jones B, Shak S, et al: Use of chemotherapy plus a monoclonal antibody against HER2 for metastatic breast cancer that overexpresses HER2. N Engl J Med. 344:783–792. 2001. View Article : Google Scholar : PubMed/NCBI
16.	Valabrega G, Montemurro F, Sarotto I, et al: TGFalpha expression impairs Trastuzumab-induced HER2 downregulation. Oncogene. 24:3002–3010. 2005. View Article : Google Scholar : PubMed/NCBI
17.	Motoyama AB, Hynes NE and Lane HA: The efficacy of ErbB receptor-targeted anticancer therapeutics is influenced by the availability of epidermal growth factor-related peptides. Cancer Res. 62:3151–8315. 2002.PubMed/NCBI
18.	Lykkesfeldt AE, Larsen SS and Briand P: Human breast cancer cell lines resistant to pure anti-estrogens are sensitive to tamoxifen treatment. Int J Cancer. 61:529–354. 1995. View Article : Google Scholar : PubMed/NCBI
19.	Frogne T, Jepsen JS, Larsen SS, Fog CK, Brockdorff BL and Lykkesfeldt AE: Antiestrogen-resistant human breast cancer cells require activated protein kinase B/Akt for growth. Endocr Relat Cancer. 12:599–614. 2005. View Article : Google Scholar : PubMed/NCBI
20.	Lundholt BK, Briand P and Lykkesfeldt AE: Growth inhibition and growth stimulation by estradiol of estrogen receptor transfected human breast epithelial cell lines involve different pathways. Breast Cancer Res Treat. 67:199–214. 2001. View Article : Google Scholar
21.	Pancholi S, Lykkesfeldt A, Hilmi C, et al: ERBB2 influences the subcellular localization of the estrogen receptor in tamoxifen-resistant MCF-7 cells leading to the activation of AKT and p90RSK. Endocr Relat Cancer. 15:985–1002. 2008. View Article : Google Scholar : PubMed/NCBI
22.	Nicholson RI, Hutcheson IR, Harper ME, et al: Modulation of epidermal growth factor receptor in endocrine-resistant, oestrogen receptor-positive breast cancer. Endocr Relat Cancer. 8:175–182. 2001. View Article : Google Scholar : PubMed/NCBI
23.	Rasmussen BB, Regan MM, Lykkesfeldt AE, et al: Adjuvant letrozole versus tamoxifen according to centrally-assessed ERBB2 status for postmenopausal women with endocrine-responsive early breast cancer: supplementary results from the BIG 1–98 randomised trial. Lancet Oncol. 9:23–28. 2008.
24.	Schwartzberg LS, Franco SX, Florance A, O'Rourke L, Maltzman J and Johnston S: Lapatinib plus letrozole as first-line therapy for HER-2⁺ hormone receptor-positive metastatic breast cancer. Oncologist. 15:122–129. 2010. View Article : Google Scholar : PubMed/NCBI
25.	Larsen SS, Egeblad M, Jaattela M and Lykkesfeldt AE: Acquired antiestrogen resistance in MCF-7 human breast cancer sublines is not accomplished by altered expression of receptors in the ErbB-family. Breast Cancer Res Treat. 58:41–56. 1999. View Article : Google Scholar : PubMed/NCBI
26.	Sahin U, Weskamp G, Kelly K, et al: Distinct roles for ADAM10 and ADAM17 in ectodomain shedding of six EGFR ligands. J Cell Biol. 164:769–779. 2004. View Article : Google Scholar : PubMed/NCBI