Skip to main content
SpringerLink
Log in

Derepression of HMGA2 Gene Expression in Retinoblastoma Is Associated with Cell Proliferation

  • Articles
  • Published:
Molecular Medicine Aims and scope Submit manuscript

Abstract

To assess whether retinoblastoma formation is associated with the expression of high mobility group (HMG) A2 protein, a transcription factor that is highly expressed during embryogenesis and completely repressed in normal adult tissues, we performed Northern and Western blots and RT-PCR analyses, and immunohistochemistry to test for HMGA2 expression. We used established retinoblastoma cell lines in tumors grown in nude mice and clinical retinoblastoma specimens, and contrasted these tumors with normal embryonic and adult retina. Adenoviral-mediated antisense experiments were conducted on the retinoblastoma cell lines to suppress HMGA2 expression and determine if cell proliferation is HMGA2-dependent. We also transfected a retinoblastoma cell line to identify cis-regulatory elements and transcription initiation sites on the HMGA2 gene promoter. HMGA2 gene expression was silenced in terminally differentiated retina of 6-wk-old mice, but it was detected in retina of a 13.5-d postcoitum embryo. Reactivation of HMGA2 gene expression was observed in the retinoblastoma cell lines Y79, WERI-Rb1, and TOTL-1, in tumors derived from some of these cells propagated in nude mice, and in a high frequency of retinoblastomas excised from human patients. This suggests that expression of HMGA2 gene in retinoblastoma cells involves a derepression process. By using an antisense approach to block HMGA2 expression, we observed a decrease in the number of proliferating retinoblastoma cells. As a 1st step toward understanding HMGA2 gene reactivation in retinoblastoma, we mapped the 2 transcription initiation sites and associated positive regulatory elements within the WERI-Rb1 cells. Our discovery of derepression of HMGA2 gene expression in retinoblastoma provides the 1st evidence that this protein might contribute to neoplastic transformation of retina 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

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

References

  1. Albert DA, Jakobiec FA (eds). (1994) Principles and practice of ophthalmology. Sauders, Philadelphia.

    Google Scholar 

  2. Friend SH et al. (1986) A human DNA segment with properties of the gene that predisposes to retinoblastoma and osteosarcoma. Nature 323:643–6.

    Article  CAS  Google Scholar 

  3. Lee WH et al. (1987) Human retinoblastoma susceptibility gene: cloning, identification, and sequence. Science 235:1394–9.

    Article  CAS  Google Scholar 

  4. Fung YK et al. (1987) Structural evidence for the authenticity of the human retinoblastoma gene. Science 236:1657–61.

    Article  CAS  Google Scholar 

  5. Goodwin G. (1998) Molecules in focus: the high mobility group protein, HMGI-C. Int J Biochem. Cell Biol. 30:761–6.

    Article  CAS  Google Scholar 

  6. Zhou X, Chada K. (1998) HMGI family proteins: architectural transcription factors in mammalian development and cancer. Keio J. Med. 47:73–7.

    Article  CAS  Google Scholar 

  7. Liu F, Chau KY, Arlotta P, Ono SJ. (2001) The HMG I proteins: dynamic roles in gene activation, development, and tumorigenesis. Immunol. Res. 24:13–29.

    Article  Google Scholar 

  8. Reeves R. (2001) Molecular biology of HMGA proteins: hubs of nuclear function. Gene 277:63–81.

    Article  CAS  Google Scholar 

  9. Fedele M et al. (2001) Role of the high mobility group A proteins in human lipomas. Carcinogenesis 22:1583–91.

    Article  CAS  Google Scholar 

  10. Wolffe A. (1994) Architectural transcription factors. Science 264:1100–1.

    Article  CAS  Google Scholar 

  11. Mantovani F et al. (1998) NF-κB mediated transcriptional activation is enhanced by the architectural factor HMGI-C. Nucleic Acids Res. 26:1433–9.

    Article  CAS  Google Scholar 

  12. Noro B et al. (2003) Molecular dissection of the architectural transcription factor HMGA2. Biochemistry 42:4569–77.

    Article  CAS  Google Scholar 

  13. Zhou X et al. (1995) Mutation responsible for the mouse pygmy phenotype in the developmentally regulated factor HMGI-C. Nature 376:771–4.

    Article  CAS  Google Scholar 

  14. Zhou X et al. (1996) Genomic structure and expression of the murine HMGI-C gene. Nucleic Acids Res. 24:4071–7.

    Article  CAS  Google Scholar 

  15. Rogalla P et al. (1996) HMGI-C expression patterns in human tissues. implications for the genesis of frequent mesenchymal tumors. Am. J. Pathol. 149:775–9.

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Hirning-Folz U et al. (1998) The expression pattern of the HMGIC gene during development. Gene Chrom. Cancer 23:771–4.

    Article  Google Scholar 

  17. Chieffi P et al. (2002) HMGA1 and HMGA2 protein expression in mouse spermatogenesis. Oncogene 21:3644–50.

    Article  CAS  Google Scholar 

  18. Giancotti V et al. (1985) Changes in nuclear proteins following transformation of rat thyroid epithelial cells by a murine sarcoma retrovirus. Cancer Res. 45:6051–7.

    CAS  PubMed  Google Scholar 

  19. Giancotti V et al. (1987) Elevated levels of a specific class of nuclear phospho-proteins in cells transformed with v-ras and v-mos oncogenes and by co-transfection with c-myc and polyma middle T genes. EMBO J. 6:1981–7.

    Article  CAS  Google Scholar 

  20. Giancotti V et al. (1991) Comparison of multiple forms of the high mobility group I proteins in rodent and human cells. Identification of the human high mobility group I-C protein. Eur. J. Biochem. 198:211–6.

    Article  CAS  Google Scholar 

  21. Manfioletti G et al. (1991) cDNA cloning of the HMGI-C phosphoprotein, a nuclear protein associated with neoplastic and undifferentiated phenotypes. Nucleic Acids Res. 19:6793–7.

    Article  CAS  Google Scholar 

  22. Patel UA et al. (1994) Expression and cDNA cloning of human HMGI-C phos-phoprotein. Biochem. Biophys. Res. Commun. 201:63–70.

    Article  CAS  Google Scholar 

  23. Rogalla P et al. (1997) Expression of HMGI-C, a member of the high mobility group protein family, in a subset of breast cancers: relationship to histologic grade. Mol. Carcinog. 19:153–6.

    Article  CAS  Google Scholar 

  24. Rogalla P et al. (1998) HMGIC expression patterns in non-small lung cancer and surrounding tissue. Anticancer Res. 18:3327–30.

    CAS  PubMed  Google Scholar 

  25. Finelli P et al. (2002) The High Mobility Group A2 gene is amplified and overexpressed in human prolactinomas. Cancer Res. 62:2398–405.

    CAS  PubMed  Google Scholar 

  26. Wunderlich V, Bottger M. (1997) High-mobility-group proteins and cancer: an emerging link. J. Cancer Res. Clin. Oncol. 123:133–40.

    CAS  PubMed  Google Scholar 

  27. Wood LJ, Maher JF, Bunton TE, Resar LM. (2000) The oncogenic properties of the HMG-I gene family. Cancer Res. 60:4256–61.

    CAS  PubMed  Google Scholar 

  28. Fedele M et al. (2002) Overexpression of the HMGA2 gene in transgenic mice leads to the onset of pituitary adenomas. Oncogene 21:3190–8.

    Article  CAS  Google Scholar 

  29. Schoenmakers EFPM et al. (1995) Recurrent rearrangements in the high mobility group protein gene, HMGI-C, in benign mesenchymal tumours. Nature Genet. 10:436–44.

    Article  CAS  Google Scholar 

  30. Ashar HR et al. (1995) Disruption of the architectural factor HMGI-C: DNA-binding AT hook motifs fused in lipomas to distinct transcriptional regulatory domains. Cell. 82:57–65.

    Article  CAS  Google Scholar 

  31. Kazmierczak B et al. (1996) HMG I-C rearrangements as the molecular basis for the majority of pulmonary chondroid hamartomas: a survey of 30 tumors. Oncogene 12:515–21.

    CAS  PubMed  Google Scholar 

  32. Berner JM et al. (1997) HMGIC, the gene for an architectural transcription factor, is amplified and rearranged in a subset of human sarcomas. Oncogene 14:2935–41.

    Article  CAS  Google Scholar 

  33. Pedeutour F et al. (2000) Dysregulation of HMGIC in a uterine lipoleiomyoma with a complex rearrangement including chromosomes 7, 12, and 14. Genes Chrom. Cancer 27:209–15.

    Article  CAS  Google Scholar 

  34. Finelli P et al. (2002) The High Mobility Group A2 gene is amplified and overex-pressed in human prolactinomas. Cancer Res. 62:2398–405.

    CAS  PubMed  Google Scholar 

  35. Fedele M et al. (1998) Truncated and chimeric HMGI-C genes induce neoplastic transformation of NIH3T3 murine fibroblasts. Oncogene 17:413–8.

    Article  CAS  Google Scholar 

  36. Battista S et al. (1999) The expression of a truncated HMGI-C gene induces gigantism associated with lipomatosis. Cancer Res. 59:4793–7.

    CAS  PubMed  Google Scholar 

  37. Arlotta P et al. (2000) Transgenic mice expressing a truncated form of the high mobility group i-c protein develop adiposity and an abnormally high prevalence of lipomas. J. Biol. Chem. 275:14394–400.

    Article  CAS  Google Scholar 

  38. Chau KY et al. (2000) The architectural transcription factor high mobility group I(Y) participates in photoreceptor-specific gene expression. J Neurosci. 20:7317–24.

    Article  CAS  Google Scholar 

  39. Sasabe T et al. (1991) Cyclic nucleotides and differentiation of retinoblastoma. Folia Ophthalmol. 42:1200–6.

    Google Scholar 

  40. Vanhamme L, Szpirer C. (1988) Transforming activity of the human mammary line HBL100 DNA is associated with SV40 T antigen genetic information integrated in its genome. Carcinogenesis 9:653–5.

    Article  CAS  Google Scholar 

  41. Johnson KR et al. (1988) Complete murine cDNA sequence, genomic structure, and tissue expression of the high mobility group protein HMG-I(Y). J. Biol. Chem. 263:18338–42.

    CAS  PubMed  Google Scholar 

  42. Chiappetta G et al. (1996) High level expression of the HMGI(Y) gene during embryonic development. Oncogene 13:2439–46.

    CAS  PubMed  Google Scholar 

  43. Scala S et al. (2000) Adenovirus-mediated suppression of HMGI(Y) protein synthesis as potential therapy of human malignant neoplasias. Proc. Natl. Acad. Sci. U.S.A. 97:4256–61.

    Article  CAS  Google Scholar 

  44. Chau KY, Ono SJ. (1999) Gene transfer into retinoblastoma cells. Biotechniques 26:444–6.

    Article  CAS  Google Scholar 

  45. Chau K et al. (1999) A novel downstream positive regulatory element mediating transcription of the human high mobility group (HMG) I-C gene. FEBS Lett. 457:429–36.

    Article  CAS  Google Scholar 

  46. Berlingieri MT et al. (1995) Inhibition of HMGI-C protein synthesis suppresses retrovirally induced neoplastic transformation of rat thyroid cells. Mol. Cell Biol. 15:1545–53.

    Article  CAS  Google Scholar 

  47. White JB, Taylor RE, Pittler SJ. (2001) Reproducible high efficiency gene transfer into Y79 retinoblastoma cells using adenofection. J. Neurosci. Methods 106:1–7.

    Article  CAS  Google Scholar 

  48. Chau K-Y et al. (1995) The gene for the human architectural transcription factor HMGI-C consists of five exons each coding for a distinct functional element. Nucleic Acids Res. 23:4262–6.

    Article  CAS  Google Scholar 

  49. Prestridge DS. (1991) Signal scan: a computer program that scan DNA sequences for eukaryotic transcriptional elements. CABIOS 7:203–6.

    CAS  PubMed  Google Scholar 

  50. Ishii S et al. (1985) Characterization and sequence of the promoter region of the human epidermal growth factor receptor gene. Proc. Natl. Acad. Sci. U.S.A. 82:4920–4.

    Article  CAS  Google Scholar 

  51. Friedmann M et al. (1993) Organization, inducible-expression and chromosome localization of the human HMG-I(Y) nonhistone protein gene. Nucleic Acids Res. 21:4259–67.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We thank Drs Gilbert Jay, C Kathleen Dorey, Patricia Pearson, and other members of our laboratory, in particular: Drs Andrea Keane-Myers, Dai Miyazaki, Paola Arlotta, Takao Nakamura, and Jason Kreisberg who provided advice and help with the manuscript. We also thank Dr Robert Weinberg, Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, for the gifts of Y79, WERI-Rb1, and OSH50T cells; and the Tissue Culture Core Facility of the Lombardi Cancer Center for HBL–100 cells. This work was supported by National Institute of General Medical Science grant RO1 GM49661, the Lucille P Markey Charitable Trust, the Research to Prevent Blindness Foundation, and the “Associazione Italiana per la Ricerca sul Cancro.” GM was supported by “Fondazione Italiana per la Ricerca sul Cancro” while he was a Visiting Scientist of the Ono Lab. SJO was a JSPS Visiting Professor at Kyoto University during the preparation of this manuscript. K-YC was a Fellow of the Research to Prevent Blindness, America Fund (PD97054). SM was supported by the Massachusetts Lions Eye Research Fund and Gifts to the Retina Genetics Fund, Massachusetts Eye & Ear Infirmary.

Author information

Authors and Affiliations

  1. Department of Immunology, Institutes of Ophthalmology and Child Health, University College London, University of London, London, UK

    Kai-Yin Chau, Kam-Wa Cheung-Chau & Santa Jeremy Ono

  2. Dipartimento di Biochimica, Biofisica e Chimica delle Macromolecule, Universitá di Trieste, Trieste, Italy

    Guidalberto Manfioletti

  3. Dipartimento di Biologia e Patologia Cellulare e Molecolare, Universitá degli Studi di Napoli “Federico II,”, Napoli, Italy

    Alfredo Fusco

  4. Developmental Biology Unit, Institute of Child Health, University College London, London, UK

    Nathalie Dhomen & Jane C. Sowden

  5. Department of Ophthalmology, Osaka Habikino Hospital, Osaka, Japan

    Tetsuo Sasabe

  6. Massachusetts Eye & Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, MA, USA

    Shizuo Mukai

Authors
  1. Kai-Yin Chau
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Guidalberto Manfioletti
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kam-Wa Cheung-Chau
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Alfredo Fusco
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Nathalie Dhomen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jane C. Sowden
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Tetsuo Sasabe
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Shizuo Mukai
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Santa Jeremy Ono
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Santa Jeremy Ono.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chau, KY., Manfioletti, G., Cheung-Chau, KW. et al. Derepression of HMGA2 Gene Expression in Retinoblastoma Is Associated with Cell Proliferation. Mol Med 9, 154–165 (2003). https://doi.org/10.1007/BF03402180

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF03402180

Keywords

Search

Navigation