Overexpression of the cancer stem cell marker CD133 confers a poor prognosis in invasive breast cancer

  • Chitra Joseph
  • Maariya Arshad
  • Sasagu Kurozomi
  • Maryam Althobiti
  • Islam M. Miligy
  • Sara Al-izzi
  • Michael S. Toss
  • Fang Qin Goh
  • Simon J. Johnston
  • Stewart G. Martin
  • Ian O. Ellis
  • Nigel P. Mongan
  • Andrew R. Green
  • Emad A. RakhaEmail author
Preclinical study



CD133/ prominin 1 is a cancer stem cell marker associated with cancer progression and patient outcome in a variety of solid tumours, but its role in invasive breast cancer (BC) remains obscure. The current study aims to assess the prognostic value of CD133 expression in early invasive BC.


CD133 mRNA was assessed in the METABRIC cohort and at the proteomic level using immunohistochemistry utilising a large well-characterised BC cohort. Association with clinicopathological characteristics, expression of other stem cell markers and patient outcome were evaluated.


High expression of CD133 either in mRNA or protein levels was associated with characteristics of poor prognosis including high tumour grade, larger tumour size, high Nottingham Prognostic Index, HER2 positivity and hormonal receptor negativity (all; p < 0.001). High CD133 expression was positively associated with proliferation biomarkers including p16, Cyclin E and Ki67 (p < 0.01). Tumours expressing CD133 showed higher expression of other stem cell markers including CD24, CD44, SOX10, ALDHA3 and ITGA6. High expression of CD133 protein was associated with shorter BC-specific survival (p = 0.026). Multivariate analysis revealed that CD133 protein expression was an independent risk factor for shorter BC-specific survival (p = 0.038).


This study provides evidence for the prognostic value of CD133 in invasive BC. A strong positive association of BC stem cell markers is observed at the protein level. Further studies to assess the value of stem cell markers individually or in combination in BC is warranted.


Cancer Stem Cell Invasive breast cancer Prognosis CD133 



Breast cancer


BC-specific survival


Confidence intervals




Hazard ratio


Human epidermal growth factor receptor 2


Molecular taxonomy of breast cancer international consortium


Nottingham Prognostic Index




The cancer genome atlas



We thank the Nottingham Health Science Biobank and Breast Cancer Now Tissue Bank for the provision of tissue samples.

Compliance with ethical standards

Conflict of interest

The authors have no conflicts of interest to declare.

Research involving human participants

This study was approved by the Nottingham Research Ethics Committee 2 (Reference title: Development of a molecular genetic classification of breast cancer). All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Supplementary material

10549_2018_5085_MOESM1_ESM.pdf (360 kb)
Supplementary material 1 (PDF 360 KB)
10549_2018_5085_MOESM2_ESM.docx (17 kb)
Supplementary material 2 (DOCX 17 KB)
10549_2018_5085_MOESM3_ESM.docx (15 kb)
Supplementary material 3 (DOCX 15 KB)


