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Involvement of p27Kip1 in G1 arrest by high dose 5α-dihydrotestosterone in LNCaP human prostate cancer cells

Oncogene volume 19, pages 670–679 (2000)Cite this article

Abstract

The cell cycle is governed by cyclin dependent kinases (cdks), which are activated by binding of cyclins, inhibited by cdk inhibitors and regulated by phosphorylation and dephosphorylation. Exposure to high dose dihydrotestosterone (DHT) inhibits population growth of the human prostate carcinoma cell line, LNCaP. To determine the mechanism of growth arrest by high dose DHT, we assayed the changes in cell cycle profile and the cell cycle regulators that mediate these effects. Treatment of asynchronously growing LNCaP cells with 100 nM DHT caused a G1 arrest. The proportion of cells in S phase fell from 22 to 2%, while the G1 fraction rose from 74 to 92% by 24 h. Loss of phosphorylation of the retinoblastoma protein was noted and cdk4 and cyclin E/cdk2 activities fell. Inhibition of these G1 cyclin dependent kinases was not due to loss of either cyclin or cdk proteins nor to increases in the cdk inhibitors p16INK4A and p21Cip1. p21Cip1 protein levels remained constant, and cyclin E-associated p21Cip1 fell, suggesting that p21Cip1 is not relevant to this form of cyclin E/cdk2 inhibition. Of note, total p27Kip1 levels and cyclin E-associated p27Kip1 increased as cells arrested and the amount of the CAK activated cdk2 bound to cyclin E decreased. p27Kip1 immunodepletion experiments demonstrated that the DHT-mediated increase in p27Kip1 was sufficient to fully saturate and inhibit target cyclin E/cdk2. The inhibition of cyclin E/cdk2 by p27Kip1 contributes to G1 arrest of LNCaP following high dose DHT. p27Kip1 may be a key effector of androgen dependent growth modulation in prostate cancer cells.

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Acknowledgements

We thank Jeff Donovan for helpful discussions and advice in the design of the antisense p27 experiments. This work was funded by a grant from the US Army Department of Defense Prostate Cancer Research Program to JM Slingerland. J Tsihlias was supported by a Terry Fox Fellowship from the National Cancer Institute of Canada. JM Slingerland is supported by Cancer Care Ontario.

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  1. M Flanagan

    Present address: Sunesis Pharmaceuticals, Redwood City, 94063, California, CA, USA

Authors and Affiliations

  1. Division of Cancer Research, Sunnybrook and Women's College Health Sciences Centre, Toronto, M4N 3M5, Ontario, Canada

    J Tsihlias, W Zhang, N Bhattacharya & J Slingerland

  2. Gilead Sciences, Foster City, 94404, California, CA, USA

    M Flanagan

  3. Division of Urology, Sunnybrook and Women's College Health Sciences Centre, Toronto, M4N 3M5, Ontario, Canada

    L Klotz

  4. Medical Oncology, Toronto-Sunnybrook Regional Cancer Centre, Ontario, Toronto, M4N 3M5, Canada

    J Slingerland

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Tsihlias, J., Zhang, W., Bhattacharya, N. et al. