Induction of p53 and apoptosis by delta 12-PGJ2 in human hepatocarcinoma SK-HEP-1 cells.
Human hepatocarcinoma cells (SK-HEP-1) were induced to die through apoptosis by treatment with delta 12-prostaglandin (PG)J2, as characterized by the appearance of a typical DNA ladder. The induction of apoptosis by delta 12-PGJ2 was specifically blocked by cycloheximide (CHX). Western analysis using anti-p53 or anti-WAF1 monoclonal antibodies demonstrated that these two protein levels were increased 3 h after delta 12-PGJ2 treatment, and accumulated for up to 12 h. The induction of p53 protein seemed to be dependent on the increase of p53 mRNA level, which was inhibited by CHX treatment. However, delayed addition of CHX after delta 12-PGJ2 treatment failed to affect both p53 mRNA levels and DNA fragmentation following delta 12-PGJ2 treatment, indicating that the inhibition of p53 synthesis may contribute to the protective effect of CHX against delta 12-PGJ2-mediated cytotoxicity. Therefore, our results suggest that the initial events caused by delta 12-PGJ2, leading ultimately to SK-HEP-1 cell death, involve a certain process required for p53 induction. However, the finding that delta 12-PGJ2 is also active against Hep 3B cells which are devoid of a functional p53 indicates that p53 may not be the critical requirement for inducing apoptosis by delta 12-PGJ2.
Lee
J H
JH
Department of Biochemistry, Catholic University Medical College, Seoul, South Korea.
Kim
H S
HS
Jeong
S Y
SY
Kim
I K
IK
D017209:Apoptosis / Q000187:drug effects*; D006528:Carcinoma, Hepatocellular; D002470:Cell Survival; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000096:biosynthesis; D003513:Cycloheximide / Q000494:pharmacology; D004273:DNA, Neoplasm / Q000378:metabolism; D006801:Humans; D015230:Prostaglandin D2 / Q000031:analogs & derivatives* / Q000494:pharmacology; D011465:Prostaglandins, Synthetic / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis*
C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D004273:DNA, Neoplasm; D011465:Prostaglandins, Synthetic; D012333:RNA, Messenger; D016159:Tumor Suppressor Protein p53; C037112:9-deoxy-delta-9-prostaglandin D2; D003513:Cycloheximide; D015230:Prostaglandin D2