  1. 1.
    Clarke MF, Dick JE, Dirks PB, Eaves CJ, Jamieson CH, Jones DL, Visvader J, Weissman IL, Wahl GM (2006) Cancer stem cells–perspectives on current status and future directions: AACR Workshop on cancer stem cells. Cancer research 66(19):9339–9344. Google Scholar
  2. 2.
    Dick JE (2003) Breast cancer stem cells revealed. Proc Natl Acad Sci USA 100(7):3547–3549. Google Scholar
  3. 3.
    Joseph C, Papadaki A, Althobiti M, Alsaleem M, Aleskandarany MA, Rakha EA (2018) Breast cancer intra-tumour heterogeneity: Current status and clinical implications. Histopathology. Google Scholar
  4. 4.
    Wright MH, Calcagno AM, Salcido CD, Carlson MD, Ambudkar SV, Varticovski L (2008) Brca1 breast tumours contain distinct CD44+/CD24- and CD133 + cells with cancer stem cell characteristics. Breast Cancer Res 10(1):R10. Google Scholar
  5. 5.
    Bane A, Viloria-Petit A, Pinnaduwage D, Mulligan AM, O’Malley FP, Andrulis IL (2013) Clinical-pathologic significance of cancer stem cell marker expression in familial breast cancers. Breast Cancer Res Treat 140(1):195–205. Google Scholar
  6. 6.
    Liu R, Wang X, Chen GY, Dalerba P, Gurney A, Hoey T, Sherlock G, Lewicki J, Shedden K, Clarke MF (2007) The prognostic role of a gene signature from tumourigenic breast-cancer cells. N Engl J Med 356(3):217–226. Google Scholar
  7. 7.
    Shipitsin M, Campbell LL, Argani P, Weremowicz S, Bloushtain-Qimron N, Yao J, Nikolskaya T, Serebryiskaya T, Beroukhim R, Hu M, Halushka MK, Sukumar S, Parker LM, Anderson KS, Harris LN, Garber JE, Richardson AL, Schnitt SJ, Nikolsky Y, Gelman RS, Polyak K (2007) Molecular definition of breast tumour heterogeneity. Cancer Cell 11(3):259–273. Google Scholar
  8. 8.
    Horst D, Kriegl L, Engel J, Kirchner T, Jung A (2008) CD133 expression is an independent prognostic marker for low survival in colorectal cancer. Br J Cancer 99(8):1285–1289Google Scholar
  9. 9.
    Madjd Z, Mehrjerdi AZ, Sharifi AM, Molanaei S, Shahzadi SZ, Asadi-Lari M (2009) CD44 + cancer cells express higher levels of the anti-apoptotic protein Bcl-2 in breast tumours. Cancer Immun 9:4Google Scholar
  10. 10.
    Liu G, Yuan X, Zeng Z, Tunici P, Ng H, Abdulkadir IR, Lu L, Irvin D, Black KL, Yu JS (2006) Analysis of gene expression and chemoresistance of CD133 + cancer stem cells in glioblastoma. Mol Cancer 5:67. Google Scholar
  11. 11.
    Tabu K, Kimura T, Sasai K, Wang L, Bizen N, Nishihara H, Taga T, Tanaka S (2010) Analysis of an alternative human CD133 promoter reveals the implication of Ras/ERK pathway in tumour stem-like hallmarks. Mol Cancer 9:39. Google Scholar
  12. 12.
    Wang YK, Zhu YL, Qiu FM, Zhang T, Chen ZG, Zheng S, Huang J (2010) Activation of Akt and MAPK pathways enhances the tumourigenicity of CD133 + primary colon cancer cells. Carcinogenesis 31(8):1376–1380. Google Scholar
  13. 13.
    Brugnoli F, Grassilli S, Piazzi M, Palomba M, Nika E, Bavelloni A, Capitani S, Bertagnolo V (2013) In triple negative breast tumour cells, PLC-beta2 promotes the conversion of CD133high to CD133low phenotype and reduces the CD133-related invasiveness. Mol Cancer 12:165. Google Scholar
  14. 14.
    Zhao P, Lu Y, Jiang X, Li X (2011) Clinicopathological significance and prognostic value of CD133 expression in triple-negative breast carcinoma. Cancer Sci 102(5):1107–1111. Google Scholar
  15. 15.
    Zhu Y, Kong F, Zhang C, Ma C, Xia H, Quan B, Cui H (2017) CD133 mediates the TGF-beta1-induced activation of the PI3K/ERK/P70S6K signaling pathway in gastric cancer cells. Oncol Lett 14(6):7211–7216. Google Scholar
  16. 16.
    Shao GL, Wang MC, Fan XL, Zhong L, Ji SF, Sang G, Wang S (2018) Correlation between Raf/MEK/ERK signaling pathway and clinicopathological features and prognosis for patients with breast cancer having axillary lymph node metastasis. Technol Cancer Res Treat 17:1533034617754024. Google Scholar
