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Cited by 27Wnt/β-catenin signaling in development and cancerCancer Cells and MetastasisEstrogen and related hormone effects

Human Breast Progenitor Cell Numbers Are Regulated by WNT and TBX3

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Arendt, Lisa M., St. Laurent, Jessica, Wronski, Ania, Caballero, Silvia, Lyle, Stephen R., Naber, Stephen P., Kuperwasser, Charlotte·2014·PLoS ONE
DOI 10.1371/journal.pone.0111442PMC PMC4211891
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Abstract

BACKGROUND: Although human breast development is mediated by hormonal and non-hormonal means, the mechanisms that regulate breast progenitor cell activity remain to be clarified. This limited understanding of breast progenitor cells has been due in part to the lack of appropriate model systems to detect and characterize their properties. METHODS: To examine the effects of WNT signaling and TBX3 expression on progenitor activity in the breast, primary human mammary epithelial cells (MEC) were isolated from reduction mammoplasty tissues and transduced with lentivirus to overexpress WNT1 or TBX3 or reduce expression of their cognate receptors using shRNA. Changes in progenitor activity were quantified using characterized assays. We identified WNT family members expressed by cell populations within the epithelium and assessed alterations in expression of WNT family ligands by MECs in response to TBX3 overexpression and treatment with estrogen and progesterone. RESULTS: Growth of MECs on collagen gels resulted in the formation of distinct luminal acinar and basal ductal colonies. Overexpression of TBX3 in MECs resulted in increased ductal colonies, while shTBX3 expression diminished both colony types. Increased WNT1 expression led to enhanced acinar colony formation, shLRP6 decreased both types of colonies. Estrogen stimulated the formation of acinar colonies in control MEC, but not shLRP6 MEC. Formation of ductal colonies was enhanced in response to progesterone. However, while shLRP6 decreased MEC responsiveness to progesterone, shTBX3 expression did not alter this response. CONCLUSIONS: We identified two phenotypically distinguishable lineage-committed progenitor cells that contribute to different structural elements and are regulated via hormonal and non-hormonal mechanisms. WNT signaling regulates both types of progenitor activity. Progesterone favors the expansion of ductal progenitor cells, while estrogen stimulates the expansion of acinar progenitor cells. Paracrine WNT signaling is stimulated by estrogen and progesterone, while autocrine WNT signaling is induced by the embryonic T-box transcription factor TBX3.

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  • Tbx3 is involved in the carcinogenesis of multiple types of human cancers.

    “As Tbx3 is involved in the carcinogenesis of multiple types of human cancers, our findings suggest an important target for anti-cancer drug development.”

    From Novel HDAC5-interacting motifs of Tbx3 are essential for the suppression of E-cadherin expression and for the promotion of metastasis in hepatocellular carcinoma (2018) · cites this paper as [23]

    The Discussion (page unavailable) says TBX3 mutations in breast cancer suggest a possible role in its genesis and reports increased TBX3 activity in various cancer cell lines. It does not establish TBX3 involvement in carcinogenesis across multiple types of human cancer; it says further study is needed even for its role in breast tumors. (LLM, conf=0.85)

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Research works printed but not linked: 56

