Abstract
The proliferation markers, S-phase fraction (SPF) and MIB1, were evaluated in 90 primary node-negative breast carcinomas. SPF was determined flow cytometrically using an improved Hedley protocol for release of pure nuclei from formalin-fixed, paraffin-embedded sections. SPF analysis was performed using the ModFit software program (Verity, Maine, U.S.A.). MIB1 immunostaining (Dianova, Hamburg, Germany) was performed on adjacent paraffin sections using APAAP. The percentage of MIB1 positive cells was calculated in 500 randomly chosen tumor cells. Median follow-up was 34 (9–72) months. An optimal cutoff of 8% SPF was found using isotonic regression analysis: 61 (68%) tumors had low (≤ 8%) and 29 (32%) high (> 8%) SPF. The optimal cutoff for MIB1 was 25%: 75 (83%) tumors had a low (≤ 25%) and 15 (17%) a high (> 25%) MIB1 staining percentage. MIB1 was significantly correlated to grading (P = 0.018) and medullary histological tumor type (P
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