title: The p27-Skp2 axis mediates glucocorticoid-induced cell cycle arrest in T-lymphoma cells.
abstract: Glucocorticoid therapy is an important treatment modality of hematological malignancies, especially T-cell acute lymphoblastic leukemia (T-ALL). Glucocorticoids are known to induce a cell cycle arrest and apoptosis in T-lymphoma cells. We could demonstrate that the cell cycle arrest induced by the synthetic glucocorticoid dexamethasone (Dex) clearly precedes apoptosis in human CEM T-ALL and murine S49.1 T-lymphoma cells. Cyclin D3 is strongly downregulated, whereas the CDK inhibitor p27 (Kip1) (p27) is strongly upregulated in response to dexamethasone in these cells. RNAi-mediated knockdown of p27 as well as overexpression of its negative regulator Skp2 revealed the critical function of p27 in the Dex-induced G 1 arrest of CEM cells. Our studies indicate that several mechanisms contribute to the increase of p27 protein in our T-lymphoma cell lines. We found a significant upregulation of p27 mRNA in S49.1 and CEM cells. In addition, Dex treatment activated the mouse p27 promotor in reporter gene experiments, indicating a transcriptional regulation. However, the relatively moderate induction of p27 mRNA levels by Dex did not explain the strong increase of p27 protein in CEM and S49.1 cells. We found clear evidence for a posttranslational mechanism responsible for the robust increase in p27 protein. Dex treatment of S49.1 and CEM cells increases the half-life of p27 protein, which indicates that decreased protein degradation is the primary mechanism of p27 induction by glucocorticoids. Interestingly, we found that Dex treatment decreased the protein and mRNA levels of the negative regulator of p27 protein and E3 ubiquitin ligase subunit Skp2. We conclude that the cell cycle inhibitor p27 and its negative regulator Skp2 are key players in the glucocorticoid-induced growth suppression of T-lymphoma cells and should be considered as potential drug targets to improve therapies of T-cell malignancies.
authors.affiliation: Division of Medical Biochemistry, Biocenter, Innsbruck Medical University, Innsbruck, Austria. michael.kullmann@i-med.ac.at
authors.forename: Michael K
authors.initials: MK
authors.lastname: Kullmann
authors.forename: Claudia
authors.initials: C
authors.lastname: Grubbauer
authors.forename: Katrin
authors.initials: K
authors.lastname: Goetsch
authors.forename: Heidelinde
authors.initials: H
authors.lastname: Jäkel
authors.forename: Silvio R
authors.initials: SR
authors.lastname: Podmirseg
authors.forename: Alexander
authors.initials: A
authors.lastname: Trockenbacher
authors.forename: Christian
authors.initials: C
authors.lastname: Ploner
authors.forename: Andrew C B
authors.initials: AC
authors.lastname: Cato
authors.forename: Carsten
authors.initials: C
authors.lastname: Weiss
authors.forename: Reinhard
authors.initials: R
authors.lastname: Kofler
authors.forename: Ludger
authors.initials: L
authors.lastname: Hengst
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D059447:Cell Cycle Checkpoints / Q000187:drug effects*; D045744:Cell Line, Tumor; D056743:Cyclin D3 / Q000378:metabolism; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D003907:Dexamethasone / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects*; D005938:Glucocorticoids / Q000378:metabolism / Q000494:pharmacology*; D006207:Half-Life; D006801:Humans; D015151:Immunoblotting; D008156:Luciferases; D051379:Mice; D054218:Precursor T-Cell Lymphoblastic Leukemia-Lymphoma / Q000188:drug therapy*; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D044786:S-Phase Kinase-Associated Proteins / Q000378:metabolism
chemical_list: D056743:Cyclin D3; D017931:DNA Primers; D005938:Glucocorticoids; D044786:S-Phase Kinase-Associated Proteins; D050760:Cyclin-Dependent Kinase Inhibitor p27; D003907:Dexamethasone; D008156:Luciferases
keywords: Skp2; T-ALL; cell cycle control; dexamethasone; glucocorticoid; lymphoma; p27Kip1; proliferation
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