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6th ESACP Congress, Heidelberg, April 7-11, 1999 |
A096
Two ways are possible to measure cell proliferation. The first one consists
of quantifying the number of cycling cells with the help of antibodies
directed against cells either in G1, S, G2 or M phase. The second way is
to assess the cell cycle duration by the quantification of AgNOR proteins.
Measuring both features on the same slide represents an attractive way to
tackle with the proliferating activity of a cell culture or a tumor. Here,
we propose a MIB-1 and AgNOR double staining method especially adapted to
image cytometry measurement, using MIB-1 antibody coupled to FITC in order
to avoid the thresholding problems encountered with such a multilabeling
technique. We have applied this new method on a series of 39 breast cancer
cases with at least 4 years follow-up, in order to determine the prognosis
significance of this measurement. MIB-1 alone is not linked to prognosis,
while the global mean AgNOR area is significantly linked to prognosis in
terms of development of visceral metastasis or death. However, the global
mean AgNOR area is insufficient to determine the time limit of appearance
of metastasis or relapse. We clearly demonstrate that a high mean AgNOR area
within a cell population having a high MIB-1 index can discriminate tumors
with a high metastatic potential. By multiplying AgNOR area by the percentage
of MIB-1 positive cells we calculate the proliferative activity, (P), which
brings very important informations concerning the time limit of relapse.
A DOUBLE-STAINING TECHNIQUE FOR AGNOR QUANTIFICATION IN MIB-1 POSITIVE
CELLS ESPECIALLY ADAPTED FOR IMAGE CYTOMETRY
Lorenzato M, Abboud P, Lechki C, Browarnij F, O'Donohue MF*,
Adnet JJ, Ploton D*
Lab. Pol Bouin, CHU Maison Blanche, and Unité MEDIAN, EA 2063*, IFR 53,
Reims, France