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  • Original Paper
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Tumour specific promoter region methylation of the human homologue of the Drosophila Roundabout gene DUTT1 (ROBO1) in human cancers

Oncogene volume 21, pages 3020–3028 (2002)Cite this article

Abstract

The human homologue of the Drosophila Roundabout gene DUTT1 (Deleted in U Twenty Twenty) or ROBO1 (Locus Link ID 6091), a member of the NCAM family of receptors, was recently cloned from the lung cancer tumour suppressor gene region 2 (LCTSGR2 or U2020 region) at 3p12. DUTT1 maps within a region of overlapping homozygous deletions characterized in both small cell lung cancer lines (SCLC) and in a breast cancer line. In this report we (a) defined the genomic organization of the DUTT1 gene, (b) performed mutation and expression analysis of DUTT1 in lung, breast and kidney cancers, (c) identified tumour specific promoter region methylation of DUTT1 in human cancers. The gene was found to contain 29 exons and spans at least 240 kb of genomic sequence. The 5′ region contains a CpG island, and the poly(A)+ tail has an atypical 5′-GATAAA-3′ signal. We analysed DUTT1 for mutations in lung, breast and kidney cancers, no inactivating mutations were detected by PCR–SSCP. However, seven germline missense changes were found and characterized. DUTT1 expression was not detectable in one out of 18 breast tumour lines analysed by RT–PCR. Bisulfite sequencing of the promoter region of DUTT1 gene in the HTB-19 breast tumour cell line (not expressing DUTT1) showed complete hypermethylation of CpG sites within the promoter region of the DUTT1 gene (−244 to +27 relative to the translation start site). The expression of DUTT1 gene was reactivated in HTB-19 after treatment with the demethylating agent 5-aza-2′-deoxycytidine. The same region was also found to be hypermethylated in six out of 32 (19%) primary invasive breast carcinomas and eight out of 44 (18%) primary clear cell renal cell carcinomas (CC–RCC) and in one out of 26 (4%) primary NSCLC tumours. Furthermore 80% of breast and 75% of CC–RCC tumours showing DUTT1 methylation had allelic losses for 3p12 markers hence obeying Knudson's two hit hypothesis. Our findings suggest that DUTT1 warrants further analysis as a candidate for the tumour suppressor gene (TSG) at 3p12, a region defined by hemi and homozygous deletions and functional analysis.

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References

  • Agathanggelou A, Honorio S, Macartney DP, Martinez A, Dallol A, Rader J, Fullwood P, Chauhan A, Walker R, Shaw JA, Hosoe S, Lerman MI, Minna JD, Maher ER, Latif F . 2001 Oncogene 20: 1509–1518

  • Angeloni D, Wei MH, Timmer T, Van Der Veen AY, Buys CH, Lerman M . 2000 Cancer Genetics and Tumor Suppressor Genes Meeting Cold Spring Harbor Laboratories abstract

    Google Scholar 

  • Bashaw GJ, Kidd T, Murray D, Pawson T, Goodman CS . 2000 Cell 101: 703–715

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

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

  • Clifford SC, Prowse AH, Affara NA, Buys CH, Maher ER . 1998 Genes Chrom. Cancer 22: 200–209

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

  • Drabkin HA, Mendez MJ, Rabbitts PH, Varkony T, Bergh J, Schlessinger J, Erickson P, Gemmill RM . 1992 Genes Chrom. Cancer 5: 67–74

  • Elston CW, Ellis IO . 1991 Histopathology 19: 403–410

  • Foster K, Osborne RJ, Huddart RA, Affara NA, Ferguson-Smith MA, Maher ER . 1995 Eur. J. Cancer 31A: 2392–2395

  • Gnarra JR, Tory K, Weng Y, Schmidt L, Wei MH, Li H, Latif F, Liu S, Chen F, Duh FM et al . 1994 Nat. Genet. 7: 85–90

