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  • Oncogenomics
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Aberrant promoter methylation and silencing of the RASSF1A gene in pediatric tumors and cell lines

Oncogene volume 21, pages 4345–4349 (2002)Cite this article

Abstract

Aberrant promoter methylation of tumor suppressor genes has not been fully investigated in pediatric tumors. Therefore, we examined the methylation status of nine genes (p16INK4A, MGMT, GSTP1, RASSF1A, APC, DAPK, RARβ, CDH1 and CDH13) in 175 primary pediatric tumors and 23 tumor cell lines using methylation-specific PCR. We studied the major forms of pediatric tumors – Wilms' tumor, neuroblastoma, hepatoblastoma, medulloblastoma, rhabdomyosarcoma, osteosarcoma, Ewing's sarcoma, retinoblastoma and acute leukemia. The most frequently methylated gene in both primary tumors and cell lines was RASSF1A (40, 86%, respectively). However, the rates of RASSF1A methylation in individual tumor types varied from 0 to 88%. RASSF1A methylation was tumor specific and was absent in adjacent non-malignant tissues. Methylation of the other genes was relatively rare in tumors and non-malignant tissues (less than 5%). Neuroblastoma patients with methylation of RASSF1A were significantly older than patients without methylation (P=0.008). There was no relationship between methylation status and other clinico-pathologic parameters. We treated six cell lines lacking RASSF1A mRNA with 5-aza-2′deoxycytidine to examine the relationship between methylation and transcriptional silencing. In five of six cell lines, restoration of RASSF1A mRNA was confirmed by RT–PCR. Our findings indicate that aberrant promoter methylation of RASSF1A may contribute to the pathogenesis of many different forms of pediatric tumors.

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Abbreviations

TSG:

tumor suppressor gene

RASSF1A :

Ras association domain family 1, isoform A

MGMT :

O6-methylguanine DNA methyltransferase

GSTP1 :

glutathionne S-transferase P1

APC :

adenomatous polyposis coli

DAPK :

death-associated protein kinase

RARβ:

retinoic acid receptor-β

CDH1 :

E-cadherin

CDH13 :

H-cadherin

MSP:

methylation-specific PCR

RT–PCR:

reverse transcription-PCR

5-aza-CdR:

5-aza-2′deoxycytidine

References

  • Astuti D, Agathanggelou A, Honorio S, Dallol A, Martinsson T, Kogner P, Cummins C, Neumann HP, Voutilainen R, Dahia P, Eng C, Maher ER, Latif F . 2001 Oncogene 20: 7573–7577

  • Baylin SB, Herman JG, Graff JR, Vertino PM, Issa JP . 1998 Adv. Cancer Res. 72: 141–196

  • Burbee DG, Forgacs E, Zochbauer-Muller S, Shivakumar L, Fong K, Gao B, Randle D, Kondo M, Virmani A, Bader S, Sekido Y, Latif F, Milchgrub S, Toyooka S, Gazdar AF, Lerman MI, Zabarovsky E, White M, Minna JD . 2001 J. Natl. Cancer Inst. 93: 691–699

  • Chen B, Dias P, Jenkins III JJ, Savell VH, Parham DM . 1998 Am. J. Pathol. 152: 1071–1079

  • Dammann R, Li C, Yoon JH, Chin PL, Bates S, Pfeifer GP . 2000 Nat. Genet. 25: 315–319

  • Dammann R, Yang G, Pfeifer GP . 2001 Cancer Res. 61: 3105–3109

  • Davidoff AM, Hill DA . 2001 Semin. Pediatr. Surg. 10: 106–118

  • Dreijerink K, Braga E, Kuzmin I, Geil L, Duh FM, Angeloni D, Zbar B, Lerman MI, Stanbridge EJ, Minna JD, Protopopov A, Li J, Kashuba V, Klein G, Zabarovsky ER . 2001 Proc. Natl. Acad. Sci. USA 98: 7504–7509

  • Esteller M, Corn PG, Baylin SB, Herman JG . 2001 Cancer Res. 61: 3225–3229

  • Esteller M, Corn PG, Urena JM, Gabrielson E, Baylin SB, Herman JG . 1998 Cancer Res. 58: 4515–4518

  • Esteller M, Hamilton SR, Burger PC, Baylin SB, Herman JG . 1999a Cancer Res. 59: 793–797

  • Esteller M, Sanchez-Cespedes M, Rosell R, Sidransky D, Baylin SB, Herman JG . 1999b Cancer Res. 59: 67–70

  • Fruhwald MC, O'Dorisio MS, Dai Z, Rush LJ, Krahe R, Smiraglia DJ, Pietsch T, Elsea SH, Plass C . 2001a Genes Chrom. Cancer 30: 38–47

  • Fruhwald MC, O'Dorisio MS, Dai Z, Tanner SM, Balster DA, Gao X, Wright FA, Plass C . 2001b Oncogene 20: 5033–5042

  • Graff JR, Herman JG, Myohanen S, Baylin SB, Vertino PM . 1997 J. Biol. Chem. 272: 22322–22329

  • Herman JG, Graff JR, Myohanen S, Nelkin BD, Baylin SB . 1996 Proc. Natl. Acad. Sci. USA 93: 9821–9826