  17. 17.
    Abd El-Rehim DM, Ball G, Pinder SE, Rakha E, Paish C, Robertson JF, Macmillan D, Blamey RW, Ellis IO (2005) High-throughput protein expression analysis using tissue microarray technology of a large well-characterised series identifies biologically distinct classes of breast cancer confirming recent cDNA expression analyses. Int J Cancer 116(3):340–350. Google Scholar
  18. 18.
    Aleskandarany MA, Green AR, Benhasouna AA, Barros FF, Neal K, Reis-Filho JS, Ellis IO, Rakha EA (2012) Prognostic value of proliferation assay in the luminal, HER2-positive, and triple-negative biologic classes of breast cancer. Breast Cancer Res 14(1):R3. Google Scholar
  19. 19.
    Rakha EA, Elsheikh SE, Aleskandarany MA, Habashi HO, Green AR, Powe DG, El-Sayed ME, Benhasouna A, Brunet JS, Akslen LA, Evans AJ, Blamey R, Reis-Filho JS, Foulkes WD, Ellis IO (2009) Triple-negative breast cancer: distinguishing between basal and nonbasal subtypes. Clin Cancer Res 15(7):2302–2310. Google Scholar
  20. 20.
    Curtis C, Shah SP, Chin SF, Turashvili G, Rueda OM, Dunning MJ, Speed D, Lynch AG, Samarajiwa S, Yuan Y, Graf S, Ha G, Haffari G, Bashashati A, Russell R, McKinney S, Group M, Langerod A, Green A, Provenzano E, Wishart G, Pinder S, Watson P, Markowetz F, Murphy L, Ellis I, Purushotham A, Borresen-Dale AL, Brenton JD, Tavare S, Caldas C, Aparicio S (2012) The genomic and transcriptomic architecture of 2,000 breast tumours reveals novel subgroups. Nature 486(7403):346–352. Google Scholar
  21. 21.
    Pereira B, Chin SF, Rueda OM, Vollan HK, Provenzano E, Bardwell HA, Pugh M, Jones L, Russell R, Sammut SJ, Tsui DW, Liu B, Dawson SJ, Abraham J, Northen H, Peden JF, Mukherjee A, Turashvili G, Green AR, McKinney S, Oloumi A, Shah S, Rosenfeld N, Murphy L, Bentley DR, Ellis IO, Purushotham A, Pinder SE, Borresen-Dale AL, Earl HM, Pharoah PD, Ross MT, Aparicio S, Caldas C (2016) The somatic mutation profiles of 2,433 breast cancers refines their genomic and transcriptomic landscapes. Nat Commun 7:11479. Google Scholar
  22. 22.
    Jezequel P, Campone M, Gouraud W, Guerin-Charbonnel C, Leux C, Ricolleau G, Campion L (2012) bc-GenExMiner: an easy-to-use online platform for gene prognostic analyses in breast cancer. Breast Cancer Res Treat 131(3):765–775. Google Scholar
  23. 23.
    Joseph C, Macnamara O, Craze M, Russell R, Provenzano E, Nolan CC, Diez-Rodriguez M, Sonbul SN, Aleskandarany MA, Green AR, Rakha EA, Ellis IO, Mukherjee A (2018) Mediator complex (MED) 7: a biomarker associated with good prognosis in invasive breast cancer, especially ER+ luminal subtypes. Br J Cancer 118(8):1142–1151. Google Scholar
  24. 24.
    Kurozumi S, Joseph C, Sonbul S, Gorringe KL, Pigera M, Aleskandarany MA, Diez-Rodriguez M, Nolan CC, Fujii T, Shirabe K, Kuwano H, Storr S, Martin SG, Ellis IO, Green AR, Rakha EA (2018) Clinical and biological roles of Kelch-like family member 7 in breast cancer: a marker of poor prognosis. Breast Cancer Res Treat. Google Scholar
  25. 25.
    Cancer Genome Atlas N (2012) Comprehensive molecular portraits of human breast tumours. Nature 490(7418):61–70. Google Scholar
  26. 26.
    McCarty KS Jr, McCarty KS, Sr (1984) Histochemical approaches to steroid receptor analyses. Semin Diagn Pathol 1(4):297–308Google Scholar
  27. 27.
    Aleskandarany MA, Rakha EA, Ahmed MA, Powe DG, Paish EC, Macmillan RD, Ellis IO, Green AR (2010) PIK3CA expression in invasive breast cancer: a biomarker of poor prognosis. Breast Cancer Res Treat 122(1):45–53. Google Scholar
  28. 28.
    Aleskandarany MA, Rakha EA, Macmillan RD, Powe DG, Ellis IO, Green AR (2011) MIB1/Ki-67 labelling index can classify grade 2 breast cancer into two clinically distinct subgroups. Breast Cancer Res Treat 127(3):591–599. Google Scholar