  1. [2]Keller PJ, Arendt LM, Skibinski A, Logvinenko T, Klebba I, et al (2012) Defining the cellular precursors to human breast cancer. Proc Natl Acad Sci U S A. 109: 2772– 2777. PMC3286919 21940501
  2. [3]Spike BT, Engle DD, Lin JC, Cheung SK, La J, et al (2012) A mammary stem cell population identified and characterized in late embryogenesis reveals similarities to human breast cancer. Cell Stem Cell 10: 183– 197 S1934-5909(12)00002-1 [pii]; 10.1016/j.stem.2011.12.018 [doi] 22305568 PMC3277444 · DOI
  3. [4]van Amerongen R, Bowman AN, Nusse R (2012) Developmental stage and time dictate the fate of Wnt/beta-catenin-responsive stem cells in the mammary gland. Cell Stem Cell 11: 387– 400 S1934-5909(12)00342-6 [pii]; 10.1016/j.stem.2012.05.023 [doi] 22863533 PMC13155203 · DOI
  4. [5]Rios AC, Fu NY, Lindeman GJ, Visvader JE (2014) In situ identification of bipotent stem cells in the mammary gland. Nature 506: 322– 327 nature12948 [pii]; 10.1038/nature12948 [doi] 24463516 · DOI
  5. [6]Brisken C (2013) Progesterone signalling in breast cancer: a neglected hormone coming into the limelight. Nat Rev Cancer 13: 385– 396 nrc3518 [pii]; 10.1038/nrc3518 [doi] 23702927 · DOI
  6. [7]Hovey RC, Trott JF, Vonderhaar BK (2002) Establishing a framework for the functional mammary gland: from endocrinology to morphology. J Mammary Gland Biol Neoplasia 7: 17– 38. 12160083
  7. [8]Roarty K, Rosen JM (2010) Wnt and mammary stem cells: hormones cannot fly wingless. Curr Opin Pharmacol 10: 643– 649 S1471-4892(10)00125-6 [pii]; 10.1016/j.coph.2010.07.004 [doi] 20810315 PMC2981611 · DOI
  8. [9]Oakes SR, Rogers RL, Naylor MJ, Ormandy CJ (2008) Prolactin regulation of mammary gland development. J Mammary Gland Biol Neoplasia 13: 13– 28. 18219564
  9. [10]Brisken C, Park S, Vass T, Lydon JP, O'Malley BW, et al (1998) A paracrine role for the epithelial progesterone receptor in mammary gland development. Proc Natl Acad Sci U S A 95: 5076– 5081. 9560231 PMC20216
  10. [11]Mallepell S, Krust A, Chambon P, Brisken C (2006) Paracrine signaling through the epithelial estrogen receptor alpha is required for proliferation and morphogenesis in the mammary gland. Proc Natl Acad Sci U S A 103: 2196– 2201. 16452162 PMC1413744
  11. [12]Bocchinfuso WP, Korach KS (1997) Mammary gland development and tumorigenesis in estrogen receptor knockout mice. J Mammary Gland Biol Neoplasia 2: 323– 334. 10935020
  12. [13]Ciarloni L, Mallepell S, Brisken C (2007) Amphiregulin is an essential mediator of estrogen receptor alpha function in mammary gland development. Proc Natl Acad Sci U S A 104: 5455– 5460. 17369357 PMC1838509
  13. [14]Kenney NJ, Bowman A, Korach KS, Barrett JC, Salomon DS (2003) Effect of exogenous epidermal-like growth factors on mammary gland development and differentiation in the estrogen receptor-alpha knockout (ERKO) mouse. Breast Cancer Res Treat 79: 161– 173. 12825851
  14. [15]Raafat AM, Li S, Bennett JM, Hofseth LJ, Haslam SZ (2001) Estrogen and estrogen plus progestin act directly on the mammary gland to increase proliferation in a postmenopausal mouse model. J Cell Physiol 187: 81– 89 10.1002/1097-4652(2001)9999:9999<::AID-JCP1056>3.0.CO;2-0 [pii];10.1002/1097-4652(2001)9999:9999<::AID-JCP1056>3.0.CO;2-0 [doi] 11241352 · DOI
  15. [16]Jones FE, Jerry DJ, Guarino BC, Andrews GC, Stern DF (1996) Heregulin induces in vivo proliferation and differentiation of mammary epithelium into secretory lobuloalveoli. Cell Growth Differ 7: 1031– 1038. 8853899
  16. [17]Tanos T, Sflomos G, Echeverria PC, Ayyanan A, Gutierrez M, et al (2013) Progesterone/RANKL is a major regulatory axis in the human breast. Sci Transl Med 5: 182ra55 5/182/182ra55 [pii]; 10.1126/scitranslmed.3005654 [doi] 23616122 · DOI
  17. [18]Chapman DL, Garvey N, Hancock S, Alexiou M, Agulnik SI, et al (1996) Expression of the T-box family genes, Tbx1–Tbx5, during early mouse development. Dev Dyn 206: 379– 390 10.1002/(SICI)1097-0177(199608)206: 4<379::AID-AJA4>3.0.CO;2-F [pii];10.1002/(SICI)1097-0177(199608)206: 4<379::AID-AJA4>3.0.CO;2-F [doi] 8853987 · DOI
  18. [19]Eblaghie MC, Song SJ, Kim JY, Akita K, Tickle C, et al (2004) Interactions between FGF and Wnt signals and Tbx3 gene expression in mammary gland initiation in mouse embryos. J Anat 205: 1– 13. 15255957 PMC1571327
