Cell cycle regulatory protein expression in fresh acute myeloid leukemia cells and after drug exposure.
Characteristics of treatment-induced cell cycle arrest are important for in vitro and in vivo sensitivity of acute myeloid leukemia (AML) cells to cytotoxic drugs. We analyzed the expression of the major G1 cell cycle regulators (p21Cip1, p27Kip1, cyclins D, cyclin E and pRb) in 41 fresh AML cell samples. The level of p27 expression was the only factor correlated with the response to chemotherapy, a high level of p27 expression being predictive of complete remission. There was a close relation between expression of pRb, cyclin D2 and FAB subtype, illustrated by the absence of both proteins in most samples having a monocytic component (M4, M5). We also assessed the expressions of pRb, cyclin E, p21 and p27 and the activity of cdk2, the major regulator of S-phase entry, after exposure to cytosine-arabinoside (AraC) and daunorubicin (DNR), and found these proteins could characterize time- and dose-dependent cellular response to each drug. We observed hyperphosphorylated pRb, increased levels of cyclin E and a high cdk2 activity, but no p21 induction, in AML cells exposed to 10(-6) M AraC. After exposure to 10(-5) M AraC, corresponding to the serum concentration reached in high-dose AraC regimens (HDAraC), a strong p21 induction was observed, associated with similarly overexpressed cyclin E and even higher cdk2 activity than after 10(-6) M AraC, while apoptosis was significantly increased. These data suggest that cdk2 activity is likely to play a role in AraC-induced apoptosis in AML cells. This mechanism may account for high efficacy of HDAraC in cells showing little sensitivity to conventional AraC doses.
Radosevic
N
N
Hematology Department, INSERM E9912/EA1529, Hotel-Dieu, Paris, France.
Delmer
A
A
Tang
R
R
Marie
J P
JP
Ajchenbaum-Cymbalista
F
F
D017209:Apoptosis; D042846:CDC2-CDC28 Kinases; D018797:Cell Cycle Proteins; D056741:Cyclin D; D019927:Cyclin E / Q000032:analysis*; D051357:Cyclin-Dependent Kinase 2; D050759:Cyclin-Dependent Kinase Inhibitor p21; D050760:Cyclin-Dependent Kinase Inhibitor p27; D018844:Cyclin-Dependent Kinases / Q000032:analysis; D016213:Cyclins / Q000032:analysis*; D003561:Cytarabine / Q000494:pharmacology; D004249:DNA Damage; D003630:Daunorubicin / Q000494:pharmacology; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology; D015470:Leukemia, Myeloid, Acute / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D008869:Microtubule-Associated Proteins / Q000032:analysis*; D017346:Protein Serine-Threonine Kinases / Q000032:analysis; D016160:Retinoblastoma Protein / Q000032:analysis*; D019089:Stem Cell Factor / Q000494:pharmacology; D025521:Tumor Suppressor Proteins
C493956:CDKN1A protein, human; D018797:Cell Cycle Proteins; D056741:Cyclin D; D019927:Cyclin E; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D007377:Interleukin-3; D008869:Microtubule-Associated Proteins; D016160:Retinoblastoma Protein; D019089:Stem Cell Factor; D025521:Tumor Suppressor Proteins; D003561:Cytarabine; D050760:Cyclin-Dependent Kinase Inhibitor p27; D017346:Protein Serine-Threonine Kinases; D042846:CDC2-CDC28 Kinases; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases; D003630:Daunorubicin