  29. 29.
    Alshareeda AT, Soria D, Garibaldi JM, Rakha E, Nolan C, Ellis IO, Green AR (2013) Characteristics of basal cytokeratin expression in breast cancer. Breast Cancer Res Treat 139(1):23–37. Google Scholar
  30. 30.
    Li X, Zhao H, Gu J, Zheng L (2015) Prognostic value of cancer stem cell marker CD133 expression in pancreatic ductal adenocarcinoma (PDAC): a systematic review and meta-analysis. Int J Clin Exp Pathol 8(10):12084–12092Google Scholar
  31. 31.
    Filippini SE, Vega A (2013) Breast cancer genes: beyond BRCA1 and BRCA2. Front Biosci 18:1358–1372Google Scholar
  32. 32.
    Horst D, Kriegl L, Engel J, Kirchner T, Jung A (2009) Prognostic significance of the cancer stem cell markers CD133, CD44, and CD166 in colorectal cancer. Cancer Investig 27(8):844–850. Google Scholar
  33. 33.
    Silva IA, Bai S, McLean K, Yang K, Griffith K, Thomas D, Ginestier C, Johnston C, Kueck A, Reynolds RK, Wicha MS, Buckanovich RJ (2011) Aldehyde dehydrogenase in combination with CD133 defines angiogenic ovarian cancer stem cells that portend poor patient survival. Cancer Res 71(11):3991–4001. Google Scholar
  34. 34.
    Zeppernick F, Ahmadi R, Campos B, Dictus C, Helmke BM, Becker N, Lichter P, Unterberg A, Radlwimmer B, Herold-Mende CC (2008) Stem cell marker CD133 affects clinical outcome in glioma patients. Clinical Cancer Res 14(1):123–129. Google Scholar
  35. 35.
    Kim SJ, Kim YS, Jang ED, Seo KJ, Kim JS (2015) Prognostic impact and clinicopathological correlation of CD133 and ALDH1 expression in invasive breast cancer. J Breast Cancer 18(4):347–355. Google Scholar
  36. 36.
    Zhang J, Guo X, Chang DY, Rosen DG, Mercado-Uribe I, Liu J (2012) CD133 expression associated with poor prognosis in ovarian cancer. Mod Pathol 25(3):456–464. Google Scholar
  37. 37.
    Hashimoto K, Aoyagi K, Isobe T, Kouhuji K, Shirouzu K (2014) Expression of CD133 in the cytoplasm is associated with cancer progression and poor prognosis in gastric cancer. Gastric Cancer 17(1):97–106. Google Scholar
  38. 38.
    Saeednejad Zanjani L, Madjd Z, Abolhasani M, Andersson Y, Rasti A, Shariftabrizi A, Asgari M (2017) Cytoplasmic expression of CD133 stemness marker is associated with tumour aggressiveness in clear cell renal cell carcinoma. Exp Mol Pathol 103(2):218–228. Google Scholar
  39. 39.
    Xia P (2017) CD133 mRNA may be a suitable prognostic marker for human breast cancer. Stem Cell Investig 4:87. Google Scholar
  40. 40.
    Gluz O, Liedtke C, Gottschalk N, Pusztai L, Nitz U, Harbeck N (2009) Triple-negative breast cancer–current status and future directions. Ann Oncol 20(12):1913–1927. Google Scholar
  41. 41.
    Bertagnolo V, Benedusi M, Querzoli P, Pedriali M, Magri E, Brugnoli F, Capitani S (2006) PLC-beta2 is highly expressed in breast cancer and is associated with a poor outcome: a study on tissue microarrays. Int J Oncol 28(4):863–872Google Scholar
  42. 42.
    Brugnoli F, Grassilli S, Lanuti P, Marchisio M, Al-Qassab Y, Vezzali F, Capitani S, Bertagnolo V (2017) Up-modulation of PLC-beta2 reduces the number and malignancy of triple-negative breast tumour cells with a CD133(+)/EpCAM(+) phenotype: a promising target for preventing progression of TNBC. BMC Cancer 17(1):617. Google Scholar
  43. 43.
    Dublin EA, Patel NK, Gillett CE, Smith P, Peters G, Barnes DM (1998) Retinoblastoma and p16 proteins in mammary carcinoma: their relationship to cyclin D1 and histopathological parameters. Int J Cancer 79(1):71–75Google Scholar
  44. 44.
    Sher CJ (1996) Cancer cell cycle. Science 274:1672–1677Google Scholar
  45. 45.
    Casimiro MC, Crosariol M, Loro E, Li Z, Pestell RG (2012) Cyclins and cell cycle control in cancer and disease. Genes Cancer 3(11–12):649–657. Google Scholar
  46. 46.
