title: Prognostic value of DNA cytometry in 281 premenopausal patients with lymph node negative breast carcinoma randomized in a control trial: multivariate analysis with Ki-67 index, mitotic count, and microvessel density.
abstract: BACKGROUND
  The clinical relevance of DNA image cytometry (ICM) and flow cytometry (FCM) remains under investigation in breast carcinoma. The objective of the current work was to study the prognostic value of DNA ICM and FCM in a series of patients randomized in a control trial. A multivariate analysis has been performed including other factors still under investigation such as Ki-67 index, mitotic count, microvessel density, and P53 and Bcl-2 expression.
  
  
  METHODS
  Two hundred and eighty-one patients were randomized in the European Organization for Research and Treatment of Cancer 10854 trial comparing surgery followed by one course of perioperative chemotherapy versus surgery alone. Tumor parameters studied were pT, multicentricity, tumor grading according to modified Scarff-Bloom-Richardson, estrogen receptors, mitotic count per 1.7 mm(2), MIB-1, and BCL-2 scores, microvessel density, and p53 expression. ICM DNA parameters studied from paraffin embedded specimens, were DNA ploidy, proliferative index, 2c deviation index, malignancy grade, and Auer-Baldetorp typing. FCM DNA parameters analyzed on the same samples were ploidy and S-phase fraction statistics. The influence of tumor parameters, and DNA parameters on overall survival (OS), disease free survival (DFS), and metastasis-free survival (MFS) was evaluated using the Cox model. Median follow-up was 82 months.
  
  
  RESULTS
  For OS, the prognostic parameters retained were pathologic tumor size (pT) and mitotic index (MI). Overall survival was 94% and 68% for tumors pT1/MI less than 10 and pT2-3 MI greater than or equal to 10, respectively. For DFS, age, multicentricity, and grading according to modified Scarff and Bloom were predicting factors with the same relative risk. Disease free survival was 96%, 78% and 68% respectively, when 1, 2, or 3 of those factors were present. For MFS, the only retained predicting factor was MI. MFS was 97% and 73% when MI was less than 10 and MI was greater than or equal to 10, respectively.
  
  
  CONCLUSIONS
  Evaluation of proliferative compartment was the most important predicting factor for OS and MFS in the current series of premenopausal lymph node negative patients with breast invasive carcinoma. When working on paraffin embedded tissue, the best way of assessing it was MI count. ICM DNA analysis results were not selected in multivariate analysis. DNA analysis by FCM should be considered as an unsuitable technique when working on paraffin embedded tissue.
authors.affiliation: Department of Pathology, Centre François Baclesse, Caen, France. am.mandard@baclesse.fr
authors.forename: A M
authors.initials: AM
authors.lastname: Mandard
authors.forename: Y
authors.initials: Y
authors.lastname: Denoux
authors.forename: P
authors.initials: P
authors.lastname: Herlin
authors.forename: F
authors.initials: F
authors.lastname: Duigou
authors.forename: M J
authors.initials: MJ
authors.lastname: van De Vijver
authors.forename: P C
authors.initials: PC
authors.lastname: Clahsen
authors.forename: L
authors.initials: L
authors.lastname: van Den Broek
authors.forename: T M
authors.initials: TM
authors.lastname: Sahmoud
authors.forename: M
authors.initials: M
authors.lastname: Henry-Amar
authors.forename: C J
authors.initials: CJ
authors.lastname: van De Velde
mesh_terms: D000782:Aneuploidy; D001943:Breast Neoplasms / Q000188:drug therapy / Q000473:pathology* / Q000601:surgery*; D003131:Combined Modality Therapy; D004273:DNA, Neoplasm / Q000032:analysis*; D004171:Diploidy; D018572:Disease-Free Survival; D005260:Female; D006801:Humans; D019394:Ki-67 Antigen / Q000032:analysis*; D008833:Microcirculation / Q000473:pathology; D008875:Middle Aged; D008940:Mitotic Index; D015999:Multivariate Analysis; D009361:Neoplasm Invasiveness; D009367:Neoplasm Staging; D011003:Ploidies; D017697:Premenopause; D011960:Receptors, Estrogen / Q000032:analysis; D016159:Tumor Suppressor Protein p53 / Q000032:analysis
chemical_list: D004273:DNA, Neoplasm; D019394:Ki-67 Antigen; D011960:Receptors, Estrogen; D016159:Tumor Suppressor Protein p53