p27Kip1 induces drug resistance by preventing apoptosis upstream of cytochrome c release and procaspase-3 activation in leukemic cells.
The cyclin-dependent kinase inhibitor p27Kip1 has been implicated as a drug resistance factor in tumor cells grown as spheroids or confluent monolayers. Here, we show that p27Kip1 overexpression also induces resistance to drug-induced apoptosis and cytotoxicity in human leukemic cells growing in suspension. The anti-apoptotic effect of p27Kip1 is not restricted to DNA-damaging agents but extends to the tubulin poison vinblastin, agonistic anti-Fas antibodies and macromolecule synthesis inhibitors. To further identify at which level this protein interferes with the cell death pathway, we investigated its influence on caspase activation and mitochondrial changes. Exposure of mock-transfected U937 cells to 50 microm etoposide activates procaspase-3 and the long isoform of procaspase-2 and induces mitochondrial potential decrease and cytochrome c release from mitochondria to the cytosol. All these events are prevented by p27Kip1 overexpression. p27Kip1 does not modulate Bcl-2, Bcl-X(L), Mcl-1 and Bax protein level in leukemic cells but suppresses Mcl-1 expression decrease observed in mock-transfected U937 cells undergoing etoposide-induced cell death. We conclude that p27Kip1 prevents cell death upstream of the final pathway common to many apoptotic stimuli that involves cytochrome c release from mitochondria and activation of downstream caspases.
Laboratory of Onco-Hematology and Pharmacology, CJF INSERM 94-08, Faculty of Medicine & Pharmacy, Dijon, France.
B
B
Eymin
M
M
Haugg
N
N
Droin
O
O
Sordet
M T
MT
Dimanche-Boitrel
E
E
Solary
D000818:Animals; D017209:Apoptosis; D053143:Caspase 2; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism*; D018797:Cell Cycle Proteins; D050760:Cyclin-Dependent Kinase Inhibitor p27; D003574:Cytochrome c Group / Q000378:metabolism*; D004351:Drug Resistance; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000378:metabolism*; D005047:Etoposide / Q000494:pharmacology; D015870:Gene Expression; D006801:Humans; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis* / Q000235:genetics; D008928:Mitochondria / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D025521:Tumor Suppressor Proteins; D020298:U937 Cells
C493961:Cdkn1b protein, mouse; D018797:Cell Cycle Proteins; D003574:Cytochrome c Group; D004792:Enzyme Precursors; D008869:Microtubule-Associated Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D025521:Tumor Suppressor Proteins; D050760:Cyclin-Dependent Kinase Inhibitor p27; D005047:Etoposide; C505407:CASP3 protein, human; C505409:Casp3 protein, mouse; D053143:Caspase 2; D053148:Caspase 3; D020169:Caspases