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Altered structure and deregulated expression of the tumor suppressor gene retinoblastoma (RB1) in human brain tumors

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Abstract

A total of 40 human brain tumor samples were analyzed for tumor-specific alterations at the RB1 gene locus. Gliomas were more prevalent in younger males and meningiomas in older females. Southern blot analysis revealed loss of heterozygosity (LOH) at the intron 1 locus of RB1 gene in 19.4% of informative cases and this is the first report showing LOH at this locus in human brain tumors. Levels of RB1 mRNA and protein, pRb, and the percentage of hyperphosphorylated form of pRb were also analyzed in these tumors. Normal human fibroblast cell line WI38 was used as control in northern and western analysis. Normal sized RB1 mRNA and protein were present in all the tumor samples. Majority of the gliomas had 2.0-fold or higher levels of RB1 mRNA and most meningiomas had less than 2.0-fold of RB1 mRNA compared to control WI38 cells. The total pRb levels were 2.0-fold or higher in all the tumor samples compared to control. More than 50% of pRb existed in hyperphosphorylated form in all gliomas except two. However, six out of 13 meningiomas had less than 50% of total pRb in the hyperphosphorylated form. These results indicate that the increased percentage of hyperphosphorylated form of pRb in gliomas could provide growth advantage to these tumors. Presence of LOH at the RB1 gene locus and the increased levels of RB1 RNA and protein and increased percentage of hyperphosphorylated form of pRb are indicative of an overall deregulation of pRb pathway in human brain tumors.

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Acknowledgments

We Thank Dr. T.P. Dryja Department of Ophthalmology, Harvard Medical School for the plasmids p123M1.8 and p4.95BT. We thank Dr. B.A. Chandramouli, Dr. S. Sampath and Dr. B. Indira Devi, Department of Neurosurgery, NIMHANS, for providing some of the surgical tumor tissues. We thank Dr. D.K. Subbakrishna, Department of Biostatistics, NIMHANS, for statistical analysis of the data. We thank Dr. A.H.P.P. Dayalan, Department of Human Genetics, NIMHANS, for his help with some of the experiments. JM and RK are recipients of Senior Research Fellowship from University Grant Commission, Government of India. The infrastructure was provided by the Department of Biotechnology (DBT), Government of India (grant awarded to RG, project No. BT/PRO 703/09/133/97) and by Life Sciences Research Board (LSRB), Government of India (grant awarded to RG, project No. DBA/48222/c/RD-81). This work is toward the partial fulfillment of Ph.D. degree for JM.

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Authors and Affiliations

  1. Department of Human Genetics, National Institute of Mental Health and Neurosciences (NIMHANS), Bangalore, 560 029, India

    J. Mathivanan, K. Rohini & Rajalakshmi Gope

  2. Department of Biotechnology, City College, Bangalore, 560 070, India

    Mohan L. Gope

  3. Department of Neurosurgery, NIMHANS, Bangalore, 560 029, India

    B. Anandh

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  1. J. Mathivanan
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Correspondence to Rajalakshmi Gope.

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Mathivanan, J., Rohini, K., Gope, M.L. et al. Altered structure and deregulated expression of the tumor suppressor gene retinoblastoma (RB1) in human brain tumors. Mol Cell Biochem 302, 67–77 (2007). https://doi.org/10.1007/s11010-007-9428-3

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