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

Ashraf Dallol, Eva Forgacs, Alonso Martinez, Yoshitaka Sekido, Rosemary Walker, Takeshi Kishida, Pamela Rabbitts, Eamonn R. Maher, John D. Minna, Farida Latif

Research output: Contribution to journalArticle

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 turnout fines analysed by RT-PCR. Bisulfite sequencing of the promoter region of DUTT1 gene in the HTB-19 breast turnout 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 he 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.

Original languageEnglish (US)
Pages (from-to)3020-3028
Number of pages9
JournalOncogene
Volume21
Issue number19
DOIs
StatePublished - May 2 2002

Fingerprint

Genetic Promoter Regions
Carcinogens
Methylation
Drosophila
Tumor Suppressor Genes
Breast Neoplasms
Lung Neoplasms
Breast
Kidney Neoplasms
Genes
decitabine
Neoplasms
Renal Cell Carcinoma
Mutation
Neural Cell Adhesion Molecules
Single-Stranded Conformational Polymorphism
Polymerase Chain Reaction
CpG Islands
Loss of Heterozygosity
Small Cell Lung Carcinoma

Keywords

  • 3p12 tumour suppressor gene
  • DUTT1/ROBO1
  • Methylation

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Genetics

Cite this

Dallol, A., Forgacs, E., Martinez, A., Sekido, Y., Walker, R., Kishida, T., ... Latif, F. (2002). Tumour specific promoter region methylation of the human homologue of the Drosophila Roundabout gene DUTT1 (ROBO1) in human cancers. Oncogene, 21(19), 3020-3028. https://doi.org/10.1038/sj/onc/1205421

Tumour specific promoter region methylation of the human homologue of the Drosophila Roundabout gene DUTT1 (ROBO1) in human cancers. / Dallol, Ashraf; Forgacs, Eva; Martinez, Alonso; Sekido, Yoshitaka; Walker, Rosemary; Kishida, Takeshi; Rabbitts, Pamela; Maher, Eamonn R.; Minna, John D.; Latif, Farida.

In: Oncogene, Vol. 21, No. 19, 02.05.2002, p. 3020-3028.

Research output: Contribution to journalArticle

Dallol, A, Forgacs, E, Martinez, A, Sekido, Y, Walker, R, Kishida, T, Rabbitts, P, Maher, ER, Minna, JD & Latif, F 2002, 'Tumour specific promoter region methylation of the human homologue of the Drosophila Roundabout gene DUTT1 (ROBO1) in human cancers', Oncogene, vol. 21, no. 19, pp. 3020-3028. https://doi.org/10.1038/sj/onc/1205421
Dallol, Ashraf ; Forgacs, Eva ; Martinez, Alonso ; Sekido, Yoshitaka ; Walker, Rosemary ; Kishida, Takeshi ; Rabbitts, Pamela ; Maher, Eamonn R. ; Minna, John D. ; Latif, Farida. / Tumour specific promoter region methylation of the human homologue of the Drosophila Roundabout gene DUTT1 (ROBO1) in human cancers. In: Oncogene. 2002 ; Vol. 21, No. 19. pp. 3020-3028.
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