title: Clinical relevance of DNA ploidy and proliferative activity in childhood rhabdomyosarcoma: a retrospective analysis of patients enrolled onto the Italian Cooperative Rhabdomyosarcoma Study RMS88.
abstract: OBJECTIVE
  Evaluation of the possible clinical relevance of DNA ploidy and proliferative activity assessed as S-phase fraction (SPF) in childhood rhabdomyosarcoma (RMS).
  
  
  METHODS
  We conducted a retrospective study on 59 RMS patients enrolled onto the ICS-RMS88 protocol (seven botryoid, 35 embryonal, and 17 alveolar RMS), for which formalin-fixed paraffin-embedded (FFPE) tissue was available. Nuclear suspensions for cytometric investigation were obtained using a mechanical disaggregation. Tumors were distinguished according to their DNA index (DI) value as follows: diploid (0.9 < DI < 1.1), hyperdiploid (1.1 < or = DI < 1.8 or DI > or = 2.2), and tetraploid (1.8 < or = DI < 2.2); for analysis of SPF, a cutoff value of 14% was used.
  
  
  RESULTS
  DNA histograms were diploid in 19 (33%) cases, hyperdiploid in 29 (49%), and tetraploid in 10 (32%). One patient showed both a hyperdiploid and a tetraploid peak. The 5-year overall survival (OS) rate by ploidy status was 73% in hyperdiploid patients as compared with 33% and 25% in diploid and tetraploid patients, respectively (P = .0012). A striking difference emerged when the 5-year OS for the combined diploid and tetraploid RMS groups was compared with survival of the hyperdiploid RMS group: 30% versus 73%, respectively (P = .0006). In addition, the SPF was prognostically relevant: 5-year OS by SPF less than or greater than 14% was 70% and 36%, respectively (P = .009). Multivariate analysis confirmed the importance of DNA content (P = .0006) and SPF (P = .034) in predicting survival.
  
  
  CONCLUSIONS
  These findings confirm that ploidy and SPF are important new prognostic factors that are able to identify selected groups of patients at high risk of treatment failure, even if the tumor's presentation is favorable according to standard criteria.
authors.affiliation: Department of Pediatrics, University of Padua, Italy.
authors.forename: L
authors.initials: L
authors.lastname: De Zen
authors.forename: A
authors.initials: A
authors.lastname: Sommaggio
authors.forename: E S
authors.initials: ES
authors.lastname: d'Amore
authors.forename: L
authors.initials: L
authors.lastname: Masiero
authors.forename: L C
authors.initials: LC
authors.lastname: di Montezemolo
authors.forename: A
authors.initials: A
authors.lastname: Linari
authors.forename: E
authors.initials: E
authors.lastname: Madon
authors.forename: C
authors.initials: C
authors.lastname: Dominici
authors.forename: S
authors.initials: S
authors.lastname: Bosco
authors.forename: G
authors.initials: G
authors.lastname: Bisogno
authors.forename: M
authors.initials: M
authors.lastname: Carli
authors.forename: V
authors.initials: V
authors.lastname: Ninfo
authors.forename: G
authors.initials: G
authors.lastname: Basso
mesh_terms: D000293:Adolescent; D002455:Cell Division; D002648:Child; D002675:Child, Preschool; D004273:DNA, Neoplasm / Q000032:analysis; D005260:Female; D005434:Flow Cytometry; D005557:Formaldehyde; D006801:Humans; D007223:Infant; D008297:Male; D015999:Multivariate Analysis; D016612:Paraffin Embedding; D011003:Ploidies; D012189:Retrospective Studies; D012208:Rhabdomyosarcoma / Q000235:genetics* / Q000473:pathology; D018232:Rhabdomyosarcoma, Alveolar / Q000235:genetics / Q000473:pathology; D018233:Rhabdomyosarcoma, Embryonal / Q000235:genetics / Q000473:pathology; D012680:Sensitivity and Specificity
chemical_list: D004273:DNA, Neoplasm; D005557:Formaldehyde