  19. [20]Davenport TG, Jerome-Majewska LA, Papaioannou VE (2003) Mammary gland, limb and yolk sac defects in mice lacking Tbx3, the gene mutated in human ulnar mammary syndrome. Development 130: 2263– 2273. 12668638
  20. [21]Bamshad M, Le T, Watkins WS, Dixon ME, Kramer BE, et al (1999) The spectrum of mutations in TBX3: Genotype/Phenotype relationship in ulnar-mammary syndrome. Am J Hum Genet 64: 1550– 1562. 10330342 PMC1377898
  21. [22]Bamshad M, Lin RC, Law DJ, Watkins WC, Krakowiak PA, et al (1997) Mutations in human TBX3 alter limb, apocrine and genital development in ulnar-mammary syndrome. Nat Genet 16: 311– 315. 9207801
  22. [23]Douglas NC, Papaioannou VE (2013) The T-box transcription factors TBX2 and TBX3 in mammary gland development and breast cancer. J Mammary Gland Biol Neoplasia 18: 143– 147 10.1007/s10911-013-9282-8 [doi] 23624936 PMC3692603 · DOI
  23. [24]Farin HF, Bussen M, Schmidt MK, Singh MK, Schuster-Gossler K, et al (2007) Transcriptional repression by the T-box proteins Tbx18 and Tbx15 depends on Groucho corepressors. J Biol Chem 282: 25748– 25759 M703724200 [pii]; 10.1074/jbc.M703724200 [doi] 17584735 · DOI
  24. [25]Rowley M, Grothey E, Couch FJ (2004) The role of Tbx2 and Tbx3 in mammary development and tumorigenesis. J Mammary Gland Biol Neoplasia 9: 109– 118. 15300007
  25. [26]Greulich F, Rudat C, Kispert A (2011) Mechanisms of T-box gene function in the developing heart. Cardiovasc Res 91: 212– 222 cvr112 [pii]; 10.1093/cvr/cvr112 [doi] 21498422 · DOI
  26. [27]King M, Arnold JS, Shanske A, Morrow BE (2006) T-genes and limb bud development. Am J Med Genet A 140: 1407– 1413 10.1002/ajmg.a.31250 [doi] 16688725 · DOI
  27. [28]Jarde T, Dale T (2011) Wnt signalling in murine postnatal mammary gland development. Acta Physiol (Oxf) 204: 118– 127. 21518264
  28. [29]Zeng YA, Nusse R (2010) Wnt proteins are self-renewal factors for mammary stem cells and promote their long-term expansion in culture. Cell Stem Cell 6: 568– 577 S1934-5909(10)00155-4 [pii]; 10.1016/j.stem.2010.03.020 [doi] 20569694 PMC2917779 · DOI
  29. [30]Brisken C, Heineman A, Chavarria T, Elenbaas B, Tan J, et al (2000) Essential function of Wnt-4 in mammary gland development downstream of progesterone signaling. Genes Dev 14: 650– 654. 10733525 PMC316462
  30. [31]Proia DA, Kuperwasser C (2006) Reconstruction of human mammary tissues in a mouse model. Nat Protoc 1: 206– 214. 17406234
  31. [32]Kuperwasser C, Chavarria T, Wu M, Magrane G, Gray JW, et al (2004) Reconstruction of functionally normal and malignant human breast tissues in mice. Proc Natl Acad Sci U S A 101: 4966– 4971. 15051869 PMC387357
  32. [33]Dontu G, Abdallah WM, Foley JM, Jackson KW, Clarke MF, et al (2003) In vitro propagation and transcriptional profiling of human mammary stem/progenitor cells. Genes Dev 17: 1253– 1270. 12756227 PMC196056
  33. [34]Nardulli AM, Katzenellenbogen BS (1988) Progesterone receptor regulation in T47D human breast cancer cells: analysis by density labeling of progesterone receptor synthesis and degradation and their modulation by progestin. Endocrinology 122: 1532– 1540 10.1210/endo-122-4-1532 [doi] 3345726 · DOI
  34. [35]Diaz J, Aranda E, Henriquez S, Quezada M, Espinoza E, et al (2012) Progesterone promotes focal adhesion formation and migration in breast cancer cells through induction of protease-activated receptor-1. J Endocrinol 214: 165– 175 JOE-11-0310 [pii]; 10.1530/JOE-11-0310 [doi] 22593082 · DOI
  35. [36]Schatz RW, Soto AM, Sonnenschein C (1985) Effects of interaction between estradiol-17 beta and progesterone on the proliferation of cloned breast tumor cells (MCF-7 and T47D). J Cell Physiol 124: 386– 390 10.1002/jcp.1041240305 [doi] 4044660 · DOI
  36. [37]Proia TA, Keller PJ, Gupta PB, Klebba I, Jones AD, et al (2011) Genetic predisposition directs breast cancer phenotype by dictating progenitor cell fate. Cell Stem Cell 8: 149– 163. 21295272 PMC3050563
  37. [38]Iyer V, Klebba I, McCready J, Arendt LM, Betancur-Boissel M, et al (2012) Estrogen promotes ER-negative tumor growth and angiogenesis through mobilization of bone marrow-derived monocytes. Cancer Res 72: 2705– 2713. 22467173
