Aim of the study: Evaluation of significance of investigation of Ki67 and AgNOR staining as an indices of proliferative activity of megakaryocytes.

Material and methods: Bone marrow smears from 151 patients suffering from hematological malignancies: polycythemia vera (PV) − 14 cases, myelofibrosis (MF) − 14, essential thrombocythemia (ET) − 15, chronic myeloid leukemia (CML) − 11, myelodysplastic syndromes (MDS) − 18; acute myeloid leukemia (AML) − 30, acute lymphoblastic leukemia (ALL) − 10, low grade non-Hodgkin’s lymphomas (NHL) with bone marrow infiltration − 20, multiple myeloma (MM) − 19. Cytological slides were stored in the room temperature more than 24 hours and then stained using immunocytochemical method for Ki67 and cytochemical method for AgNOR. Slides were analyzed using illuminating microscope with digital image analysis (software LUCIA). Mean particles area, mean nuclear area and maturity of nucleus in AgNOR staining as well as intensity of Ki67 expression were evaluated, according to arbitrary scale (strong, medium, light, no reaction).

Results: Look at the table 1 and table 2.

Conclusions:There is a good correlation between expression of Ki-67 and AgNOR’s in megakaryocytes as well as between nuclear maturity and indices of Ki-67 and AgNOR in myeloproliferative disorders. Differences in AgNOR’s and Ki 67 staining area between myeloproliferative diseases and another hematological malignancies are also significant.

Table 1:

Values of indices of the Ki67 and AgNOR in megakaryocytes

DiseasesMean nucleus surface μm2Mean AgNOR particle area in nucleus of megakaryocytes μm2Mean Ki67 particle area in nucleus of megakaryocytes μm2Intensity of Ki67 expression in megakaryocytesGrade of megakaryocytes maturity
PV 667,29 0,68 0,19 Medium III and IV 
ET 812,32 0,82 0,29 Strong / Medium III and IV 
MF 767,92 0,55 0,26 Strong / Medium III / IV 
CML 734,30 0,57 0,15 Medium / Light III / IV 
MDS 645,67 0,06 0,03 Light III / IV 
AML 679,04 0,04 No reaction No reaction IV 
ALL 712,23 0,03 No reaction No reaction IV 
NHL 798,56 0,04 No reaction No reaction IV 
MM 629,49 0,04 No reaction No reaction IV 
DiseasesMean nucleus surface μm2Mean AgNOR particle area in nucleus of megakaryocytes μm2Mean Ki67 particle area in nucleus of megakaryocytes μm2Intensity of Ki67 expression in megakaryocytesGrade of megakaryocytes maturity
PV 667,29 0,68 0,19 Medium III and IV 
ET 812,32 0,82 0,29 Strong / Medium III and IV 
MF 767,92 0,55 0,26 Strong / Medium III / IV 
CML 734,30 0,57 0,15 Medium / Light III / IV 
MDS 645,67 0,06 0,03 Light III / IV 
AML 679,04 0,04 No reaction No reaction IV 
ALL 712,23 0,03 No reaction No reaction IV 
NHL 798,56 0,04 No reaction No reaction IV 
MM 629,49 0,04 No reaction No reaction IV 
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Table 2:

Intensity of Ki67 expression in megakaryocytes in hematological disorders

DiseasesStrong – % of cellsMedium – % of cellsLight – % of cellsNo reaction – % of cells
PV 36 43 14 7 
ET 56 33 11 0 
MF 51 32 16 1 
CML 16 43 34 7 
MDS 9 12 68 11 
AML 0 0 0 0 
ALL 0 0 0 0 
NHL 0 0 0 0 
MM 0 0 0 0 
DiseasesStrong – % of cellsMedium – % of cellsLight – % of cellsNo reaction – % of cells
PV 36 43 14 7 
ET 56 33 11 0 
MF 51 32 16 1 
CML 16 43 34 7 
MDS 9 12 68 11 
AML 0 0 0 0 
ALL 0 0 0 0 
NHL 0 0 0 0 
MM 0 0 0 0 
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Topics:
acute lymphocytic leukemia, digital image analysis, hematologic neoplasms, hematological diseases, hodgkin's disease, ki-67 antigen, leukemia, myelocytic, acute, leukemia, myelocytic, chronic, lymphoma, lymphoproliferative disorders

Author notes

Corresponding author

2005, The American Society of Hematology
2005
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