Proliferating cell nuclear antigen (PCNA)/cyclin expression during the cell cycle in normal and leukemic cells.
Bivariate flow cytometric analysis of the cell proliferation-associated nuclear protein, identified as the "proliferating cell nuclear antigen" (PCNA)/cyclin and of nuclear DNA content, was performed in quiescent and mitogen-stimulated human peripheral blood lymphocytes, in EUE (human embryonic epithelium) cells, before and after a long-term exposure to a hypertonic (HT) medium, in 4 human leukemic cell lines and in fresh bone marrow (BM) cells from 10 patients with untreated acute non-lymphoblastic leukemia (ANLL). The PCNA/cyclin was detected using both an autoantibody extracted from sera of systemic lupus erythematosus patients and the recently produced mouse monoclonal antibody (MoAb) IgG, named 19F4. The distribution of cells in the different phases of the cycle and the percentage of S-phase cells were obtained in duplicate samples, by DNA flow cytometry (FCM) and by dual parameter FCM of DNA content and bromodeoxyuridine (BUDR) incorporation. In all cell types, the non-specific cytoplasmic background fluorescence was significantly lower with the MoAb compared to that obtained with the polyclonal Ab. The percentage of PCNA-positive cells (both with the autoantibody and the 19F4 MoAb) was always higher than that of S-phase cells by DNA FCM and of BUDR-labeled cells. The pattern of PCNA-expression in both normal proliferating cells and acute leukemia cells, showed that most G0/G1 cells did not express significant amounts of PCNA; an increase in PCNA immunofluorescence was found in late G1 cells, and further increases were observed in S- and G2-M phase cells. PCNA/cyclin, as revealed both with autoantibodies and with the 19F4 MoAb, is associated with all actively or potentially dividing (i.e. G1, S and G2-M) cells thus identifying the proliferative cellular compartment. Combined with the use of multiparameter FCM techniques, the PCNA immunolocalization offers a useful tool to study cell kinetics in normal and leukemic human cell populations.
Giordano
M
M
Department of Internal Medicine, University and I.R.C.C.S. San Matteo, Pavia, Italy.
Danova
M
M
Pellicciari
C
C
Wilson
G D
GD
Mazzini
G
G
Conti
A M
AM
Franchini
G
G
Riccardi
A
A
Romanini
M G
MG
D000911:Antibodies, Monoclonal / Q000276:immunology; D001854:Bone Marrow Cells; D001973:Bromodeoxyuridine / Q000378:metabolism; D002453:Cell Cycle; D002460:Cell Line; D004247:DNA / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods; D005455:Fluorescent Antibody Technique; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000378:metabolism* / Q000473:pathology; D008214:Lymphocytes / Q000187:drug effects / Q000378:metabolism; D009687:Nuclear Proteins / Q000276:immunology / Q000378:metabolism*; D018809:Proliferating Cell Nuclear Antigen
D000911:Antibodies, Monoclonal; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen; D004247:DNA; D001973:Bromodeoxyuridine