Contributions of p53 and PMA to gamma-irradiation induced apoptosis in Jurkat cells.
Several mutations prevent the expression of p53 in the human lymphoblastoid T cell line Jurkat. Restoration of p53 in Jurkat cells had no effect on the cell growth but markedly increased the amount of apoptosis induced by gamma-irradiation. Inhibition of RNA synthesis using 5,6-dichlorobenimidizole riboside had little effect on apoptosis induced by irradiation in the presence of p53 and did not affect the p53-independent apoptotic pathway. Expression of p53 also had no effect on the expression levels of proteins such as Fas, GADD45, Bax, Bcl-2, Bcl-x(L) or p53 induced proteins (PIGS) in resting cells or after irradiation. Activation of protein kinase C by phorbol 12-myristate 13-acetate produced an almost complete inhibition of p53-independent apoptosis following irradiation, whereas no significant effect was observed on the rate of p53-induced apoptosis. Although phorbol 12-myristate 13-acetate strongly induced p21 and stabilised p53 in the resting transfected Jurkat cells, neither apoptosis nor cell arrest was observed. In summary, this work shows that p53 enhances the radiosensitivity of Jurkat cells through an apoptotic process that is triggered by irradiation and is largely independent of RNA synthesis and protein kinase C activation. Apoptosis in p53- negative Jurkat cells is strongly inhibited by PMA indicating that the pathway triggered by p53 may be distinct from apoptotic pathways used in its absence.
Vigorito
E
E
Servei d'Immunologia, Hospital Clinic, Barcelona, Spain.
Plaza
S
S
Mir
L
L
Mongay
L
L
Viñas
O
O
Serra-Pagès
C
C
Vives
J
J
D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics / Q000528:radiation effects*; D005720:Gamma Rays; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D019169:Jurkat Cells / Q000187:drug effects* / Q000502:physiology*; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D013997:Time Factors; D014158:Transcription, Genetic; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000187:drug effects / Q000235:genetics / Q000494:pharmacology*
D016159:Tumor Suppressor Protein p53; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate