Human Genetics

, Volume 125, Issue 2, pp 189–198 | Cite as

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

  • Susmita Ghosh
  • Amlan Ghosh
  • Guru Prasad Maiti
  • Neyaz Alam
  • Anup Roy
  • Susanta Roychoudhury
  • Chinmay Kumar Panda
Original Investigation
First Online:
Received:
Accepted:

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.

Keywords

Molecular Alteration SCC084 Cell Dysplastic Lesion Oral Cavity Cancer Oral Cancer Cell Line 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Notes

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.

Conflict of interest statement

None.

Supplementary material

439_2008_610_MOESM1_ESM.doc (172 kb)
Supplementary material 1 (DOC 172 kb)
439_2008_610_MOESM2_ESM.pdf (103 kb)
Supplementary material 1 (PDF 103 kb)

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Susmita Ghosh
    • 1
  • Amlan Ghosh
    • 1
  • Guru Prasad Maiti
    • 1
  • Neyaz Alam
    • 2
  • Anup Roy
    • 3
  • Susanta Roychoudhury
    • 4
  • Chinmay Kumar Panda
    • 1
  1. 1.Department of Oncogene RegulationChittaranjan National Cancer InstituteKolkataIndia
  2. 2.Department of SurgeryChittaranjan National Cancer InstituteKolkataIndia
  3. 3.Calcutta Medical College and HospitalKolkataIndia
  4. 4.Molecular and Human Genetics and Genomic DivisionIndian Institute of Chemical BiologyKolkataIndia

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