Tumor Biology

, Volume 31, Issue 1, pp 16–22

Estrogen prevents sustained COLO-205 human colon cancer cell growth by inducing apoptosis, decreasing c-myb protein, and decreasing transcription of the anti-apoptotic protein bcl-2

  • Heather R. Wilkins
  • Kristin Doucet
  • Victoria Duke
  • Amber Morra
  • Nicole Johnson
Research Article

Abstract

The proto-oncogene c-myb is overexpressed in human colon cancer cells. c-myb is known to be affected by estrogen in some breast cancers and leukemias. However, the mechanism of c-myb regulation via estrogen in colon cancer requires further investigation. Human COLO-205 colon cancer cells were cultured and treated with beta-estradiol for 24 h. Apoptosis was quantified using acridine orange/propidium iodide labeling and confirmed with DNA fragmentation gel electrophoresis. Expression of c-myb protein was assessed via SDS-PAGE and immunoblotting and RT-PCR was used to quantify bcl-2 RNA. Protein and RNA expression levels were also assayed after c-myb siRNA treatment for 24 h. We demonstrate an increase in apoptosis after 24 h of β-estradiol treatment of human COLO-205 colon cancer cells. Estrogen treatment also decreases c-myb protein levels as well as expression of its transcriptional target bcl-2. Suppression of c-myb protein also results in increased apoptosis and decreases bcl-2 expression. These results indicate that estrogen has a protective effect from sustained colon cancer cell growth at least partly through suppression of c-myb and bcl-2.

