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Induction of growth inhibition of 293 cells by downregulation of the cyclin E and cyclin-dependent kinase 4 proteins due to overexpression of TIS21.

Lim, IK; Lee, MS; Ryu, MS; Park, TJ; Fujiki, H; Eguchi, H; Paik, WK
Molecular carcinogenesis, 23(1):25-35, 1998
Journal Title
Molecular carcinogenesis
We earlier reported that TIS21 mRNA expression was markedly decreased in A549 and NCIH69 human lung cancer cells and in thymic carcinoma tissues obtained from transgenic mice containing simian virus 40 large T antigen (J Cancer Res Clin Oncol 121:279-284, 1995). To determine how TIS21 inhibits growth, we made 293 cells that constitutively expressed TIS21 protein. The constitutive TIS21 expresser lines C9 and C11 grew to a lower saturation density than did those in the vector-transfected clones (V7 and V10) and antisense-transfected clones (AS1 and AS4), and the size of the C9 and C11 cells increased significantly after transfection with TIS21 cDNA. The serum-stimulated cell cycle was analyzed by fluorescence-activated cell sorting after double thymidine treatment; V10 progressed normally through the cell division cycle, but C9 and C11 cells accumulated continuously in G1 phase until 36 h after treatment. On the other hand, the progression of cells that had already entered to S or G2/M phase was not inhibited. When cell-cycle regulatory proteins were measured, C9 and C11 cells showed significantly reduced synthesis of cyclin E and cyclin-dependent kinase (cdk) 4 as well as a decrease in cyclin E-associated cdk activity. These observations led us to conclude that TIS21 overexpression in G1 phase decreased the amounts of cyclin E and cdk4, thereby decreasing the activity of cdks at the G1-S transition.
MeSH terms
AnimalsBase SequenceCell Cycle Proteins/genetics/*physiologyCell Division/genetics/*physiologyCell LineCloning, MolecularCyclin E/*metabolismCyclin-Dependent Kinase 4Cyclin-Dependent Kinases/*metabolismDNA Primers*Down-Regulation*Genes, Tumor SuppressorHumansImmediate-Early Proteins/genetics/*physiologyMiceMice, Transgenic*Proto-Oncogene ProteinsRNA, Messenger/geneticsTumor Cells, CulturedTumor Suppressor Proteins
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Journal Papers > School of Medicine / Graduate School of Medicine > Biochemistry & Molecular Biology
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