PROFILE OF CELL-CYCLE IN HEMATOPOIETIC MALIGNANCY BY DNA RNA QUANTITATION USING 7AAD/PY/

Using 7-amino-actinomycin-D/pyronin Y (7AAD/PY), we analyzed the surfa ce phenotypes and cell cycle of 22 hematopoietic cell lines based on t heir cellular DNA/RNA content. Populations of G(1a), G(1b), S, and G(2 )M, the DNa index (DI), and the RNA index of S phase (SRI) were calcul ated by means of DNA/RNA dot plots. Two new parameters were extracted from the cell-cycle profiles: the nucleic acid index of S phase (NI) a nd the coefficient of variations in the RNA at S phase (SCV). DNA/RNA dot plots of cell lines revealed four characteristic profiles of the c ell cycle, defined with the calculated NI and SCV. These were type 0 ( small NI, large SCV), type I (small NI, small SCV), type II (large NI, small SCV), and type III (large NI, large SCV). Type 0 included four stem cell lines: one t(1;19) leukemia, two Ph(1+) acute lymphocytic le ukemia (ALL), and one biphenotypic crisis of chronic granulocytic leuk emia (CGL). Type I included five ALL cell lines: three T-ALL and two c ommon B-ALL. Type II contained 10 myeloid cell lines: five AML and fiv e myeloid crisis of CGL. Type III contained three relatively immature lymphoma cell lines: two Burkitt's lymphoma and one follicular center lymphoma. Calculated NI/SCV (%) were as follows: type 0, 2.27 +/- 0.19 /16.7 +/- 3.7; type I, 2.20 +/- 0.30/11.1 +/- 0.7; type II, 3.64 +/- 0 .52/11.8 +/- 1.0; and type III, 3.60 +/- 0.53/17.5 +/- 1.9. Cell-analy sis of blasts using 7AAD/PY combined with surface phenotyping may yiel d important information for classifying hematopoietic malignancy withi n 2 hours of patient admission.