Advertisement

Breast Cancer Research and Treatment

, Volume 158, Issue 1, pp 29–41 | Cite as

Transcriptomic profiling of curcumin-treated human breast stem cells identifies a role for stearoyl-coa desaturase in breast cancer prevention

  • Justin A. Colacino
  • Sean P. McDermott
  • Maureen A. Sartor
  • Max S. Wicha
  • Laura S. Rozek
Preclinical study

Abstract

Curcumin is a potential agent for both the prevention and treatment of cancers. Curcumin treatment alone, or in combination with piperine, limits breast stem cell self-renewal, while remaining non-toxic to normal differentiated cells. We paired fluorescence-activated cell sorting with RNA sequencing to characterize the genome-wide changes induced specifically in normal breast stem cells following treatment with these compounds. We generated genome-wide maps of the transcriptional changes that occur in epithelial-like (ALDH+) and mesenchymal-like (ALDH−/CD44+/CD24−) normal breast stem/progenitor cells following treatment with curcumin and piperine. We show that curcumin targets both stem cell populations by down-regulating expression of breast stem cell genes including ALDH1A3, CD49f, PROM1, and TP63. We also identified novel genes and pathways targeted by curcumin, including downregulation of SCD. Transient siRNA knockdown of SCD in MCF10A cells significantly inhibited mammosphere formation and the mean proportion of CD44+/CD24− cells, suggesting that SCD is a regulator of breast stemness and a target of curcumin in breast stem cells. These findings extend previous reports of curcumin targeting stem cells, here in two phenotypically distinct stem/progenitor populations isolated from normal human breast tissue. We identified novel mechanisms by which curcumin and piperine target breast stem cell self-renewal, such as by targeting lipid metabolism, providing a mechanistic link between curcumin treatment and stem cell self-renewal. These results elucidate the mechanisms by which curcumin may act as a cancer-preventive compound and provide novel targets for cancer prevention and treatment.

Keywords

Curcumin Breast stem cell RNA-seq Stearoyl-coa desaturase Prevention 

Notes

Acknowledgments

Support for this study was provided by a grant from the National Cancer Institute (R03 CA167700). Support for JAC was provided by the Rackham Predoctoral Fellowship from the University of Michigan and Institutional Training Grants from the National Institute of Environmental Health Sciences (NIEHS) (T32 ES007062) and the National Human Genome Research Institute (NHGRI) (T32 HG00040).

Compliance with ethical standards

Conflict of interest

The authors have no conflicts of interest to declare.

Supplementary material

10549_2016_3854_MOESM1_ESM.xlsx (291 kb)
Supplementary material 1 (XLSX 290 kb)
10549_2016_3854_MOESM2_ESM.docx (591 kb)
Supplementary material 2 (DOCX 591 kb)

References

  1. 1.
    Jemal A et al (2010) Cancer statistics, 2010. CA Cancer J Clin 60(5):277–300CrossRefPubMedGoogle Scholar
  2. 2.
    Brandberg Y et al (2008) Psychological reactions, quality of life, and body image after bilateral prophylactic mastectomy in women at high risk for breast cancer: a prospective 1-year follow-up study. J Clin Oncol 26(24):3943–3949CrossRefPubMedGoogle Scholar
  3. 3.
    Fisher B et al (1998) Tamoxifen for prevention of breast cancer: report of the National Surgical Adjuvant Breast and Bowel Project P-1 Study. J Natl Cancer Inst 90(18):1371–1388CrossRefPubMedGoogle Scholar
  4. 4.
    Vogel VG et al (2010) Update of the national surgical adjuvant breast and bowel project study of tamoxifen and raloxifene (STAR) P-2 Trial: preventing breast cancer. Cancer Prev Res (Phila) 3(6):696–706CrossRefGoogle Scholar
  5. 5.
    Howell A (2008) The endocrine prevention of breast cancer. Best Pract Res Clin Endocrinol Metab 22(4):615–623CrossRefPubMedGoogle Scholar
  6. 6.
    Noorafshan A, Ashkani-Esfahani S (2013) A review of therapeutic effects of curcumin. Curr Pharm Des 19(11):2032–2046PubMedGoogle Scholar
  7. 7.
    Shoba G et al (1998) Influence of piperine on the pharmacokinetics of curcumin in animals and human volunteers. Planta Med 64(4):353–356CrossRefPubMedGoogle Scholar
  8. 8.
    Dontu G et al (2003) In vitro propagation and transcriptional profiling of human mammary stem/progenitor cells. Genes Dev 17(10):1253–1270CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Kakarala M et al (2010) Targeting breast stem cells with the cancer preventive compounds curcumin and piperine. Breast Cancer Res Treat 122(3):777–785CrossRefPubMedGoogle Scholar
  10. 10.
    Molyneux G et al (2010) BRCA1 basal-like breast cancers originate from luminal epithelial progenitors and not from basal stem cells. Cell Stem Cell 7(3):403–417CrossRefPubMedGoogle Scholar
  11. 11.
    Al-Hajj M et al (2003) Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci USA 100(7):3983–3988CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Van Keymeulen A et al (2011) Distinct stem cells contribute to mammary gland development and maintenance. Nature 479(7372):189–193CrossRefPubMedGoogle Scholar
  13. 13.
    Liu S et al (2013) Breast cancer stem cells transition between epithelial and mesenchymal states reflective of their normal counterparts. Stem Cell Reports 2(1):78–91CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Dobin A et al (2013) STAR: ultrafast universal RNA-seq aligner. Bioinformatics 29(1):15–21CrossRefPubMedGoogle Scholar
  15. 15.
    Robinson MD, McCarthy DJ, Smyth GK (2010) edgeR: a bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 26(1):139–140CrossRefPubMedGoogle Scholar
  16. 16.
    Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Stat Soc Ser B (Methodological), p 289–300Google Scholar
  17. 17.
    Lim E et al (2009) Aberrant luminal progenitors as the candidate target population for basal tumor development in BRCA1 mutation carriers. Nat Med 15(8):907–913CrossRefPubMedGoogle Scholar
  18. 18.
    Svedberg J et al (1990) Free-fatty acid inhibition of insulin binding, degradation, and action in isolated rat hepatocytes. Diabetes 39(5):570–574CrossRefPubMedGoogle Scholar
  19. 19.
    Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25(4):402–408CrossRefPubMedGoogle Scholar
  20. 20.
    R Core Team R: A language and environment for statistical computing. 2013: R Foundation for Statistical Computing, Vienna, AustriaGoogle Scholar
  21. 21.
    Eirew P et al (2012) Aldehyde dehydrogenase activity is a biomarker of primitive normal human mammary luminal cells. Stem Cells 30(2):344–348CrossRefPubMedGoogle Scholar
  22. 22.
    Keller PJ et al (2012) Defining the cellular precursors to human breast cancer. Proc Natl Acad Sci USA 109(8):2772–2777CrossRefPubMedGoogle Scholar
  23. 23.
    Isfoss BL et al (2013) Women with familial risk for breast cancer have an increased frequency of aldehyde dehydrogenase expressing cells in breast ductules. BMC Clin Pathol 13(1):28CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Kunju LP et al (2011) EZH2 and ALDH-1 mark breast epithelium at risk for breast cancer development. Mod Pathol 24(6):786–793CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Zhong Y et al (2013) Expression of ALDH1 in breast invasive ductal carcinoma: an independent predictor of early tumor relapse. Cancer Cell Int 13(1):60CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Sheridan C et al (2006) CD44+/CD24− breast cancer cells exhibit enhanced invasive properties: an early step necessary for metastasis. Breast Cancer Res 8(5):R59CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Mani SA et al (2008) The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell 133(4):704–715CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Yang C et al (2009) Curcumin upregulates transcription factor Nrf2, HO-1 expression and protects rat brains against focal ischemia. Brain Res 1282:133–141CrossRefPubMedGoogle Scholar