  • Loeb LA . 2001 Cancer Res. 61: 3230–3239

  • Mares J, Kriz V, Weinhausel A, Vodickova S, Kodet R, Haas OA, Sedlacek Z, Goetz P . 2001 Cancer Lett. 166: 165–171

  • Maruyama R, Toyooka S, Toyooka KO, Harada K, Virmani AK, Zöchbauer-Muller S, Farinas AJ, Vakar-Lopez F, Minna JD, Sagalowsky A, Czerniak B, Gazdar AF . 2001 Cancer Res. 61: 8659–8663

  • Maruyama R, Toyooka S, Toyooka KO, Virmani AK, Zöchbauer-Muller S, Farinas AJ, Minna JD, McConnel J, Frenkel EP, Gazdar AF . 2002 Clin. Cancer Res. 8: 514–519

  • Mollerup S, Ryberg D, Hewer A, Phillips DH, Haugen A . 1999 Cancer Res. 59: 3317–3320

  • Morrissey C, Martinez A, Zatyka M, Agathanggelou A, Honorio S, Astuti D, Morgan NV, Moch H, Richards FM, Kishida T, Yao M, Schraml P, Latif F, Maher ER . 2001 Cancer Res. 61: 7277–7281

  • Nakamura M, Sugita K, Inukai T, Goi K, Iijima K, Tezuka T, Kojika S, Shiraishi K, Miyamoto N, Karakida N, Kagami K, Koyama T, Mori T, Nakazawa S . 1999 Leukemia 13: 884–890

  • Ohtani-Fujita N, Dryja TP, Rapaport JM, Fujita T, Matsumura S, Ozasa K, Watanabe Y, Hayashi K, Maeda K, Kinoshita S, Matsumura T, Ohnishi Y, Hotta Y, Takahashi R, Kato MV, Ishizaki K, Sasaki MS, Horsthemke B, Minoda K, Sakai T . 1997 Cancer Genet. Cytogenet. 98: 43–49

  • Sato M, Mori Y, Sakurada A, Fujimura S, Horii A . 1998 Hum. Genet. 103: 96–101

  • Takita J, Yang HW, Bessho F, Hanada R, Yamamoto K, Kidd V, Teitz T, Wei T, Hayashi Y . 2000 Med. Pediatr. Oncol. 35: 541–543

  • Teitz T, Wei T, Valentine MB, Vanin EF, Grenet J, Valentine VA, Behm FG, Look AT, Lahti JM, Kidd VJ . 2000 Nat. Med. 6: 529–535

  • Toyooka KO, Toyooka S, Virmani AK, Sathyanarayana UG, Euhus DM, Gilcrease M, Minna JD, Gazdar AF . 2001a Cancer Res. 61: 4556–4560

  • Toyooka S, Pass HI, Shivapurkar N, Fukuyama Y, Maruyama R, Toyooka KO, Gilcrease M, Farinas A, Minna JD, Gazdar AF . 2001b Cancer Res. 61: 5727–5730

  • Tsuchiya T, Tamura G, Sato K, Endoh Y, Sakata K, Jin Z, Motoyama T, Usuba O, Kimura W, Nishizuka S, Wilson KT, James SP, Yin J, Fleisher AS, Zou T, Silverberg SG, Kong D, Meltzer SJ . 2000 Oncogene 19: 3642–3646

  • Virmani AK, Rathi A, Zochbauer-Muller S, Sacchi N, Fukuyama Y, Bryant D, Maitra A, Heda S, Fong KM, Thunnissen F, Minna JD, Gazdar AF . 2000 J. Natl. Cancer Inst. 92: 1303–1307

  • Wong IH, Ng MH, Huang DP, Lee JC . 2000 Blood 95: 1942–1949

  • Yoon JH, Dammann R, Pfeifer GP . 2001 Int. J. Cancer 94: 212–217

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Acknowledgements

This work was supported by grant U01CA8497102 from the Early Detection Research Network, National Cancer Institute, Bethesda, MD, USA.

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Authors and Affiliations

  1. Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, 75390, Texas, TX, USA

    Kenichi Harada, Shinichi Toyooka, Riichiroh Maruyama, Kiyomi O Toyooka, Gail E Tomlinson, John D Minna & Adi F Gazdar

  2. Department of Pathology, University of Texas Southwestern Medical Center, Dallas, 75390, Texas, TX, USA

    Anirban Maitra & Adi F Gazdar

  3. Department of Pediatrics, University of Texas Southwestern Medical Center, Dallas, 75390, Texas, TX, USA

    Gail E Tomlinson

  4. Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, 75390, Texas, TX, USA

    John D Minna

  5. Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, 75390, Texas, TX, USA

    John D Minna

  6. Department of Pathology, Children's Medical Center, Dallas, 75235, Texas, TX, USA

    Charles F Timmons

  7. Department of Ophthalmology, University of Siena, Siena, 53100, Italy

    Domenico Mastrangelo

  8. American Type Culture Collection, Manassas, 20110, Virginia, VA, USA

    Robert J Hay

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  1. Kenichi Harada
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  2. Shinichi Toyooka
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  11. Adi F Gazdar
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Correspondence to Adi F Gazdar.

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Harada, K., Toyooka, S., Maitra, A. et al. Aberrant promoter methylation and silencing of the RASSF1A gene in pediatric tumors and cell lines. Oncogene 21, 4345–4349 (2002). https://doi.org/10.1038/sj.onc.1205446

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