Keywords

c-myb Estrogen COLO-205 bcl-2 Apoptosis 

References

  1. 1.
    Jagani Z, Khosravi-Far R. Cancer stem cells and impaired apoptosis. Adv Exp Med Biol. 2008;615:331–44.CrossRefPubMedGoogle Scholar
  2. 2.
    Aggarwal BB, Ichikawa H. Molecular targets and anticancer potential of indole-3-carbinol and its derivatives. Cell Cycle. 2005;4:1201–15.PubMedGoogle Scholar
  3. 3.
    Lechner D, Kallay E, Cross HS. Phytoestrogens and colorectal cancer prevention. Vitam Horm. 2005;70:169–98.CrossRefPubMedGoogle Scholar
  4. 4.
    Liu X, Lazenby AJ, Siegal GP. Signal transduction cross-talk during colorectal tumorigenesis. Adv Anat Pathol. 2006;13:270–4.CrossRefPubMedGoogle Scholar
  5. 5.
    Jakowlew SB. Transforming growth factor-beta in cancer and metastasis. Cancer Metastasis Rev. 2006;25:435–57.CrossRefPubMedGoogle Scholar
  6. 6.
    Ramocki NM, Wilkins HR, Magness ST, Simmons JG, Scull BP, Lee GH, et al. Insulin receptor substrate-1 deficiency promotes apoptosis in the putative intestinal crypt stem cell region, limits apcmin/+ tumors, and regulates sox9. Endocrinology. 2008;149:261–7.CrossRefPubMedGoogle Scholar
  7. 7.
    Torelli G, Venturelli D, Colo A, Zanni C, Selleri L, Moretti L, et al. Expression of c-myb protooncogene and other cell cycle-related genes in normal and neoplastic human colonic mucosa. Cancer Res. 1987;47:5266–9.PubMedGoogle Scholar
  8. 8.
    Greco C, Alvino S, Buglioni S, Assisi D, Lapenta R, Grassi A, et al. Activation of c-myc and c-myb proto-oncogenes is associated with decreased apoptosis in tumor colon progression. Anticancer Res. 2001;21:3185–92.PubMedGoogle Scholar
  9. 9.
    Funato T, Satou J, Kozawa K, Fujimaki S, Miura T, Kaku M. Use of c-myb antisense oligonucleotides to increase the sensitivity of human colon cancer cells to cisplatin. Oncol Rep. 2001;8:807–10.PubMedGoogle Scholar
  10. 10.
    Frampton J, Ramqvist T, Graf T. V-myb of e26 leukemia virus up-regulates bcl-2 and suppresses apoptosis in myeloid cells. Genes Dev. 1996;10:2720–31.CrossRefPubMedGoogle Scholar
  11. 11.
    Taylor D, Badiani P, Weston K. A dominant interfering myb mutant causes apoptosis in t cells. Genes Dev. 1996;10:2732–44.CrossRefPubMedGoogle Scholar
  12. 12.
    Gupta S, Kass GE, Szegezdi E, Joseph B The mitochondrial death pathway: A promising therapeutic target in diseases. J Cell Mol Med. 2009 (in press)Google Scholar
  13. 13.
    Kim R, Emi M, Tanabe K. Role of mitochondria as the gardens of cell death. Cancer Chemother Pharmacol. 2006;57:545–53.CrossRefPubMedGoogle Scholar
  14. 14.
    Gonda TJ, Leo P, Ramsay RG. Estrogen and myb in breast cancer: potential for new therapies. Expert Opin Biol Ther. 2008;8:713–17.CrossRefPubMedGoogle Scholar
  15. 15.
    Drabsch Y, Hugo H, Zhang R, Dowhan DH, Miao YR, Gewirtz AM, et al. Mechanism of and requirement for estrogen-regulated myb expression in estrogen receptor-positive breast cancer cells. Proc Natl Acad Sci U S A. 2007;104:13762–7.CrossRefPubMedGoogle Scholar
  16. 16.
    Gudas JM, Klein RC, Oka M, Cowan KH. Posttranscriptional regulation of the c-myb proto-oncogene in estrogen receptor-positive breast cancer cells. Clin Cancer Res. 1995;1:235–43.PubMedGoogle Scholar
  17. 17.
    Humphries KH, Gill S. Risks and benefits of hormone replacement therapy: the evidence speaks. Cmaj. 2003;168:1001–10.PubMedGoogle Scholar
  18. 18.
    Lemay A. The relevance of the women's health initiative results on combined hormone replacement therapy in clinical practice. J Obstet Gynaecol Can. 2002;24:711–15.PubMedGoogle Scholar
  19. 19.
    Grodstein F, Newcomb PA, Stampfer MJ. Postmenopausal hormone therapy and the risk of colorectal cancer: a review and meta-analysis. Am J Med. 1999;106:574–82.CrossRefPubMedGoogle Scholar
  20. 20.
    Hsu HH, Cheng SF, Wu CC, Chu CH, Weng YJ, Lin CS, et al. Apoptotic effects of over-expressed estrogen receptor-beta on lovo colon cancer cell is mediated by p53 signalings in a ligand-dependent manner. Chin J Physiol. 2006;49:110–16.PubMedGoogle Scholar
  21. 21.
    Qiu Y, Langman MJ, Eggo MC. Targets of 17beta-oestradiol-induced apoptosis in colon cancer cells: a mechanism for the protective effects of hormone replacement therapy? J Endocrinol. 2004;181:327–37.CrossRefPubMedGoogle Scholar
  22. 22.
    Qiu Y, Waters CE, Lewis AE, Langman MJ, Eggo MC. Oestrogen-induced apoptosis in colonocytes expressing oestrogen receptor beta. J Endocrinol. 2002;174:369–77.CrossRefPubMedGoogle Scholar
  23. 23.
    Motylewska E, Lawnicka H, Melen-Mucha G. Oestradiol and tamoxifen inhibit murine colon 38 cancer growth and increase the cytotoxic effect of fluorouracil. Endokrynol Pol. 2007;58:426–34.PubMedGoogle Scholar
  24. 24.