  29. 29.
    McNally SJ et al (2007) Curcumin induces heme oxygenase 1 through generation of reactive oxygen species, p38 activation and phosphatase inhibition. Int J Mol Med 19(1):165–172PubMedGoogle Scholar
  30. 30.
    Motterlini R et al (2000) Curcumin, an antioxidant and anti-inflammatory agent, induces heme oxygenase-1 and protects endothelial cells against oxidative stress. Free Radic Biol Med 28(8):1303–1312CrossRefPubMedGoogle Scholar
  31. 31.
    Shen SQ et al (2007) Protective effect of curcumin against liver warm ischemia/reperfusion injury in rat model is associated with regulation of heat shock protein and antioxidant enzymes. World J Gastroenterol 13(13):1953–1961CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Dunsmore KE, Chen PG, Wong HR (2001) Curcumin, a medicinal herbal compound capable of inducing the heat shock response. Crit Care Med 29(11):2199–2204CrossRefPubMedGoogle Scholar
  33. 33.
    Newman B et al (2012) HSP90 inhibitor 17-AAG selectively eradicates lymphoma stem cells. Cancer Res 72(17):4551–4561CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Lee CH et al (2012) Inhibition of heat shock protein (Hsp) 27 potentiates the suppressive effect of Hsp90 inhibitors in targeting breast cancer stem-like cells. Biochimie 94(6):1382–1389CrossRefPubMedGoogle Scholar
  35. 35.
    Subramaniam D et al (2012) Curcumin induces cell death in esophageal cancer cells through modulating Notch signaling. PLoS One 7(2):e30590CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Charpentier MS et al (2014) Curcumin targets breast cancer stem-like cells with microtentacles that persist in mammospheres and promote reattachment. Cancer Res 74(4):1250–1260CrossRefPubMedGoogle Scholar
  37. 37.
    Whipple RA et al (2008) Vimentin filaments support extension of tubulin-based microtentacles in detached breast tumor cells. Cancer Res 68(14):5678–5688CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Bhardwaj RK et al (2002) Piperine, a major constituent of black pepper, inhibits human P-glycoprotein and CYP3A4. J Pharmacol Exp Ther 302(2):645–650CrossRefPubMedGoogle Scholar
  39. 39.
    Duangjai A et al (2013) Black pepper and piperine reduce cholesterol uptake and enhance translocation of cholesterol transporter proteins. J Nat Med 67(2):303–310CrossRefPubMedGoogle Scholar
  40. 40.
    Hong J et al (2004) Modulation of arachidonic acid metabolism by curcumin and related β-diketone derivatives: effects on cytosolic phospholipase A2, cyclooxygenases and 5-lipoxygenase. Carcinogenesis 25(9):1671–1679CrossRefPubMedGoogle Scholar
  41. 41.
    Lev-Ari S et al (2006) Down-regulation of prostaglandin E2 by curcumin is correlated with inhibition of cell growth and induction of apoptosis in human colon carcinoma cell lines. J Soc Integr Oncol 4(1):21–26PubMedGoogle Scholar
  42. 42.
    Kudo C et al (2011) Novel curcumin analogs, GO-Y030 and GO-Y078, are multi-targeted agents with enhanced abilities for multiple myeloma. Anticancer Res 31(11):3719–3726PubMedGoogle Scholar
  43. 43.
    Shin HS et al (2014) Anti-atherosclerosis and hyperlipidemia effects of herbal mixture, Artemisia iwayomogi Kitamura and Curcuma longa Linne, in apolipoprotein E-deficient mice. J Ethnopharmacol 153(1):142–150CrossRefPubMedGoogle Scholar
  44. 44.
    Igal RA (2010) Stearoyl-CoA desaturase-1: a novel key player in the mechanisms of cell proliferation, programmed cell death and transformation to cancer. Carcinogenesis 31(9):1509–1515CrossRefPubMedGoogle Scholar
  45. 45.
    Noto A et al (2013) Stearoyl-CoA desaturase-1 is a key factor for lung cancer-initiating cells. Cell Death Dis 4:e947CrossRefPubMedPubMedCentralGoogle Scholar
  46. 46.
    Mauvoisin D et al (2013) Decreasing stearoyl-CoA desaturase-1 expression inhibits beta-catenin signaling in breast cancer cells. Cancer Sci 104(1):36–42CrossRefPubMedGoogle Scholar
  47. 47.
    Rios-Esteves J, Marilyn Resh D (2013) Stearoyl CoA desaturase is required to produce active, lipid-modified Wnt proteins. Cell Rep 4(6):1072–1081CrossRefPubMedGoogle Scholar
  48. 48.
    Coleman DT et al (2015) Curcumin prevents palmitoylation of integrin beta4 in breast cancer cells. PLoS One 10(5):e0125399CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of Environmental Health SciencesUniversity of Michigan School of Public HealthAnn ArborUSA
  2. 2.University of Michigan Comprehensive Cancer CenterAnn ArborUSA
  3. 3.Department of Nutritional SciencesUniversity of Michigan School of Public HealthAnn ArborUSA
  4. 4.Department of Computational Medicine and BioinformaticsUniversity of MichiganAnn ArborUSA

Personalised recommendations