  • Herman JG, Latif F, Weng Y, Lerman MI, Zbar B, Liu S, Samid D, Duan DS, Gnarra JR, Linehan WM, Baylin SB . 1994 Proc. Natl. Acad. Sci. USA 91: 9700–9704

  • Huebner K, Garrison PN, Barnes LD, Croce CM . 1998 Annu. Rev. Genet. 32: 7–31

  • Kaelin Jr WG, Maher ER . 1998 Trends Genet. 14: 423–426

  • Keino-Masu K, Masu M, Hinck L, Leonardo ED, Chan SS, Culotti JG, Tessier-Lavigne M . 1996 Cell 87: 175–185

  • Kidd T, Bland KS, Goodman CS . 1999 Cell 96: 785–794

  • Kidd T, Brose K, Mitchell KJ, Fetter RD, Tessier-Lavigne M, Goodman CS, Tear G . 1998 Cell 92: 205–215

  • Kok K, Naylor SL, Buys CH . 1997 Adv. Cancer Res. 71: 27–92

  • Latif F, Tory K, Gnarra J, Yao M, Duh FM, Orcutt ML, Stackhouse T, Kuzmin I, Modi W, Geil L, Schmidt L, Zhou F, Li H, Wei MH, Chen F, Glenn G, Choyke P, Walther MM, Weng Y, Duan DSR, Dean M, Glavac D, Richards FM, Crossey PA, Ferguson-Smith MA, Paslier D, Chumakov I, Cohen D, Chinault AC, Maher ER, Linehan WM, Zbar B, Lerman MI . 1993 Science 260: 1317–1320

  • Latif F, Tory K, Modi WS, Graziano SL, Gamble G, Douglas J, Heppell-Parton AC, Rabbitts PH, Zbar B, Lerman MI . 1992 Genes Chrom. Cancer 5: 119–127

  • Lerman MI, Minna JD, for the International Lung Cancer Chromosome 3p21.3 Tumor Suppressor Gene Consortium. 2000 Cancer Res. 60: 6116–6133

  • Lott ST, Lovell M, Naylor SL, Killary AM . 1998 Cancer Res. 58: 3533–3537

  • Lovell M, Lott ST, Wong P, El Naggar A, Tucker S, Killiary AM . 1999 Cancer Res. 59: 2182–2189

  • Lubinski J, Hadaczek P, Podolski J, Toloczko A, Sikorski A, McCue P, Druck T, Huebner K . 1994 Cancer Res. 54: 3710–3713

  • Man S, Ellis IO, Sibbering M, Blamey RW, Brook JD . 1996 Cancer Res. 56: 5484–5489

  • Martinez A, Fullwood P, Kondo K, Kishida T, Yao M, Maher ER, Latif F . 2000 Mol. Pathol. 53: 137–144

  • Martinez A, Walker RA, Shaw JA, Dearing SJ, Maher ER, Latif F . 2001 Mol. Pathol. 54: 300–306

  • 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

  • Muller A, Homey B, Soto H, Ge N, Catron D, Buchanan ME, McClanahan T, Murphy E, Yuan W, Wagner SN, Barrera JL, Mohar A, Verastegui E, Zlotnik A . 2001 Nature 410: 50–56

  • Phelps RM, Johnson BE, Ihde DC, Gazdar AF, Carbone DP, McClintock PR, Linnoila RI, Matthews MJ, Bunn Jr PA, Carney D, Minna JD, Mulshine JL . 1996 J. Cell Biochem. 24: Suppl 32–91

  • Rabbitts P, Bergh J, Douglas J, Collins F, Waters J . 1990 Genes Chrom. Cancer 2: 231–238

  • Sato K, Tamura G, Tsuchiya T, Endoh Y, Usuba O, Kimura W, Motoyama T . 2001 Br. J. Cancer 85: 199–203

  • Shaw JA, Walsh T, Chappell SA, Carey N, Johnson K, Walker RA . 1996 Br. J. Cancer 73: 1393–1397

