Skip to main content
SpringerLink
Log in

Characterization of a novel silencer element in the human aromatase gene PII promoter

  • Published:
Breast Cancer Research and Treatment Aims and scope Submit manuscript

Abstract

Approximately two thirds of breast cancer patients have estrogen-dependent carcinomas. The biosynthesis of estrogens is catalyzed by the microsomal enzyme aromatase. Mechanisms controlling human aromatase gene expression are complicated by the existence of multiple tissue specific promoters. The most proximally located PII promoter is mainly active in ovarian granulosa cells. PII can be switched on in human breast cancer cells. Since there are strong silencer elements located within the 3′ portion of the PII promoter, we propose that the function of these silencer elements could be reversed by breast cancer cell specific signals/factors, resulting in aberrant expression of aromatase. We have identified and characterized a novel silencer element, S2, which is upstream of S1, a silencer element recently identified by another group. S2, a 54-bp fragment 100% conserved between humans and rodents, functions in both orientation- and promoter-independent manners. The core region of S2 contains two consensus binding sites for members of the GATA transcription factors. GATA-4 was found to be expressed in three out of four human breast cancer cell lines examined by RT-PCR, and transfection with GATA-4 partially reversed the repressive function of S2. However, we were unable to demonstrate that DNA-protein complexes formed between nuclear extracts of human breast and ovarian cancer cells and S2 contain GATA-4 using a supershifting approach. We suggest that the expression of GATA-4, and more importantly, other yet to be identified GATA or GATA-related factor(s), are implicated in provoking aberrant expression of aromatase, and therefore, the biosynthesis of estrogens, in human breast cancer cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Nass SJ, Hahm HA, Davidson NE: Breast cancer biology blossoms in the clinic. Nat Med 4: 761–762, 1998

    PubMed  Google Scholar 

  2. Simpson ER, Mahendroo MS, Means GD, Kilgore MW, Hinshelwood MM, Graham-Lorence SG, Amarneh B, Ito Y, Fisher GR, Michael MD, Mendelson CR, Bulun SE: Aromatase cytochrome P450, the enzyme responsible for estrogen biosynthesis. Endocrine Rev 15: 342–355, 1994

    Google Scholar 

  3. Corbin CJ, Graham-Lorence S, McPhaul M, Mason JI, Mendelson CR, Simpson ER: Isolation of a full length cDNA insert encoding human aromatase system cytochrome P-450 and its expression in non-steroidogenic cells. Proc Natl Acad Sci USA, 85: 8948–8952, 1988

    PubMed  Google Scholar 

  4. Harada N: Cloning of a comparative cDNA encoding human aromatase: immunochemical identification and sequence analysis. Biochem Biophys Res Commun 156: 725–732, 1988

    PubMed  Google Scholar 

  5. Means, GD, Kilgore MW, Mahendroo MS, Mendelson CR, Simpson ER: Tissue-specific promoters regulate aromatase cytochrome P450 gene expression in human ovary and fetal tissues. Mol Endocrinol 5: 2005–2013, 1991

    PubMed  Google Scholar 

  6. Kilgore MW, Means GD, Mendelson CR, Simpson ER: Alternative promotion of aromatase P-450 expression in the human fetal tissue. Mol Cell Endocrinol 83: R9–R16, 1992

    PubMed  Google Scholar 

  7. Zhao Y, Nichols JE, Bulum SE, Mendelson CR, Simpson ER: Aromatase P450 gene expression in human adipose tissue. Role of a Jak/STAT in regulation of the adipose-specific promoter. J Biol Chem 270: 16449–16457, 1995

    PubMed  Google Scholar 

  8. Esteban JM, Warsi Z, Haniu M, Hall P, Shively JE, Chen S: Detection of intratumoral aromatase in breast carcinomas. An immunohistochemical study with clinicopathologic correlation. Am J Pathol 140: 337–343, 1992

    PubMed  Google Scholar 

  9. Koos RD, Banks PK, Inkster SE, Yue W, Brodie AM: Detection of aromatase and keratinocyte growth factor expression in breast tumors using reverse transcription-polymerase chain reaction. J Steroid Biochem Mol Biol 45: 217–225, 1993

    PubMed  Google Scholar 

  10. Zhou C, Zhou D, Esteban J, Murai J, Sitteri PI, Wilczynski A, Chen S: Aromatase gene expression and its exon I usage in human breast tumors. Detection of aromatase messenger RNA by reverse transcription-polymerase chain reaction (RT-PCR). J Steroid Biochem Mol Biol 59: 163–171, 1996

    PubMed  Google Scholar 

  11. Zhou D, Clarke P, Wang J, Chen S: Identification of a promoter that controls aromatase expression in human breast cancer and adipose stromal cells. J Biol Chem 271, 15194–15202, 1996

    PubMed  Google Scholar 

  12. Yue W, Wang JP, Hamilton CJ, Demers LM, Santen RJ: In situ aromatization enhances breast tumor estradiol levels and cellular proliferation. Cancer Res 58: 927–932, 1998

    PubMed  Google Scholar 

  13. Wang J, Chen S: Identification of a promoter and a silencer at the 3′-end of the first intron of the human aromatase gene. Mol Endocrinol 6: 1479–1488, 1992

