Top

Published in:

01-06-2009 | Preclinical Study

Cyclopamine inhibition of human breast cancer cell growth independent of Smoothened (Smo)

Authors: Xiaomei Zhang, Nikesha Harrington, Ricardo C. Moraes, Meng-Fen Wu, Susan G. Hilsenbeck, Michael T. Lewis

Published in: Breast Cancer Research and Treatment | Issue 3/2009

Abstract

Altered hedgehog signaling is implicated in the development of approximately 20–25% of all cancers, especially those of soft tissues. Genetic evidence in mice as well as immunolocalization studies in human breast cancer specimens suggest that deregulated hedgehog signaling may contribute to breast cancer development. Indeed, two recent studies demonstrated that anchorage-dependent growth of some human breast cancer cell lines is impaired by cyclopamine, a potent hedgehog signaling antagonist targeting the Smoothened (SMO) protein. However, specificity of cyclopamine at the dosage required for growth inhibition (≥10 μM) remained an open question. In this paper we demonstrate that hedgehog signaling antagonists, including cyclopamine, and a second compound, CUR0199691, can inhibit growth of estrogen receptor (ER)-positive and ER-negative tumorigenic breast cancer cells at elevated doses. However, our results indicate that, for most breast cancer cell lines, growth inhibition by these compounds can be independent of detectable Smo gene expression. Rather, our results suggest that cyclopamine and CUR0199691 have unique secondary molecular targets at the dosages required for growth inhibition that are unrelated to hedgehog signaling.

Available only for authorised users

Lewis MT, Veltmaat JM (2004) Next stop, the twilight zone: hedgehog network regulation of mammary gland development. J Mammary Gland Biol Neoplasia 9:165–181. doi:10.1023/B:JOMG.0000037160.24731.35 PubMedCrossRef

Hooper JE, Scott MP (2005) Communicating with hedgehogs. Nat Rev Mol Cell Biol 6:306–317. doi:10.1038/nrm1622 PubMedCrossRef

Nusse R (2003) Wnts and hedgehogs: lipid-modified proteins and similarities in signaling mechanisms at the cell surface. Development 130:5297–5305. doi:10.1242/dev.00821 PubMedCrossRef

Cohen MM Jr (2003) The hedgehog signaling network. Am J Med Genet 123A:5–28. doi:10.1002/ajmg.a.20495 CrossRefPubMed

Liu S, Dontu G, Mantle ID et al (2006) Hedgehog signaling and Bmi-1 regulate self-renewal of normal and malignant human mammary stem cells. Cancer Res 66:6063–6071. doi:10.1158/0008-5472.CAN-06-0054 PubMedCrossRef

Briscoe J, Therond P (2005) Hedgehog signaling: from the Drosophila cuticle to anti-cancer drugs. Dev Cell 8:143–151. doi:10.1016/j.devcel.2005.01.008 PubMedCrossRef

Chang-Claude J, Dunning A, Schnitzbauer U et al (2003) The patched polymorphism Pro1315Leu (C3944T) may modulate the association between use of oral contraceptives and breast cancer risk. Int J Cancer 103:779–783. doi:10.1002/ijc.10889 PubMedCrossRef

Naylor TL, Greshock J, Wang Y et al (2005) High resolution genomic analysis of sporadic breast cancer using array-based comparative genomic hybridization. Breast Cancer Res 7:R1186–R1198. doi:10.1186/bcr1356 PubMedCrossRef

Kubo M, Nakamura M, Tasaki A et al (2004) Hedgehog signaling pathway is a new therapeutic target for patients with breast cancer. Cancer Res 64:6071–6074. doi:10.1158/0008-5472.CAN-04-0416 PubMedCrossRef

10.

Mukherjee S, Frolova N, Sadlonova A et al (2006) Hedgehog signaling and response to cyclopamine differ in epithelial and stromal cells in benign breast and breast cancer. Cancer Biol Ther 5:674–683PubMedCrossRef

11.

Wolf I, Bose S, Desmond JC et al (2007) Unmasking of epigenetically silenced genes reveals DNA promoter methylation and reduced expression of PTCH in breast cancer. Breast Cancer Res Treat 105:139–155PubMedCrossRef

12.

Moraes RC, Zhang X, Harrington N et al (2007) Constitutive activation of smoothened (Smo) in mammary glands of transgenic mice leads to increased proliferation, altered differentiation and ductal dysplasia. Development 134:1231–1242 PubMedCrossRef

13.

Xie J, Johnson RL, Zhang X et al (1997) Mutations of the PATCHED gene in several types of sporadic extracutaneous tumors. Cancer Res 57:2369–2372PubMed

14.

