Absence of radiation-induced G1 arrest in two closely related human lymphoblast cell lines that differ in p53 status.
In order to examine more precisely the role of p53 in the activation of the G1/S checkpoint by ionizing radiation, we examined two human lymphoblast cell lines derived from the same donor. The TK6 line had a doubling time of 12.2 h and expressed wild type p53, while the WTK1 line had a doubling time of 12.7 h and expressed mutant p53. The two lines differ significantly in their susceptibility to radiation-induced cell killing and apoptosis. Cells were examined by flow cytometry at regular intervals from 0 to 12 h after irradiation with two different doses designed to yield equivalent survival levels in both cell lines. In some experiments, cells were incubated with colcemid to block them in the first postirradiation mitosis and prevent contamination of the flow cytometric profiles with second cycle cells. There was no significant difference between the two cell lines in the progression of irradiated cells out of G1 and into the S and G2 phases of the cell cycle. In particular, there was no evidence for a prolonged arrest in G1 in the TK6 cell line expressing wild type p53. Furthermore, expression of the p53 downstream genes WAF1/CIP1 and RB appeared normal in TK6 cells. These results suggest that factors other than those in the p53 signal transduction pathway alone may be required to activate the G1/S checkpoint in irradiated human cells and that apoptosis and G1 arrest may utilize different pathways.
Little
J B
JB
Department of Cancer Biology, Harvard School of Public Health, Boston, Massachusetts 02115, USA.
Nagasawa
H
H
Keng
P C
PC
Yu
Y
Y
Li
C Y
CY
D017209:Apoptosis / Q000528:radiation effects*; D001483:Base Sequence; D002453:Cell Cycle / Q000502:physiology / Q000528:radiation effects*; D002460:Cell Line; D002470:Cell Survival / Q000502:physiology / Q000528:radiation effects; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000096:biosynthesis / Q000235:genetics; D017931:DNA Primers; D004307:Dose-Response Relationship, Radiation; D005091:Exons; D005434:Flow Cytometry; D016193:G1 Phase / Q000528:radiation effects; D016161:Genes, Retinoblastoma; D016158:Genes, p53; D006801:Humans; D008214:Lymphocytes; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D047428:Protein Kinase Inhibitors; D015398:Signal Transduction
C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D017931:DNA Primers; D047428:Protein Kinase Inhibitors