    PubMed  Google Scholar 

  14. Zhou D, Chen S: Characterization of a silencer element in the human aromatase gene. Archives Biochem Biophys 315: 213–220, 1998

    Google Scholar 

  15. Yang C, Zhou D, Chen S:Modulation of aromatase expression in the breast tissue by ERRa-1 orphan receptor. Cancer Res 58: 5695–5700, 1998

    PubMed  Google Scholar 

  16. Jin T, Branch DR, Zhang X, Qi S, Youngson B, Goss PE: Examination of POU homeobox gene expression in human breast cancer cells. Int J Cancer 81: 104–112, 1999

    PubMed  Google Scholar 

  17. Jin T, Drucker DJ: Activation of proglucagon gene transcription through a novel G1 promoter element by the caudalrelated homeodomain protein cdx-2/3. Mol Cell Biol 16: 19–28, 1996

    PubMed  Google Scholar 

  18. Mills GB, May C, Hill M, Campbell S, Shaw P, Marks A: Ascitic fluid from human ovarian cancer patients contains growth factors necessary for intraperitoneal growth of human ovarian adenocarcinoma cells. J Clin Invest 86: 851–585, 1990

    PubMed  Google Scholar 

  19. Arceci RJ, King AAJ, Simon MC, Orkin SH, Wilson DB: Mouse GATA-4: a retinoic acid-inducible GATA-binding transcription factor expressed in endodermally derived tissues and heart. Mol Cell Biol 13: 2235–2246, 1993

    PubMed  Google Scholar 

  20. Jin T, Drucker DJ: The proglucagon gene upstream enhancer (GUE) contains positive and negative domains important for tissue-specific proglucagon gene transcription. Mol Endocrinol 9: 1306–1320, 1995

    PubMed  Google Scholar 

  21. Schreiber E, Matthias P, Muller MM, Schaffner W: Rapid detection of octamer binding proteins with ‘mini-extracts’ prepared from a small number of cells. Nucleic Acids Res 17: 6419, 1989

    PubMed  Google Scholar 

  22. Hickey GJ, Krasnow JF, Beattie WG, Richards JS: Aromatase cytochrome P450 in rat ovarian granulosa cells before and after luteinization: adenosine 3′,5′-monophosphate-dependent and independent regulation. Cloning and sequencing of rat aromatase cDNA and 5′ genomic DNA. Mol Endocrinol 4: 3–12, 1990

    PubMed  Google Scholar 

  23. Fitzpatrick SL, Richards JS: Cis-acting elements of the rat aromatase promoter required for cyclic adenosine 3′,5′-monophosphate induction in ovarian granulosa cells and constitutive expression in R2C leydig cells. Mol Endocrinol 7: 341–354, 1993

    PubMed  Google Scholar 

  24. Gonzalez-Robayna IJ, Alliston TN, Buse P, Firestone GL, Richards JS: Functional and sub-cellular changes in the Akinase-signalingpathway: relation to aromatase and Sgk expression during the transition of granulosa cells to luteal cells. Mol Endocrinol 13: 1318–1337, 1999

    PubMed  Google Scholar 

  25. Tsai SF, Martin DIK, Zon LI, D'Andrea AD, Wong GG, Orkin SH: Cloning of cDNA for the major DNA binding protein of the erythroid lineage through expression in mammalian cells. Nature 339: 446–451, 1989

    PubMed  Google Scholar 

  26. Weiss MJ, Orkin SH: GATA transcription factors: key regulators of hematopoiesis. Exp Hematol 23: 99–107, 1995

    PubMed  Google Scholar 

  27. Yamagata T, Nishida J, Sakai R, Tanaka T, Honda H, Hirano N, Mano H, Yazaki Y, Hirai H: Of the GATA-binding proteins, only GATA-4 selectively regulates the human interleukin-5 gene promoter in interleukin-5-producing cells which express multiple GATA-binding proteins. Mol Cell Biol 15: 3830–3839, 1995

    PubMed  Google Scholar 

  28. Jiang Y, Tarzami S, Burch JB, Evans T: Common role for each of the cGATA-4/5/6 genes in the regulation of cardiac morphogenesis. Dev Genet 22: 263–277, 1998

    PubMed  Google Scholar 

  29. Gao X, Sedgwick T, Shi YB, Evans T: Distinct functions are implicated for the GATA-4,-5, and-6 transcription factors in the regulation of intestine epithelial cell differentiation. Mol Cell Biol 18: 2901–2911, 1998

    PubMed  Google Scholar 

  30. Harada N: A unique aromatase (P-450AROM) mRNA formed by alternative use of tissue-specific exons 1 in human skin fibroblasts. Biochem Biophys Res Commun 189: 1001–1007, 1992

    PubMed  Google Scholar 

Download references

Authors
  1. Tianru Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiaoyun Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Huiqin Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Paul E. Goss
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jin, T., Zhang, X., Li, H. et al. Characterization of a novel silencer element in the human aromatase gene PII promoter. Breast Cancer Res Treat 62, 151–159 (2000). https://doi.org/10.1023/A:1006481228794

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006481228794

Search

Navigation