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  • Original Paper
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Repression of NF-κB impairs HeLa cell proliferation by functional interference with cell cycle checkpoint regulators

Oncogene volume 18, pages 3213–3225 (1999)Cite this article

Abstract

NF-κB is an inducible transcription factor, which is regulated by interaction with inhibitory IκB proteins. Previous studies linked the activity of NF-κB to the proliferative state of the cell. Here we have analysed the function of NF-κB in the cell cycle. Inhibition of NF-κB in HeLa cells by stable overexpression of a transdominant negative IκB-α protein reduced cell growth. A kinetic analysis of the cell cycle revealed a retarded G1/S transition. The IκB-α overexpressing cell clones showed a decreased percentage of cells in the S phase and an impaired incorporation of bromodeoxyuridine (BrdU). The amounts of cyclins A, B1, D1, D3, and E were unchanged, but the G1-specific proteins cyclin D2 and cdk2 were strongly elevated in the IκB-α overexpressing cell clones. These cell clones also displayed an increase in cyclin D1-dependent kinase activity, pointing to a cell cycle arrest at the late G1 phase. IκB-α overexpression crosstalked to cell cycle checkpoints via a reduction of transcription factor p53 and elevation of p21WAF. Surprisingly, the IκB-α overexpressing cells showed an enrichment of c-Myc in the nucleoli, although the total amount of c-Myc protein was unchanged. These experiments identify an important contribution of the NF-κB/IκB system for the growth of HeLa cells.

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Abbreviations

BrdU:

bromodeoxyuridine

EMSA:

electrophoretic mobility shift assay

GST:

glutathione-S-transferase

IKK:

IκB-α kinase

IL-1:

interleukin-1

LPS:

lipopolysaccharide

TNF-α:

tumor necrosis factor α

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Acknowledgements

We thank Dr Dirk Eick (GSF, München) for the c-Myc expression vector, Dr Ed Harlow (Charlestown, MA, USA) for the GST-Rb expression vector, Dr Hubertus Stockinger (Boehringer Mannheim) for the monoclonal α-p65 antibody, Dr Ingrid Grummt (DKFZ, Heidelberg) for helpful discussions and Dr Peter Angel (DKFZ, Heidelberg) for antibodies recognizing cdk2, cyclin A and cyclin D2. The excellent technical assistance of Agnes Specking and Elmar Böhm is acknowledged. This work was supported in part by a European Biomed-2 grant (MLS), from the Volkswagen-Stiftung and Deutsche Forschungsgemeinschaft (CK), and the Cooperation Program in Cancer Research of the Deutsches Krebsforschungszentrum (DKFZ) and Israeli's Ministry of Science (SPH).

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  1. Barbara Kaltschmidt and Christian Kaltschmidt: B Kaltschmidt and C Kaltschmidt contributed equally to this paper

Authors and Affiliations

  1. Molecular Neurobiology Laboratory, Institute of Anatomy, Albert-Ludwigs-University, Freiburg, Stefan-Meier-Str. 19, Freiburg, D-79104, Germany

    Barbara Kaltschmidt & Christian Kaltschmidt

  2. Department of Immunochemistry, German Cancer Research Center, Im Neuenheimer Feld 280, 69120, Heidelberg, Germany

    Steffen P Hehner, Wulf Dröge & M Lienhard Schmitz

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Kaltschmidt, B., Kaltschmidt, C., Hehner, S. et al. Repression of NF-κB impairs HeLa cell proliferation by functional interference with cell cycle checkpoint regulators. Oncogene 18, 3213–3225 (1999). https://doi.org/10.1038/sj.onc.1202657

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