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[Cancer Research 59, 4634-4641, September 1, 1999]
© 1999 American Association for Cancer Research
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[Cancer Research 59, 4634-4641, September 15, 1999]
© 1999 American Association for Cancer Research

Experimental Therapeutics

Down-Regulation of Cyclin D1 by Transcriptional Repression in MCF-7 Human Breast Carcinoma Cells Induced by Flavopiridol1

Brad Carlson, Tyler Lahusen, Sheo Singh, Andrea Loaiza-Perez, Peter J. Worland, Richard Pestell, Chris Albanese, Edward A. Sausville and Adrian M. Senderowicz2

Developmental Therapeutics Program (DTP), DTP Clinical Trials Unit, Division of Cancer Treatment and Diagnosis, National Cancer Institute, Bethesda, Maryland 20892 [B. C., T. L. S. S., A. L-P., E. A. S., A. M. S.]; Department of Medicine and Developmental and Molecular Biology, The Albert Einstein Cancer Center, Bronx, New York 10461 [R. P., C. A.]; and Mitotix Inc., Cambridge, Massachusetts 02139 [P. J. W.]

Flavopiridol is a novel flavonoid that induces cell cycle arrest at different stages of the cell cycle because of the inhibition of cyclin-dependent kinases (cdks). In previous studies from our laboratory, (B. A. Carlson et al., Cancer Res., 56: 2973–2978, 1996), we observed that exposure of the MCF-7 breast carcinoma cell line to flavopiridol resulted in G1-S arrest, which was associated with the loss of cdk4 and cdk2 activity by 24 h of exposure. Along with this inhibition, flavopiridol decreased total cyclin-D protein levels in this cell line. In this work, we demonstrate that using isoform-specific antibodies, flavopiridol induces an early (by 6 h) decrease in cyclin D1 protein levels. This decline is followed by a decline in cyclin D3 with no effect on cyclin D2 or cyclin E levels by 10 h. Furthermore, at early time points (up to 8 h), the activity of cdk4 and the expression of endogenous phosphorylated retinoblastoma species from intact cells exposed to flavopiridol are unchanged. Thus, the decline in cdk4 activity and the induction of retinoblastoma hypophosphorylation follows cyclin D1 decline. Turnover studies demonstrate that the half-life of cyclin D1 (~30 min) is not shortened in flavopiridol-exposed cells, and that the turnover of cdk4-bound cyclin D1 is unaltered. However, steady-state levels of cyclin D1 mRNA display a significant decrease by 4 h of flavopiridol treatment, with total disappearance by 8 h. This mRNA decline is not abrogated by the presence of cycloheximide. Furthermore, we have found that flavopiridol specifically represses the activity of the full-length cyclin D1 promoter linked to a luciferase reporter gene. In summary, we have found that the flavopiridol-induced decline in cyclin D1 is an early event, specific and, at least in part, due to the transcriptional repression of the cyclin D1 promoter. These results extend our understanding of flavopiridol’s action to include regulation of cyclin D1 transcription.




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GL331 Induces Down-Regulation of Cyclin D1 Expression via Enhanced Proteolysis and Repressed Transcription
Mol. Pharmacol., October 1, 2001; 60(4): 768 - 775.
[Abstract] [Full Text] [PDF]

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 Cancer Res.Home page
M. E. Lane, B. Yu, A. Rice, K. E. Lipson, C. Liang, L. Sun, C. Tang, G. McMahon, R. G. Pestell, and S. Wadler
A Novel cdk2-selective Inhibitor, SU9516, Induces Apoptosis in Colon Carcinoma Cells
Cancer Res., August 1, 2001; 61(16): 6170 - 6177.
[Abstract] [Full Text] [PDF]

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 Clin. Cancer Res.Home page
C. P. Jung, M. V. Motwani, and G. K. Schwartz
Flavopiridol Increases Sensitization to Gemcitabine in Human Gastrointestinal Cancer Cell Lines and Correlates with Down-Regulation of Ribonucleotide Reductase M2 Subunit
Clin. Cancer Res., August 1, 2001; 7(8): 2527 - 2536.
[Abstract] [Full Text] [PDF]