    Thompson MA, Rosenthal MA, Ellis SL, Friend AJ, Zorbas MI, Whitehead RH, et al. C-myb down-regulation is associated with human colon cell differentiation, apoptosis, and decreased bcl-2 expression. Cancer Res. 1998;58:5168–75.PubMedGoogle Scholar
  25. 25.
    Wolff L, Schmidt M, Koller R, Haviernik P, Watson R, Bies J, et al. Three genes with different functions in transformation are regulated by c-myb in myeloid cells. Blood Cells Mol Dis. 2001;27:483–8.CrossRefPubMedGoogle Scholar
  26. 26.
    Blick M, Westin E, Gutterman J, Wong-Staal F, Gallo R, McCredie K, et al. Oncogene expression in human leukemia. Blood. 1984;64:1234–9.PubMedGoogle Scholar
  27. 27.
    Gonda TJ. The c-myb oncoprotein. Int J Biochem Cell Biol. 1998;30:547–51.CrossRefPubMedGoogle Scholar
  28. 28.
    Liang W. Age, sex and the risk of grade-specific second primary colorectal cancer: evidence for the protective effect of female hormone. Eur J Cancer. 2007;43:1856–61.CrossRefPubMedGoogle Scholar
  29. 29.
    Yu Z, Li W, Liu F. Inhibition of proliferation and induction of apoptosis by genistein in colon cancer ht-29 cells. Cancer Lett. 2004;215:159–66.CrossRefPubMedGoogle Scholar
  30. 30.
    Couse JF, Lindzey J, Grandien K, Gustafsson JA, Korach KS. Tissue distribution and quantitative analysis of estrogen receptor-alpha (eralpha) and estrogen receptor-beta (erbeta) messenger ribonucleic acid in the wild-type and eralpha-knockout mouse. Endocrinology. 1997;138:4613–21.CrossRefPubMedGoogle Scholar
  31. 31.
    Kennelly R, Kavanagh DO, Hogan AM, Winter DC. Oestrogen and the colon: potential mechanisms for cancer prevention. Lancet Oncol. 2008;9:385–91.CrossRefPubMedGoogle Scholar
  32. 32.
    Kuiper GG, Shughrue PJ, Merchenthaler I, Gustafsson JA. The estrogen receptor beta subtype: a novel mediator of estrogen action in neuroendocrine systems. Front Neuroendocrinol. 1998;19:253–86.CrossRefPubMedGoogle Scholar
  33. 33.
    Ellem SJ, Risbridger GP. The dual, opposing roles of estrogen in the prostate. Ann N Y Acad Sci. 2009;1155:174–86.CrossRefPubMedGoogle Scholar
  34. 34.
    Thornton MJ. The biological actions of estrogens on skin. Exp Dermatol. 2002;11:487–502.CrossRefPubMedGoogle Scholar
  35. 35.
    Dechering K, Boersma C, Mosselman S. Estrogen receptors alpha and beta: two receptors of a kind? Curr Med Chem. 2000;7:561–76.PubMedGoogle Scholar
  36. 36.
    Kuiper GG, Enmark E, Pelto-Huikko M, Nilsson S, Gustafsson JA. Cloning of a novel receptor expressed in rat prostate and ovary. Proc Natl Acad Sci U S A. 1996;93:5925–30.CrossRefPubMedGoogle Scholar
  37. 37.
    Tremblay GB, Tremblay A, Copeland NG, Gilbert DJ, Jenkins NA, Labrie F, et al. Cloning, chromosomal localization, and functional analysis of the murine estrogen receptor beta. Mol Endocrinol. 1997;11:353–65.CrossRefPubMedGoogle Scholar
  38. 38.
    Mosselman S, Polman J, Dijkema R. Er beta: Identification and characterization of a novel human estrogen receptor. FEBS Lett. 1996;392:49–53.CrossRefPubMedGoogle Scholar
  39. 39.
    Girdler F, Brotherick I. The oestrogen receptors (er alpha and er beta) and their role in breast cancer: a review. Breast. 2000;9:194–200.CrossRefPubMedGoogle Scholar
  40. 40.
    Strobl JS, Wonderlin WF, Flynn DC. Mitogenic signal transduction in human breast cancer cells. Gen Pharmacol. 1995;26:1643–9.PubMedGoogle Scholar
  41. 41.
    Verdier-Sevrain S, Yaar M, Cantatore J, Traish A, Gilchrest BA. Estradiol induces proliferation of keratinocytes via a receptor-mediated mechanism. Faseb J. 2004;18:1252–4.PubMedGoogle Scholar
  42. 42.
    Qi X, Borowicz S, Pramanik R, Schultz RM, Han J, Chen G. Estrogen receptor inhibits c-jun-dependent stress-induced cell death by binding and modifying c-jun activity in human breast cancer cells. J Biol Chem. 2004;279:6769–77.CrossRefPubMedGoogle Scholar
  43. 43.
    Ramsay RG, Gonda TJ. Myb function in normal and cancer cells. Nat Rev Cancer. 2008;8:523–34.CrossRefPubMedGoogle Scholar
  44. 44.
    Thompson MA, Flegg R, Westin EH, Ramsay RG. Microsatellite deletions in the c-myb transcriptional attenuator region associated with over-expression in colon tumour cell lines. Oncogene. 1997;14:1715–23.CrossRefPubMedGoogle Scholar
  45. 45.
    Jakacka M, Ito M, Weiss J, Chien PY, Gehm BD, Jameson JL. Estrogen receptor binding to DNA is not required for its activity through the nonclassical ap1 pathway. J Biol Chem. 2001;276:13615–21.PubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2009

Authors and Affiliations

  • Heather R. Wilkins
    • 1
  • Kristin Doucet
    • 1
  • Victoria Duke
    • 1
  • Amber Morra
    • 1
  • Nicole Johnson
    • 1
  1. 1.Department of Natural SciencesAssumption CollegeWorcesterUSA

Personalised recommendations