title: Lack of presence of the human cytomegalovirus in human glioblastoma.
abstract: Recent reports have indicated human cytomegalovirus (HCMV) to be associated with human glioblastoma carcinogenesis. In established examples of viral carcinogenesis, viral DNA and one or more of its products have been detected in most tumor cells of biopsies in the majority of cases. To test whether HCMV is associated with human glioblastoma based on this criterion, we measured the number of viral DNA molecules per cell in both frozen and paraffin-embedded tumor biopsies from 58 patients using real-time quantitative PCR (QPCR). Immunohistochemical and fluorescence in situ hybridization (FISH) to detect HCMV proteins and genome was performed in 10 cases using formalin-fixed paraffin-embedded glioblastoma tissues. Southern blotting using DNA extracted from four glioblastoma cell lines together with immunoblotting using the four cell lines and five glioblastoma tissue samples were also performed. We further confirmed the immunoblot bands using liquid chromatography-tandem mass spectrometry assay. As a result, HCMV DNA was not detected in the tumor cells from any of the glioblastoma cases by QPCR detecting two different HCMV genes, in clear contrast to samples from patients with HCMV infection. Southern blotting and immunoblotting of cell lines and FISH using paraffin sections were all negative. However, immunoblotting and immunohistochemistry using tissue samples were partly positive, but HCMV proteins were not detected by proteomic analysis, suggesting false positivity of the analyses. As our QPCR analysis could detect 10 copies of HCMV DNA mixed with DNA extracted from 10(4) HCMV-negative cells, we conclude that HCMV is not persistent, at least in the tumor cells, of developed human glioblastoma.
authors: 1] Department of Experimental Pathology and Tumor Biology, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan [2] Department of Pathology and Biological Responses, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors: Yoriko
authors: Y
authors: Yamashita
authors: Department of Pediatrics, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors: Yoshinori
authors: Y
authors: Ito
authors: Department of Virology and Preventive Medicine, Gunma University Graduate School of Medicine, Maebashi, Gunma, Japan.
authors: Hiroki
authors: H
authors: Isomura
authors: Department of Pathology and Biological Responses, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors: Naoaki
authors: N
authors: Takemura
authors: Department of School Nursing and Health Education, Aichi University of Education, Kariya, Aichi, Japan.
authors: Akira
authors: A
authors: Okamoto
authors: Department of Neurosurgery, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors: Kazuya
authors: K
authors: Motomura
authors: Department of Neurosurgery, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors: Takashi
authors: T
authors: Tsujiuchi
authors: Department of Neurosurgery, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors: Atsushi
authors: A
authors: Natsume
authors: Department of Neurosurgery, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors: Toshihiko
authors: T
authors: Wakabayashi
authors: Department of Pathology and Biological Responses, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors: Shinya
authors: S
authors: Toyokuni
authors: Division of Virology, Aichi Cancer Center Research Institute, Nagoya, Japan.
authors: Tatsuya
authors: T
authors: Tsurumi
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001932:Brain Neoplasms / Q000473:pathology / Q000821:virology*; D002648:Child; D003587:Cytomegalovirus / Q000302:isolation & purification*; D004279:DNA, Viral / Q000032:analysis*; D005260:Female; D005909:Glioblastoma / Q000473:pathology / Q000821:virology*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D008875:Middle Aged; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D004279:DNA, Viral