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 Clin. Cancer Res.Home page
G. I. Shapiro, J. G. Supko, A. Patterson, C. Lynch, J. Lucca, P. F. Zacarola, A. Muzikansky, J. J. Wright, T. J. Lynch Jr., and B. J. Rollins
A Phase II Trial of the Cyclin-dependent Kinase Inhibitor Flavopiridol in Patients with Previously Untreated Stage IV Non-Small Cell Lung Cancer
Clin. Cancer Res., June 1, 2001; 7(6): 1590 - 1599.
[Abstract] [Full Text] [PDF]

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 JCOHome page
E. A. Sausville, S. G. Arbuck, R. Messmann, D. Headlee, K. S. Bauer, R. M. Lush, A. Murgo, W. D. Figg, T. Lahusen, S. Jaken, et al.
Phase I Trial of 72-Hour Continuous Infusion UCN-01 in Patients With Refractory Neoplasms
J. Clin. Oncol., April 15, 2001; 19(8): 2319 - 2333.
[Abstract] [Full Text] [PDF]

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 Mol. Pharmacol.Home page
X.-K. Li, M. Motwani, W. Tong, W. Bornmann, and G. K. Schwartz
Huanglian, A Chinese Herbal Extract, Inhibits Cell Growth by Suppressing the Expression of Cyclin B1 and Inhibiting CDC2 Kinase Activity in Human Cancer Cells
Mol. Pharmacol., April 13, 2001; 58(6): 1287 - 1293.
[Abstract] [Full Text]

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 JCOHome page
G. K. Schwartz, D. Ilson, L. Saltz, E. O'Reilly, W. Tong, P. Maslak, J. Werner, P. Perkins, M. Stoltz, and D. Kelsen
Phase II Study of the Cyclin-Dependent Kinase Inhibitor Flavopiridol Administered to Patients With Advanced Gastric Carcinoma
J. Clin. Oncol., April 1, 2001; 19(7): 1985 - 1992.
[Abstract] [Full Text] [PDF]

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 Cancer Res.Home page
W. Li, J. Fan, and J. R. Bertino
Selective Sensitization of Retinoblastoma Protein-deficient Sarcoma Cells to Doxorubicin by Flavopiridol-mediated Inhibition of Cyclin-dependent Kinase 2 Kinase Activity
Cancer Res., March 1, 2001; 61(6): 2579 - 2582.
[Abstract] [Full Text]

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 Cancer Res.Home page
L. Cartee, Z. Wang, R. H. Decker, S. P. Chellappan, G. Fusaro, K. G. Hirsch, H. M. Sankala, P. Dent, and S. Grant
The Cyclin-dependent Kinase Inhibitor (CDKI) Flavopiridol Disrupts Phorbol 12-Myristate 13-Acetate-induced Differentiation and CDKI Expression while Enhancing Apoptosis in Human Myeloid Leukemia Cells
Cancer Res., March 1, 2001; 61(6): 2583 - 2591.
[Abstract] [Full Text]

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 JNCI J Natl Cancer InstHome page
A. M. Senderowicz and E. A. Sausville
Preclinical and Clinical Development of Cyclin-Dependent Kinase Modulators
J Natl Cancer Inst, March 1, 2000; 92(5): 376 - 387.
[Abstract] [Full Text] [PDF]

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 JNCI J Natl Cancer InstHome page
S. M. Lippman and P. H. Brown
Tamoxifen Prevention of Breast Cancer: an Instance of the Fingerpost
J Natl Cancer Inst, November 3, 1999; 91(21): 1809 - 1819.
[Full Text] [PDF]

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 J. Biol. Chem.Home page
S.-H. Chao, K. Fujinaga, J. E. Marion, R. Taube, E. A. Sausville, A. M. Senderowicz, B. M. Peterlin, and D. H. Price
Flavopiridol Inhibits P-TEFb and Blocks HIV-1 Replication
J. Biol. Chem., September 8, 2000; 275(37): 28345 - 28348.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
S.-H. Chao and D. H. Price
Flavopiridol Inactivates P-TEFb and Blocks Most RNA Polymerase II Transcription in Vivo
J. Biol. Chem., August 17, 2001; 276(34): 31793 - 31799.
[Abstract] [Full Text] [PDF]


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