title: Frequent HRK inactivation associated with low apoptotic index in secondary glioblastomas.
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 p16(INK4a) and p14(ARF). 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.
authors.affiliation: Department of Pathology, Nara Medical University School of Medicine, 840 Shijo-cho, Kashihara, 634-8521, Nara, Japan.
authors.forename: Mitsutoshi
authors.initials: M
authors.lastname: Nakamura
authors.forename: Eiwa
authors.initials: E
authors.lastname: Ishida
authors.forename: Keiji
authors.initials: K
authors.lastname: Shimada
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Nakase
authors.forename: Toshisuke
authors.initials: T
authors.lastname: Sakaki
authors.forename: Noboru
authors.initials: N
authors.lastname: Konishi
mesh_terms: D017209:Apoptosis / Q000502:physiology*; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism*; D002892:Chromosomes, Human, Pair 22; D015331:Cohort Studies; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology*; D005909:Glioblastoma / Q000145:classification / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D006801:Humans; D020287:In Situ Nick-End Labeling / Q000379:methods; D019656:Loss of Heterozygosity / Q000502:physiology; D008297:Male; D008745:Methylation; D012333:RNA, Messenger / Q000378:metabolism; D012189:Retrospective Studies; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
chemical_list: D051017:Apoptosis Regulatory Proteins; C484586:HRK protein, human; D012333:RNA, Messenger