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Frequent HRK inactivation associated with low apoptotic index in secondary glioblastomas

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Abstract

To detect and identify the genetic alterations and methylation status of the HRK gene in human glioblastomas, we analyzed a cohort of astrocytic tumors for hypermethylation, loss of heterozygosity on 12q13.1, and gene expression. Our study examined a series of 36 diffuse low-grade astrocytomas, 32 anaplastic astrocytomas, 64 primary glioblastomas, and 28 secondary glioblastomas that had evolved from either 24 low-grade diffuse astrocytomas or 4 anaplastic astrocytomas. The region around the HRK transcription start site was methylated in 19% of diffuse astrocytomas, in 22% of anaplastic astrocytomas, in 27% of primary glioblastomas, and in 43% of secondary glioblastomas. HRK expression was significantly reduced in 61% of secondary glioblastomas as compared to other types of tumors, and aberrant methylation was closely associated with loss of expression. Reverse transcription-PCR analysis also demonstrated a clear agreement between reduced HRK protein levels and low or absent HRK transcripts. Lack of HRK immunoreactivity was significantly correlated with a low apoptotic index, whereas a strong association between methylation status and apoptosis was found only in secondary glioblastomas. Abnormal methylation of HRK was detected in astrocytic tumors concurrent with methylation of multiple genes, including p16INK4a and p14ARF. Interestingly, these epigenetic changes in secondary glioblastoma were further associated with wild-type p53. Our findings suggest that HRK is inactivated mainly by aberrant DNA methylation in astrocytic tumors and that reduced HRK expression contributes to the loss of apoptotic control in high-grade tumors. Reduced expression of HRK may serve as one important molecular mechanism in progression to secondary glioblastoma.

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Acknowledgements

This work was supported by a Grant-in-Aid for Scientific Research (C) from the Ministry of Education, Culture, Sports, Science and Technology of Japan (no. 17590312).

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Authors and Affiliations

  1. Department of Pathology, Nara Medical University School of Medicine, 840 Shijo-cho, Kashihara, 634–8521, Nara, Japan

    Mitsutoshi Nakamura, Eiwa Ishida, Keiji Shimada & Noboru Konishi

  2. Department of Neurosurgery, Nara Medical University School of Medicine, 634–8521, Nara, Japan

    Hiroyuki Nakase & Toshisuke Sakaki

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  1. Mitsutoshi Nakamura
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  2. Eiwa Ishida
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  3. Keiji Shimada
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  4. Hiroyuki Nakase
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  5. Toshisuke Sakaki
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  6. Noboru Konishi
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Correspondence to Noboru Konishi.

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Nakamura, M., Ishida, E., Shimada, K. et al. Frequent HRK inactivation associated with low apoptotic index in secondary glioblastomas. Acta Neuropathol 110, 402–410 (2005). https://doi.org/10.1007/s00401-005-1065-x

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  • DOI: https://doi.org/10.1007/s00401-005-1065-x

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