title: Cell cycle inhibitors (p27Kip1 and p21CIP1) cause hypertrophy in LLC-PK1 cells.
abstract: BACKGROUND
  Angiotensin II has been reported to induce renal tubular hypertrophy, but the mechanisms of this hypertrophy are not well known. We evaluated the roles of cyclin-dependent kinase (CDK) inhibitors in renal tubular hypertrophy.
  
  
  METHODS
  To elucidate whether CDK inhibitors cause renal tubular hypertrophy, we produced adenovirus vectors containing coding sequences of the CDK inhibitors p27Kip1 (AxCAp27), p21CIP1 (AxCAp21), and p16INK4 (AxCAp16), and we investigated the effect of these gene transfers on epidermal growth factor (EGF)-induced proliferation in LLC-PK1 cells. We evaluated the cell cycle and hypertrophy by measurements of the [3H]-leucine and [3H]-thymidine incorporation, the protein:DNA ratio, flow cytometry, and CDK4 and CDK2 kinase assays.
  
  
  RESULTS
  AxCAp27 and AxCAp21 caused significant increases in [3H]-leucine incorporation and the protein:DNA ratio but did not change the [3H]-thymidine incorporation. Conversely, AxCAp16 inhibited EGF-stimulated [3H]-thymidine incorporation but did not change the [3H]-leucine incorporation. AxCAp27, AxCAp21, and AxCAp16 all inhibited EGF-stimulated CDK4 kinase activity (to 15.6, 14.1, and 21.9% of control, respectively). Forward light-scatter analysis demonstrated that AxCAp27 and AxCAp21 increased the cell size but that AxCAp16 effected no change in cell size.
  
  
  CONCLUSIONS
  These findings suggest that p27Kip1 and p21CIP1 may play an important role in hypertrophy of renal tubule cells by reducing pRb phosphorylation. On the other hand, p16INK4 was not found to cause hypertrophic changes in EGF-treated LLC-PK1 cells.
authors.affiliation: Second Department of Internal Medicine, Tokyo Medical and Dental University, Japan. yterada.med2@med.tmd.ac.jp
authors.forename: Y
authors.initials: Y
authors.lastname: Terada
authors.forename: S
authors.initials: S
authors.lastname: Inoshita
authors.forename: O
authors.initials: O
authors.lastname: Nakashima
authors.forename: M
authors.initials: M
authors.lastname: Tamamori
authors.forename: H
authors.initials: H
authors.lastname: Ito
authors.forename: M
authors.initials: M
authors.lastname: Kuwahara
authors.forename: S
authors.initials: S
authors.lastname: Sasaki
authors.forename: F
authors.initials: F
authors.lastname: Marumo
mesh_terms: D000256:Adenoviridae; D000804:Angiotensin II / Q000494:pharmacology; D000818:Animals; D042846:CDC2-CDC28 Kinases; D002352:Carrier Proteins / Q000235:genetics; D018797:Cell Cycle Proteins; D048429:Cell Size; D051357:Cyclin-Dependent Kinase 2; D051358:Cyclin-Dependent Kinase 4; D050762:Cyclin-Dependent Kinase Inhibitor p15; D019941:Cyclin-Dependent Kinase Inhibitor p16; D050759:Cyclin-Dependent Kinase Inhibitor p21; D050760:Cyclin-Dependent Kinase Inhibitor p27; D018844:Cyclin-Dependent Kinases / Q000378:metabolism; D016213:Cyclins / Q000235:genetics*; D004791:Enzyme Inhibitors / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D006984:Hypertrophy; D007674:Kidney Diseases / Q000201:enzymology / Q000473:pathology; D018374:LLC-PK1 Cells / Q000187:drug effects / Q000201:enzymology / Q000473:pathology*; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins; D013552:Swine; D014316:Tritium; D025521:Tumor Suppressor Proteins; D014662:Vasoconstrictor Agents / Q000494:pharmacology; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D050762:Cyclin-Dependent Kinase Inhibitor p15; D019941:Cyclin-Dependent Kinase Inhibitor p16; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D004791:Enzyme Inhibitors; D008869:Microtubule-Associated Proteins; D011518:Proto-Oncogene Proteins; D025521:Tumor Suppressor Proteins; D014662:Vasoconstrictor Agents; D014316:Tritium; D000804:Angiotensin II; D050760:Cyclin-Dependent Kinase Inhibitor p27; D017346:Protein Serine-Threonine Kinases; D042846:CDC2-CDC28 Kinases; D051357:Cyclin-Dependent Kinase 2; D051358:Cyclin-Dependent Kinase 4; D018844:Cyclin-Dependent Kinases; D001616:beta-Galactosidase