Evangelista M, Tian H, de Sauvage FJ (2006) The hedgehog signaling pathway in cancer. Clin Cancer Res 12:5924–5928. doi:10.1158/1078-0432.CCR-06-1736 PubMedCrossRef

15.

Riobo NA, Saucy B, Dilizio C, Manning DR (2006) Activation of heterotrimeric G proteins by Smoothened. Proc Natl Acad Sci USA 103:12607–12612. doi:10.1073/pnas.0600880103 PubMedCrossRef

16.

Barnes EA, Kong M, Ollendorff V, Donoghue DJ (2001) Patched1 interacts with cyclin B1 to regulate cell cycle progression. EMBO J 20:2214–2223. doi:10.1093/emboj/20.9.2214 PubMedCrossRef

17.

Meloni AR, Fralish GB, Kelly P et al (2006) Smoothened signal transduction is promoted by g protein-coupled receptor kinase 2. Mol Cell Biol 26:7550–7560. doi:10.1128/MCB.00546-06 PubMedCrossRef

18.

Chen W, Ren XR, Nelson CD et al (2004) Activity-dependent internalization of smoothened mediated by beta-arrestin 2 and GRK2. Science 306:2257–2260. doi:10.1126/science.1104135 PubMedCrossRef

19.

Frank-Kamenetsky M, Zhang XM, Bottega S et al (2002) Small-molecule modulators of hedgehog signaling: identification and characterization of Smoothened agonists and antagonists. J Biol 1:10. doi:10.1186/1475-4924-1-10 PubMedCrossRef

20.

Williams JA, Guicherit OM, Zaharian BI et al (2003) Identification of a small molecule inhibitor of the hedgehog signaling pathway: effects on basal cell carcinoma-like lesions. Proc Natl Acad Sci USA 100:4616–4621. doi:10.1073/pnas.0732813100 PubMedCrossRef

21.

Chen JK, Taipale J, Young KE, Maiti T, Beachy PA (2002) Small molecule modulation of Smoothened activity. Proc Natl Acad Sci USA 99:14071–14076. doi:10.1073/pnas.182542899 PubMedCrossRef

22.

Incardona JP, Gaffield W, Kapur RP, Roelink H (1998) The teratogenic Veratrum alkaloid cyclopamine inhibits sonic hedgehog signal transduction. Development 125:3553–3562PubMed

23.

Gabay L, Lowell S, Rubin LL, Anderson DJ (2003) Deregulation of dorsoventral patterning by FGF confers trilineage differentiation capacity on CNS stem cells in vitro. Neuron 40:485–499. doi:10.1016/S0896-6273(03)00637-8 PubMedCrossRef

24.

Levitt RJ, Zhao Y, Blouin MJ, Pollak M (2007) The hedgehog pathway inhibitor cyclopamine increases levels of p27, and decreases both expression of IGF-II and activation of Akt in PC-3 prostate cancer cells. Cancer Lett 255:300–306PubMedCrossRef

25.

Masdeu C, Faure H, Coulombe J et al (2006) Identification and characterization of hedgehog modulator properties after functional coupling of Smoothened to G15. Biochem Biophys Res Commun 349:471–479. doi:10.1016/j.bbrc.2006.07.216 PubMedCrossRef

26.

Mimeault M, Moore E, Moniaux N et al (2005) Cytotoxic effects induced by a combination of cyclopamine and gefitinib, the selective hedgehog and epidermal growth factor receptor signaling inhibitors, in prostate cancer cells. Int J Cancer 118:1022–1031CrossRef

27.

Chen JK, Taipale J, Cooper MK, Beachy PA (2002) Inhibition of hedgehog signaling by direct binding of cyclopamine to Smoothened. Genes Dev 16:2743–2748. doi:10.1101/gad.1025302 PubMedCrossRef

28.

Berman DM, Karhadkar SS, Hallahan AR et al (2002) Medulloblastoma growth inhibition by hedgehog pathway blockade. Science 297:1559–1561. doi:10.1126/science.1073733 PubMedCrossRef

29.

Tabs S, Avci O (2004) Induction of the differentiation and apoptosis of tumor cells in vivo with efficiency and selectivity. Eur J Dermatol 14:96–102PubMed

30.

Sanchez P, Ruiz i Altaba A (2005) In vivo inhibition of endogenous brain tumors through systemic interference of Hedgehog signaling in mice. Mech Dev 122:223–230. doi:10.1016/j.mod.2004.10.002 PubMedCrossRef

31.

Athar M, Li C, Tang X et al (2004) Inhibition of smoothened signaling prevents ultraviolet B-induced basal cell carcinomas through regulation of Fas expression and apoptosis. Cancer Res 64:7545–7552. doi:10.1158/0008-5472.CAN-04-1393 PubMedCrossRef

32.

