title: Evaluation of Ki-67 reactivity in neuroblastoma using paraffin embedded tissue.
abstract: OBJECTIVE
  To examine the pattern of reactivity of Ki-67 in neuroblastoma and correlate this with a) clinical prognostic criteria and b) cell cycle statistics (using flow cytometry).
  
  
  METHODS
  Four micron sections of paraffin embedded (PE) tissue from 55 patients (25 pre chemotherapy and 30 post) were placed on to aminosialinised slides, dewaxed and rehydrated. Slides were pretreated in a microwave oven, endogenous peroxidase activity blocked using 3% hydrogen peroxide and Ki-67 reactivity investigated using a streptavidin/biotin/peroxidase technique. DNA ploidy was also performed from an immediately adjacent section on the same block using a FACScan and Cellfit software.
  
  
  RESULTS
  Ki-67 reactivity was well defined and highly reproducible. Eighteen out of 30 post chemotherapy samples were totally negative, despite evidence of proliferation on flow cytometry and all subsequently died of disease. As interpretation post chemotherapy was therefore deemed unreliable, this group was excluded from analysis. Reactivity in pretreatment samples ranged from 0% to 67%; staining was restricted to the nucleus with a distinct pattern noted in the nucleolus. Ki-67 positivity was lower in aneuploid compared with diploid tumours (mean 26% vs 36%, NS). Among diploid tumours, a lower percentage positivity was noted in those patients with better clinical prognostic parameters. Correlation however between Ki-67 and SG2M phases of cell cycle was poor (RS = 0.39, NS).
  
  
  CONCLUSIONS
  Assessment of proliferation in neuroblastoma by Ki-67 reactivity in paraffin embedded tissue is reliable in pretreatment samples and can be incorporated into routine immunohistochemical evaluation. Larger multicentre studies are required to further evaluate Ki-67 reactivity as a prognostic indicator.
authors.affiliation: Department of Pathology, Mater Misericordiae Hospital, Dublin, Ireland.
authors.forename: D
authors.initials: D
authors.lastname: Graham
authors.forename: H
authors.initials: H
authors.lastname: Magee
authors.forename: B
authors.initials: B
authors.lastname: Kierce
authors.forename: R
authors.initials: R
authors.lastname: Ball
authors.forename: P
authors.initials: P
authors.lastname: Dervan
authors.forename: A
authors.initials: A
authors.lastname: O'Meara
mesh_terms: D014408:Biomarkers, Tumor / Q000032:analysis*; D002453:Cell Cycle; D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D019394:Ki-67 Antigen; D008297:Male; D009363:Neoplasm Proteins / Q000032:analysis*; D009447:Neuroblastoma / Q000737:chemistry / Q000401:mortality / Q000473:pathology*; D009687:Nuclear Proteins / Q000032:analysis*; D016612:Paraffin Embedding; D011379:Prognosis
chemical_list: D014408:Biomarkers, Tumor; D019394:Ki-67 Antigen; D009363:Neoplasm Proteins; D009687:Nuclear Proteins