  • Shuin T, Kondo K, Torigoe S, Kishida T, Kubota Y, Hosaka M, Nagashima Y, Kitamura H, Latif F, Zbar B, Lerman MI, Yao M . 1994 Cancer Res. 54: 2852–2855

  • Stein E, Tessier-Lavigne M . 2001 Science 291: 1928–1938

  • Sundaresan V, Chung G, Heppell-Parton A, Xiong J, Grundy C, Roberts I, James L, Cahn A, Bench A, Douglas J, Minna JD, Sekido Y, Lerman MI, Latif F, Bergh J, Li H, Lowe N, Ogilvie D, Rabbitts P . 1998a Oncogene 17: 1723–1729

  • Sundaresan V, Roberts I, Bateman A, Bankier A, Sheppard M, Hobbs C, Xiong J, Minna JD, Latif F, Lerman MI, Rabbitts P . 1998b Mol. Cell. Neurosci. 11: 29–35

  • Thoenes W, Storkel S, Rumpelt HJ . 1986 Path. Res. Pract. 181: 125–143

  • Todd S, Franklin WA, Varella-Garcia M, Kennedy T, Hilliker Jr CE, Hahner L, Anderson M, Wiest JS, Drabkin HA, Gemmill RM . 1997 Cancer Res. 57: 1344–1352

  • Wistuba II, Behrens C, Milchgrub S, Syed S, Ahmadian M, Virmani AK, Kurvari V, Cunningham TH, Ashfaq R, Minna JD, Gazdar AF . 1998 Clin. Cancer Res. 4: 2931–2938

  • Wistuba II, Behrens C, Virmani AK, Mele G, Milchgrub S, Girard L, Fondon III JW, Garner HR, McKay B, Latif L, Lerman MI, Lam S, Gazdar AF, Minna JD . 2000 Cancer Res. 60: 1949–1960

  • Wu JY, Feng L, Park H-T, Havlioglu N, Tang LH, Bacon KB, Jiang Z-H, Zhan X-C, Rao Y . 2001 Nature 410: 948–951

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Acknowledgements

This work is supported in part by Cancer Research UK, Breast Cancer Campaign, Lung Cancer SPORE P50 CA70907, and the G Harold and Leila Y Mathers Charitable Foundation. A Martinez is supported by the University of Antioquia, Medellin, Colombia, and a fellowship from COLCIENCIAS.

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Author notes

  1. Eva Forgacs and Alonso Martinez: E Forgacs and A Martinez contributed equally to this study

Authors and Affiliations

  1. Department of Paediatrics and Child Health, Section of Medical and Molecular Genetics, University of Birmingham, The Medical School, Edgbaston, Birmingham, B15 2TT, UK

    Ashraf Dallol, Alonso Martinez, Eamonn R Maher & Farida Latif

  2. Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas, USA

    Eva Forgacs, Yoshitaka Sekido & John D Minna

  3. Breast Cancer Research Unit, University of Leicester, Clinical Sciences, Glenfield Hospital, Leicester, UK

    Rosemary Walker

  4. Department of Urology, Yokohama City University School of Medicine, Yokohama, Japan

    Takeshi Kishida

  5. Department of Oncology, MRC Molecular Oncology Group, University of Cambridge, MRC Centre, Hills Road, Cambridge, CB2 2QH, UK

    Pamela Rabbitts

  6. Cancer Research UK Renal Molecular Oncology Research Group, University of Birmingham, The Medical School, Edgbaston, Birmingham, B15 2TT, UK

    Eamonn R Maher & Farida Latif

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  1. Ashraf Dallol
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  2. Eva Forgacs
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Correspondence to Farida Latif.

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Dallol, A., Forgacs, E., Martinez, A. et al. Tumour specific promoter region methylation of the human homologue of the Drosophila Roundabout gene DUTT1 (ROBO1) in human cancers. Oncogene 21, 3020–3028 (2002). https://doi.org/10.1038/sj.onc.1205421

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