title: Involvement of PTEN promoter methylation in cerebral cavernous malformations.
abstract: OBJECTIVE
  Cerebral cavernous malformations (CCMs) are prevalent cerebral vascular lesions involving aberrant angiogenesis. However, the underlying mechanism is poorly understood. Phosphatase and tension homolog deleted on chromosome 10 (PTEN), a tumor suppressor, is frequently deficient in various pathologies due to mutation or epigenetic alterations. PTEN promoter hypermethylation is a major epigenetic silencing mechanism leading to activation of angiogenesis in tumors. The present study aimed to investigate whether PTEN promoter methylation was involved in CCMs.
  
  
  METHODS
  PTEN promoter methylation was detected in surgical specimens of CCMs (n=69) by methylation-specific polymerase chain reaction. The methylation status was correlated to the clinical manifestations and to PTEN expression, which was analyzed by both Western blot and immunohistochemistry. To investigate the endothelial proliferation and the potential signaling pathways affected by PTEN methylation, proliferating cell nuclear antigen as well as phosphor-Akt and phosphor-Erk1,2 were detected by immunofluorescence and Western blot, respectively, in CCM specimens.
  
  
  RESULTS
  Methylation-specific polymerase chain reaction revealed PTEN promoter methylation in 15.9% CCMs. Strikingly, 5 of 6 familial CCMs showed PTEN promoter methylation (83.3%), which was significantly higher than in sporadic cases (9.4%; P<0.001). In addition, PTEN promoter methylation appeared more frequently in multiple CCMs, including familial cases (46.7%), than that in single-lesioned CCMs (11.8%; P<0.05). Immunostaining and Western blot revealed a more significant PTEN downregulation in PTEN-methylated CCMs in comparison to PTEN-unmethylated CCMs. Reduced PTEN expression was inversely correlated to the expression of proliferating cell nuclear antigen and to the activation of Erk1,2, but not of Akt.
  
  
  CONCLUSIONS
  We reported here for the first time the involvement of PTEN promoter methylation in CCMs, particularly in familial CCMs, suggesting this epigenetic alteration as a potential pathomechanism of CCMs. The identification of Erk1,2 as triggered signaling in the lesions may be valuable for the development of effective therapy for this disease.
authors.affiliation: Department of Neurosurgery, University of Marburg, Marburg, Germany. yuan.zhu@uk-essen.de
authors.forename: Yuan
authors.initials: Y
authors.lastname: Zhu
authors.forename: Andreas
authors.initials: A
authors.lastname: Wloch
authors.forename: Qun
authors.initials: Q
authors.lastname: Wu
authors.forename: Christian
authors.initials: C
authors.lastname: Peters
authors.forename: Axel
authors.initials: A
authors.lastname: Pagenstecher
authors.forename: Helmut
authors.initials: H
authors.lastname: Bertalanffy
authors.forename: Ulrich
authors.initials: U
authors.lastname: Sure
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects; D002648:Child; D002675:Child, Preschool; D004247:DNA / Q000096:biosynthesis / Q000302:isolation & purification; D042783:Endothelial Cells / Q000187:drug effects; D048049:Extracellular Signal-Regulated MAP Kinases / Q000235:genetics / Q000378:metabolism; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D002538:Intracranial Arteriovenous Malformations / Q000235:genetics*; D008279:Magnetic Resonance Imaging; D008297:Male; D008745:Methylation; D008875:Middle Aged; D032389:Myocytes, Smooth Muscle / Q000187:drug effects; D009389:Neovascularization, Pathologic / Q000235:genetics; D051059:PTEN Phosphohydrolase / Q000235:genetics* / Q000502:physiology*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects; D013447:Sulfites / Q000378:metabolism; D055815:Young Adult
chemical_list: D013447:Sulfites; D004247:DNA; D051057:Proto-Oncogene Proteins c-akt; D048049:Extracellular Signal-Regulated MAP Kinases; D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human