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Alterations of ROBO1/DUTT1 and ROBO2 loci in early dysplastic lesions of head and neck: clinical and prognostic implications

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Abstract

Deletion of chromosomal 3p12.3 was suggested to be associated with dysplastic lesions of head and neck. This region harbors two candidate tumor suppressors ROBO1/DUTT1, ROBO2 and two non-coding RNAs (ncRNAs) located at intron 2 of ROBO1/DUTT1. Aim of this study is to understand the role of these genes in development of head and neck squamous cell carcinoma. A collection of 72 dysplastic lesions and 116 HNSCC samples and two oral cancer cell lines were analyzed for ROBO1/DUTT1 and ROBO2 deletion and promoter methylation. ROBO1/DUTT1, ROBO2 and two ncRNAs mRNA expression were analyzed by Q-PCR. Immunohistochemical analysis of ROBO1/DUTT1 and ROBO2 was performed. Alterations of these genes were correlated with different clinicopathological parameters. High frequency of molecular alterations (deletion/methylation) was seen in ROBO1/DUTT1 than ROBO2. In mild dysplastic lesions both of these genes showed high molecular alterations and remained more or less constant in subsequent stages. Q-PCR analysis showed reduced expression of these genes and the two ncRNAs. In vitro demethylation experiment by 5-aza-dC showed upregulation of ROBO1/DUTT1 and ROBO2 while the expression of the ncRNAs remained unchanged. Immunohistochemical analysis of ROBO1/DUTT1 and ROBO2 showed concordance with their mRNA expression and molecular alterations. Poor patients’ outcome was predicted in the cases with alteration of ROBO1/DUTT1 along with tobacco addiction and nodal involvement. Our data suggests (a) ROBO1/DUTT1 and the ncRNAs are transcribed from different promoters, and (b) inactivation of ROBO1/DUTT1 could be used as molecular signature for early detection and prognosis of the head and neck cancer.

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Acknowledgments

We are thankful to the Directors, Chittaranjan National Cancer Institute and Cancer Center & Welfare Home, Kolkata, India. We are also grateful to Prof. H. Z. Hausen and Prof. E. M. de Villiers for their generous gift of HPV-16/18 plasmids. We also thank Prof. Susanne M. Gollin for UPCI: SCC084 cell line. Sources of support: Financial support for this work was provided by grants [SR/SO/BB-22/2003 dt. 02.11.04] from DST, and [BT/PR/5524/Med/14/649/2004 of dt. 29.11.2005] from DBT, Govt. of India to Dr. C. K. Panda and Dr. S. Roychoudhury; and UGC-NET Fellowship grant [F.2-3/2000 (SA-I)] to Mrs. S. Ghosh, CSIR-NET Fellowship grant [F.2-3/2000 (SA-I)] to Mr. A.Ghosh.

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Authors and Affiliations

  1. Department of Oncogene Regulation, Chittaranjan National Cancer Institute, 37, S.P. Mukherjee Road, Kolkata, 700026, India

    Susmita Ghosh, Amlan Ghosh, Guru Prasad Maiti & Chinmay Kumar Panda

  2. Department of Surgery, Chittaranjan National Cancer Institute, 37, S.P. Mukherjee Road, Kolkata, 700026, India

    Neyaz Alam

  3. Calcutta Medical College and Hospital, 88 College Street, Kolkata, 700073, India

    Anup Roy

  4. Molecular and Human Genetics and Genomic Division, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Kolkata, 700032, India

    Susanta Roychoudhury

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  1. Susmita Ghosh
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  2. Amlan Ghosh
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Correspondence to Chinmay Kumar Panda.

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Ghosh, S., Ghosh, A., Maiti, G.P. et al. Alterations of ROBO1/DUTT1 and ROBO2 loci in early dysplastic lesions of head and neck: clinical and prognostic implications. Hum Genet 125, 189–198 (2009). https://doi.org/10.1007/s00439-008-0610-9

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  • DOI: https://doi.org/10.1007/s00439-008-0610-9

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