Romer JT, Kimura H, Magdaleno S et al (2004) Suppression of the Shh pathway using a small molecule inhibitor eliminates medulloblastoma in Ptc1(+/−)p53(−/−) mice. Cancer Cell 6:229–240. doi:10.1016/j.ccr.2004.08.019 PubMedCrossRef

33.

Sasai K, Romer JT, Lee Y et al (2006) Shh pathway activity is down-regulated in cultured medulloblastoma cells: implications for preclinical studies. Cancer Res 66:4215–4222. doi:10.1158/0008-5472.CAN-05-4505 PubMedCrossRef

34.

Lewis DE, Ng Tang DS, Wang X, Kozinetz C (1999) Costimulatory pathways mediate monocyte-dependent lymphocyte apoptosis in HIV. Clin Immunol 90:302–312. doi:10.1006/clim.1998.4663 PubMedCrossRef

35.

Incardona JP, Roelink H (2000) The role of cholesterol in Shh signaling and teratogen-induced holoprosencephaly. Cell Mol Life Sci 57:1709–1719. doi:10.1007/PL00000653 PubMedCrossRef

36.

Incardona JP, Gaffield W, Lange Y et al (2000) Cyclopamine inhibition of Sonic hedgehog signal transduction is not mediated through effects on cholesterol transport. Dev Biol 224:440–452. doi:10.1006/dbio.2000.9775 PubMedCrossRef

37.

Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)). Methods 25:402–408PubMedCrossRef

38.

Chiang C, Swan RZ, Grachtchouk M et al (1999) Essential role for Sonic hedgehog during hair follicle morphogenesis. Dev Biol 205:1–9. doi:10.1006/dbio.1998.9103 PubMedCrossRef

39.

Detmer K, Walker AN, Jenkins TM, Steele TA, Dannawi H (2000) Erythroid differentiation in vitro is blocked by cyclopamine, an inhibitor of hedgehog signaling. Blood Cells Mol Dis 26:360–372. doi:10.1006/bcmd.2000.0318 PubMedCrossRef

40.

Taipale J, Chen JK, Cooper MK et al (2000) Effects of oncogenic mutations in Smoothened and Patched can be reversed by cyclopamine. Nature 406:1005–1009. doi:10.1038/35023008 PubMedCrossRef

41.

Qualtrough D, Buda A, Gaffield W, Williams AC, Paraskeva C (2004) Hedgehog signalling in colorectal tumour cells: induction of apoptosis with cyclopamine treatment. Int J Cancer 110:831–837. doi:10.1002/ijc.20227 PubMedCrossRef

42.

Zhang J, Lipinski R, Shaw A, Gipp J, Bushman W (2007) Lack of demonstrable autocrine hedgehog signaling in human prostate cancer cell lines. J Urol 177:1179–1185. doi:10.1016/j.juro.2006.10.032 PubMedCrossRef

43.

Mimeault M, Moore E, Moniaux N et al (2006) Cytotoxic effects induced by a combination of cyclopamine and gefitinib, the selective hedgehog and epidermal growth factor receptor signaling inhibitors, in prostate cancer cells. Int J Cancer 118:1022–1031. doi:10.1002/ijc.21440 PubMedCrossRef

44.

Lewis MT, Ross S, Strickland PA et al (1999) Defects in mouse mammary gland development caused by conditional haploinsufficiency of Patched-1. Development 126:5181–5193PubMed

45.

Lewis MT, Ross S, Strickland PA et al (2001) The Gli2 transcription factor is required for normal mouse mammary gland development. Dev Biol 238:133–144. doi:10.1006/dbio.2001.0410 PubMedCrossRef

Title: Cyclopamine inhibition of human breast cancer cell growth independent of Smoothened (Smo)
Authors: Xiaomei Zhang
Nikesha Harrington
Ricardo C. Moraes
Meng-Fen Wu
Susan G. Hilsenbeck
Michael T. Lewis
Publication date: 01-06-2009
Publisher: Springer US
Published in: Breast Cancer Research and Treatment / Issue 3/2009
Print ISSN: 0167-6806
Electronic ISSN: 1573-7217
DOI: https://doi.org/10.1007/s10549-008-0093-3

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 3/2009

Digital image analysis of breast epithelial cells collected by random periareolar fine-needle aspirates (RPFNA) from women at high risk for breast cancer taking hormone replacement and the aromatase inhibitor, letrozole, for six months

A pilot study of dose-dense adjuvant paclitaxel without growth factor support for women with early breast carcinoma

Oligometastatic breast cancer treated with curative-intent stereotactic body radiation therapy

DNA-repair genetic polymorphisms and risk of breast cancer in Cyprus

Autoimmune hypothyroidism and breast cancer in the elderly

Columnar cell lesions, mammographic density and breast cancer risk

Keynote webinar | Spotlight on antibody–drug conjugates in cancer