  38. [39]Keller PJ, Lin AF, Arendt LM, Klebba I, Jones AD, et al (2010) Mapping the cellular and molecular heterogeneity of normal and malignant breast tissues and cultured cell lines. Breast Cancer Res 12: R87. 20964822 PMC3096980
  39. [41]Soule HD, Maloney TM, Wolman SR, Peterson WD Jr, Brenz R, et al (1990) Isolation and characterization of a spontaneously immortalized human breast epithelial cell line, MCF-10. Cancer Res 50: 6075– 6086. 1975513
  40. [42]Stingl J, Eaves CJ, Zandieh I, Emerman JT (2001) Characterization of bipotent mammary epithelial progenitor cells in normal adult human breast tissue. Breast Cancer Res Treat 67: 93– 109. 11519870
  41. [43]Brennan KR, Brown AM (2004) Wnt proteins in mammary development and cancer. J Mammary Gland Biol Neoplasia 9: 119– 131 10.1023/B:JOMG.0000037157.94207.33 [doi];490061 [pii] 15300008 · DOI
  42. [45]Joshi PA, Jackson HW, Beristain AG, Di Grappa MA, Mote PA, et al (2010) Progesterone induces adult mammary stem cell expansion. Nature 465: 803– 807. 20445538
  43. [46]Hilton HN, Graham JD, Kantimm S, Santucci N, Cloosterman D, et al (2012) Progesterone and estrogen receptors segregate into different cell subpopulations in the normal human breast. Mol Cell Endocrinol 361: 191– 201 S0303-7207(12)00263-8 [pii]; 10.1016/j.mce.2012.04.010 [doi] 22580007 · DOI
  44. [47]Ablett MP, Singh JK, Clarke RB (2012) Stem cells in breast tumours: are they ready for the clinic? Eur J Cancer 48: 2104– 2116 S0959-8049(12)00303-6 [pii]; 10.1016/j.ejca.2012.03.019 [doi] 22542086 · DOI
  45. [48]Hilton HN, Santucci N, Silvestri A, Kantimm S, Huschtscha LI, et al.. (2014) Progesterone stimulates progenitor cells in normal human breast and breast cancer cells. Breast Cancer Res Treat. 10.1007/s10549-013-2817-2 [doi]. 24395108 · DOI
  46. [49]Couse JF, Curtis SW, Washburn TF, Lindzey J, Golding TS, et al (1995) Analysis of transcription and estrogen insensitivity in the female mouse after targeted disruption of the estrogen receptor gene. Mol Endocrinol 9: 1441– 1454. 8584021
  47. [50]Katzenellenbogen BS, Kendra KL, Norman MJ, Berthois Y (1987) Proliferation, hormonal responsiveness, and estrogen receptor content of MCF-7 human breast cancer cells grown in the short-term and long-term absence of estrogens. Cancer Res 47: 4355– 4360. 3607768
  48. [51]Huseby RA, Maloney TM, McGrath CM (1984) Evidence for a direct growth-stimulating effect of estradiol on human MCF-7 cells in vivo. Cancer Res 44: 2654– 2659. 6722798
  49. [52]Fillmore CM, Gupta PB, Rudnick JA, Caballero S, Keller PJ, et al (2010) Estrogen expands breast cancer stem-like cells through paracrine FGF/Tbx3 signaling. Proc Natl Acad Sci U S A 107: 21737– 21742. 21098263 PMC3003123
  50. [53]Clarke RB, Howell A, Potten CS, Anderson E (1997) Dissociation between steroid receptor expression and cell proliferation in the human breast. Cancer Res 57: 4987– 4991. 9371488
  51. [54]Russo J, Ao X, Grill C, Russo IH (1999) Pattern of distribution of cells positive for estrogen receptor alpha and progesterone receptor in relation to proliferating cells in the mammary gland. Breast Cancer Res Treat 53: 217– 227. 10369068
  52. [55]Mishell DR, Jr., Nakamura RM, Crosignani PG, Stone S, Kharma K, et al.. (1971) Serum gonadotropin and steroid patterns during the normal menstrual cycle. Am J Obstet Gynecol 111: 60–65. 0002-9378(71)90927-6 [pii]. 5096358
  53. [56]Uehara J, Nazario AC, Rodrigues de LG, Simoes MJ, Juliano Y, et al.. (1998) Effects of tamoxifen on the breast in the luteal phase of the menstrual cycle. Int J Gynaecol Obstet 62: 77–82. S0020729298000502 [pii]. 9722130
  54. [57]Liu J, Esmailpour T, Shang X, Gulsen G, Liu A, et al (2011) TBX3 over-expression causes mammary gland hyperplasia and increases mammary stem-like cells in an inducible transgenic mouse model. BMC Dev Biol 11: 65 1471-213X-11-65 [pii]; 10.1186/1471-213X-11-65 [doi] 22039763 PMC3217876 · DOI
  55. [58]Lim E, Wu D, Pal B, Bouras T, Asselin-Labat ML, et al (2010) Transcriptome analyses of mouse and human mammary cell subpopulations reveal multiple conserved genes and pathways. Breast Cancer Res 12: R21 bcr2560 [pii]; 10.1186/bcr2560 [doi] 20346151 PMC2879567 · DOI · Paper page
  56. [59]The Cancer Genome Atlas Network (2012) Comprehensive molcular portraits of human breast tumors. Nature 490: 61– 70. 23000897 PMC3465532