    Hemmings BA, Restuccia DF (2015) The PI3K-PKB/Akt pathway. Cold Spring Harbor Perspect Biol 7 (4).
  47. 47.
    Li L, Xie T (2005) Stem cell niche: structure and function. Ann Rev Cell Dev Biol 21:605–631. Google Scholar
  48. 48.
    Paling NR, Wheadon H, Bone HK, Welham MJ (2004) Regulation of embryonic stem cell self-renewal by phosphoinositide 3-kinase-dependent signaling. J Biol Chem 279(46):48063–48070. Google Scholar
  49. 49.
    Young CD, Zimmerman LJ, Hoshino D, Formisano L, Hanker AB, Gatza ML, Morrison MM, Moore PD, Whitwell CA, Dave B, Stricker T, Bhola NE, Silva GO, Patel P, Brantley-Sieders DM, Levin M, Horiates M, Palma NA, Wang K, Stephens PJ, Perou CM, Weaver AM, O’Shaughnessy JA, Chang JC, Park BH, Liebler DC, Cook RS, Arteaga CL (2015) Activating PIK3CA Mutations Induce an Epidermal Growth Factor Receptor (EGFR)/Extracellular Signal-regulated Kinase (ERK) Paracrine Signaling Axis in Basal-like Breast Cancer. Mol Cell Proteomics 14(7):1959–1976. Google Scholar
  50. 50.
    Bourguignon LY, Peyrollier K, Xia W, Gilad E (2008) Hyaluronan-CD44 interaction activates stem cell marker Nanog, Stat-3-mediated MDR1 gene expression, and ankyrin-regulated multidrug efflux in breast and ovarian tumour cells. J Biol Chem 283(25):17635–17651. Google Scholar
  51. 51.
    de la Torre M, Heldin P, Bergh J (1995) Expression of the CD44 glycoprotein (lymphocyte-homing receptor) in untreated human breast cancer and its relationship to prognostic markers. Anticancer Res 15(6B):2791–2795Google Scholar
  52. 52.
    Shakhova O, Zingg D, Schaefer SM, Hari L, Civenni G, Blunschi J, Claudinot S, Okoniewski M, Beermann F, Mihic-Probst D, Moch H, Wegner M, Dummer R, Barrandon Y, Cinelli P, Sommer L (2012) Sox10 promotes the formation and maintenance of giant congenital naevi and melanoma. Nature Cell Biol 14(8):882–890. Google Scholar
  53. 53.
    Moon SH, Kim DK, Cha Y, Jeon I, Song J, Park KS (2013) PI3K/Akt and Stat3 signaling regulated by PTEN control of the cancer stem cell population, proliferation and senescence in a glioblastoma cell line. Int J Oncol 42(3):921–928. Google Scholar
  54. 54.
    Stambolic V, Suzuki A, de la Pompa JL, Brothers GM, Mirtsos C, Sasaki T, Ruland J, Penninger JM, Siderovski DP, Mak TW (1998) Negative regulation of PKB/Akt-dependent cell survival by the tumour suppressor PTEN. Cell 95(1):29–39Google Scholar
  55. 55.
    Liu Q, Li JG, Zheng XY, Jin F, Dong HT (2009) Expression of CD133, PAX2, ESA, and GPR30 in invasive ductal breast carcinomas. Chin Med J 122(22):2763–2769Google Scholar
  56. 56.
    Schmohl JU, Vallera DA (2016) CD133, selectively targeting the root of cancer. Toxins 8(6):165. Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Chitra Joseph
    • 1
  • Maariya Arshad
    • 1
  • Sasagu Kurozomi
    • 1
    • 2
  • Maryam Althobiti
    • 1
  • Islam M. Miligy
    • 1
    • 5
  • Sara Al-izzi
    • 1
  • Michael S. Toss
    • 1
    • 5
  • Fang Qin Goh
    • 1
  • Simon J. Johnston
    • 1
  • Stewart G. Martin
    • 1
  • Ian O. Ellis
    • 1
    • 6
  • Nigel P. Mongan
    • 3
    • 4
  • Andrew R. Green
    • 1
  • Emad A. Rakha
    • 1
    • 5
    • 6
    Email author
  1. 1.Nottingham Breast Cancer Research Centre, Division of Cancer and Stem Cells, School of MedicineUniversity of NottinghamNottinghamUK
  2. 2.Department of General Surgical ScienceGunma University Graduate School of MedicineGunmaJapan
  3. 3.Cancer Biology and Translational Research, Faculty of Medicine and Health SciencesUniversity of NottinghamNottinghamUK
  4. 4.Department of PharmacologyWeill Cornell MedicineNew YorkUSA
  5. 5.Histopathology Department, Faculty of MedicineMenoufia UniversityShebin El-komEgypt
  6. 6.Department of Histopathology, School of MedicineNottingham University Hospitals NHS Trust, Nottingham City HospitalNottinghamUK

Personalised recommendations