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 surface phenotypes and cell cycle of 22 hematopoietic cell lines based on their cellular DNA/RNA content. Populations of G1a, G1b, S, and G2M, the DNA index (DI), and the RNA index of S phase (SRI) were calculated 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) and the coefficient of variations in the RNA at S phase (SVC). DNA/RNA dot plots of cell lines revealed four characteristic profiles of the cell 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 O included four stem cell lines: one t(1;19) leukemia, two Ph1+ acute lymphocytic leukemia (ALL), and one biphenotypic crisis of chronic granulocytic leukemia (CGL). Type I included five ALL cell lines: three T-ALL and two common B-ALL. Type II contained 10 myeloid cell lines: five AML and five 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-cycle analysis of blasts using 7AAD/PY combined with surface phenotyping may yield important information for classifying hematopoietic malignancy within 2 hours of patient admission.
Toba
K
K
The First Department of Internal Medicine, Niigata University School of Medicine, Japan.
Kishi
K
K
Koike
T
T
Winton
E F
EF
Takahashi
H
H
Nagai
K
K
Maruyama
S
S
Furukawa
T
T
Hashimoto
S
S
Masuko
M
M
Uesugi
Y
Y
Kuroha
T
T
Tsukada
N
N
Shibata
A
A
D015703:Antigens, CD / Q000032:analysis; D002453:Cell Cycle; D002460:Cell Line; D004273:DNA, Neoplasm / Q000032:analysis*; D003609:Dactinomycin / Q000031:analogs & derivatives; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D016130:Immunophenotyping; D007938:Leukemia / Q000276:immunology / Q000473:pathology*; D008223:Lymphoma / Q000276:immunology / Q000473:pathology*; D011754:Pyronine; D012334:RNA, Neoplasm / Q000032:analysis*; D013194:Staining and Labeling; D014407:Tumor Cells, Cultured
D015703:Antigens, CD; D004273:DNA, Neoplasm; D012334:RNA, Neoplasm; D003609:Dactinomycin; C025942:7-aminoactinomycin D; D011754:Pyronine