title: Pharmacological inhibition of cyclin dependent kinases causes p53 dependent apoptosis in renal cell carcinoma.
abstract: OBJECTIVE
  We evaluated the effect of roscovitine (Sigma-Aldrich®), a pharmacological inhibitor of cyclin dependent kinase, on renal cell carcinoma cell lines in vitro.
  
  
  METHODS
  We exposed several renal cell carcinoma cell lines to roscovitine and examined apoptotic signaling pathways using immunoblotting and immunohistochemistry.
  
  
  RESULTS
  As expected, roscovitine caused dose and time dependent inhibition of cyclin dependent kinase 2 autophosphorylation, and of cyclin dependent kinase mediated Pol II phosphorylation in the ACHN (p53-wt) and 786-O (p53 inactive) renal cell carcinoma cell lines (ATCC®). Roscovitine also induced apoptosis in each cell line within a narrow concentration range (about 10 μg/ml). Apoptosis induction was more efficient in ACHN than in 786-O cells and at least partly due to p53 activity. In ACHN cells roscovitine induced apoptosis was associated with p21 induction, and decreased Akt1, XIAP and phospho-Rb expression. These changes also depended on p53 and were not present (p21) or showed a different dose pattern (Akt1, XIAP and phospho-Rb) in 786-O cells. Partial restoration of roscovitine induced apoptosis in 786-O cells by the Mdm-2 inhibitor nutlin-3 (Sigma-Aldrich) suggests that the inactivating mutation of VHL in these cells and its destabilizing effect on p53 are responsible for the decreased sensitivity to apoptosis.
  
  
  CONCLUSIONS
  Our data extend previous studies documenting the pro-apoptotic effect of roscovitine and to our knowledge show for the first time that this activity is restricted to a narrow dose range in renal cell carcinoma cells and partly depends on p53. Thus, roscovitine is a novel potential chemotherapy in a subset of patients with renal cell carcinoma if a narrow therapeutic window is used. These data also provide insight into the role of VHL mutation and p53 in the renal cell carcinoma response to therapeutic cyclin dependent kinase manipulation.
authors.affiliation: Division of Nephrology, Department of Internal Medicine and Cancer Center, University of California-Davis, Davis, California 95616, USA.
authors.forename: Tatsuto
authors.initials: T
authors.lastname: Ishimaru
authors.forename: Jasmine
authors.initials: J
authors.lastname: Lau
authors.forename: Amy L
authors.initials: AL
authors.lastname: Jackson
authors.forename: Jaime F
authors.initials: JF
authors.lastname: Modiano
authors.forename: Robert H
authors.initials: RH
authors.lastname: Weiss
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D002292:Carcinoma, Renal Cell / Q000201:enzymology* / Q000473:pathology*; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors*; D007680:Kidney Neoplasms / Q000201:enzymology* / Q000473:pathology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011687:Purines / Q000494:pharmacology*; D000077546:Roscovitine; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000187:drug effects* / Q000502:physiology*
chemical_list: D047428:Protein Kinase Inhibitors; D011687:Purines; D016159:Tumor Suppressor Protein p53; D000077546:Roscovitine; D018844:Cyclin-Dependent Kinases