Cell cycle-related expression of p120 nucleolar antigen in normal human lymphocytes and in cells of HL-60 and MOLT-4 leukemic lines: effects of methotrexate, camptothecin, and teniposide.
Expression of the proliferation-associated nucleolar antigen p120 was studied by flow cytometry in human quiescent and phytohemagglutinin-stimulated lymphocytes, as well as in human lymphocytic (MOLT-4) and promyelocytic (HL-60) cell lines. Bivariate analysis of p120 and DNA content made it possible to correlate p120 expression with cell position in the cycle. Proliferating lymphocytes and MOLT-4 and HL-60 cells had a similar pattern of p120 expression. Populations of G1 cells, in all three cell types, were very heterogenous with respect to p120, and a threshold in G1 was observed. The cells with a p120 level below the threshold value did not enter S phase. An increase in p120 was observed during progression through S phase, and the antigen was maximally expressed in G2 cells. The p120/DNA content ratio, however, was highest in late G1 cells (G1B) and was declining during S and G2. The data thus suggest that p120 may be degraded during mitosis and that the postmitotic cells inherit little, if any, of this protein; the antigen then accumulates predominantly during G1, and must reach a threshold level to enable the cells to enter S phase. Antigen p120 could not be detected in noncycling lymphocytes nor in HL-60 cells induced to myeloid differentiation by growth in the presence of dimethyl sulfoxide. Treatment of MOLT-4 cells with pharmacological concentrations of methotrexate, camptothecin, or teniposide induced cell arrest in S or G2; expression of p120 in the arrested cells was unchanged from that of untreated MOLT-4 controls at the same phase of the cycle. The level of p120 was minimal in MOLT-4 or HL-60 cells arrested in M phase by vinblastine, but vinblastine had no effect on p120 fluorescence of interphase cells. Camptothecin or teniposide induced apoptosis selectively in S phase of HL-60 cells; apoptotic cells from camptothecin-treated cultures, however, despite the marked nucleolysis, still expressed p120. The data on the drug-treated cells indicate that the p120 level in tumors of patients may be used as a marker of tumor/malignancy even in clinical samples obtained during treatment.
Gorczyca
W
W
Cancer Research Institute, New York Medical College, Valhalla 10595.
Bruno
S
S
Melamed
M R
MR
Darzynkiewicz
Z
Z
D000951:Antigens, Neoplasm / Q000378:metabolism*; D002166:Camptothecin / Q000494:pharmacology*; D002453:Cell Cycle; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000276:immunology* / Q000473:pathology; D008214:Lymphocytes / Q000166:cytology / Q000276:immunology*; D008727:Methotrexate / Q000494:pharmacology*; D009687:Nuclear Proteins / Q000378:metabolism*; D013713:Teniposide / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D014747:Vinblastine / Q000494:pharmacology*; D012359:tRNA Methyltransferases
D000951:Antigens, Neoplasm; D009687:Nuclear Proteins; D014747:Vinblastine; D013713:Teniposide; C065028:NOP2 protein, human; D012359:tRNA Methyltransferases; D002166:Camptothecin; D008727:Methotrexate