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Authors
Arendt, L. M., St. Laurent, J., Wronski, A., Caballero, S., Lyle, S. R., Naber, S. P., & Kuperwasser, C.
Affiliations
  • Tufts University (US)
  • University of Massachusetts Chan Medical School (US)
  • Tufts Medical Center (US)
Corresponding author
Kuperwasser, Charlotte
ORCID
Venue
PLoS ONE
Publisher
Public Library of Science (PLoS)
Published
2014-10-28
Volume
9
Issue
10
Pages
e111442
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0111442
PMC
PMC4211891
OpenAlex
W2103700457
Cited by
27
References
5
Citations by year
2025: 1 · 2021: 8 · 2020: 1 · 2019: 4 · 2018: 3 · 2017: 2 · 2016: 2 · 2015: 6
FWCI
1.37
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • The Journal of the American Medical Association (JAMA) Network (American Medical Association) (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Figshare (submittedVersion, cc-by)
Funders
  • Breast Cancer Research Foundation
  • National Institutes of Health (CA170851)
  • National Cancer Institute (R01CA125554, CA125554, CA170851, R01CA170851)
  • National Center for Research Resources (K01RR021858)
  • Eunice Kennedy Shriver National Institute of Child Health and Human Development (R01HD073035, HD073035)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Wnt/β-catenin signaling in development and cancer
  • Health Sciences › Medicine › Oncology › Cancer Cells and Metastasis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Estrogen and related hormone effects
Keywords
Wnt signaling, TBX3, Wnt1, LRP6, mammary epithelial cells, estrogen, progesterone, paracrine signaling, autocrine signaling, breast development, breast reduction
MeSH terms
Animals, Breast, Collagen, Epithelial Cells, Estrogens, Female, Humans, Ligands, Phenotype, Progesterone, Stem Cells, Signal Transduction, Lentivirus, Mice, SCID, Mice, Inbred NOD, T-Box Domain Proteins, RNA, Small Interfering, Cell Line, Tumor, Cell Proliferation, Wnt Proteins, Mice, Low Density Lipoprotein Receptor-Related Protein-6, Primary Cell Culture

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