Protease inhibitor-induced apoptosis: accumulation of wt p53, p21WAF1/CIP1, and induction of apoptosis are independent markers of proteasome inhibition.
Inhibitors of proteases are currently emerging as a potential anti-cancer modality. Nonselective protease inhibitors are cytotoxic to leukemia and cancer cell lines and we found that this cytotoxicity is correlated with their potency as inhibitors of the proteasome but not as inhibitors of calpain and cathepsin. Highly selective inhibitors of the proteasome were more cytotoxic and fast-acting than less selective inhibitors (PS341>>ALLN>>ALLM). Induction of wt p53 correlated with inhibition of the proteasome and antiproliferative effect in MCF7, a breast cancer cell line, which was resistant to apoptosis caused by proteasome inhibitors. In contrast, inhibitors of the proteasome induced apoptosis in four leukemia cell lines lacking wt p53. The order of sensitivity of leukemia cells was: Jurkat>HL60> or =U937>>K562. The highly selective proteasome inhibitor PS-341 induced cell death with an IC50 as low as 5 nM in apoptosis-prone leukemia cells. Cell death was preceded by p21WAF1/CIP1 accumulation, an alternative marker of proteasome inhibition, and by cleavage of PARP and Rb proteins and nuclear fragmentation. Inhibition of caspases abrogated PARP cleavage and nuclear fragmentation and delayed, but did not completely prevent cell death caused by PS-341. Reintroduction of wt p53 into p53-null PC3 prostate carcinoma cells did not increase their sensitivity to proteasome inhibitors. Likewise, comparison of parental and p21-deficient cells demonstrated that p21WAF1/CIP1 was dispensable for proteasome inhibitor-induced cytotoxicity. We conclude that accumulation of wt p53 and induction of apoptosis are independent markers of proteasome inhibition.
An
W G
WG
Medicine Branch, National Cancer Institute, NIH, Bethesda, MD 20892, USA.
Hwang
S G
SG
Trepel
J B
JB
Blagosklonny
M V
MV
D000111:Acetylcysteine / Q000031:analogs & derivatives / Q000494:pharmacology / Q000633:toxicity; D000179:Acrylates / Q000494:pharmacology / Q000633:toxicity; D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology / Q000633:toxicity; D017209:Apoptosis / Q000187:drug effects*; D001897:Boronic Acids / Q000494:pharmacology / Q000633:toxicity; D000069286:Bortezomib; D002154:Calpain / Q000037:antagonists & inhibitors; D002403:Cathepsins / Q000037:antagonists & inhibitors; D002455:Cell Division / Q000187:drug effects; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000235:genetics / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D004357:Drug Synergism; D016158:Genes, p53; D006801:Humans; D019169:Jurkat Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D007976:Leupeptins / Q000494:pharmacology / Q000633:toxicity; D009097:Multienzyme Complexes / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D009369:Neoplasms / Q000473:pathology; D009842:Oligopeptides / Q000494:pharmacology / Q000633:toxicity; D011480:Protease Inhibitors / Q000494:pharmacology* / Q000633:toxicity; D046988:Proteasome Endopeptidase Complex; D011719:Pyrazines / Q000494:pharmacology / Q000633:toxicity; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*; D020298:U937 Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism
D000179:Acrylates; D000590:Amino Acid Chloromethyl Ketones; D001897:Boronic Acids; C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D007976:Leupeptins; D009097:Multienzyme Complexes; D009363:Neoplasm Proteins; D009842:Oligopeptides; C100442:PD 150606; D011480:Protease Inhibitors; D011719:Pyrazines; D016159:Tumor Suppressor Protein p53; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; C063261:acetylleucyl-leucyl-norleucinal; C069861:calpain inhibitor 2; C067713:lactacystin; D000069286:Bortezomib; D002403:Cathepsins; D002154:Calpain; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; D000111:Acetylcysteine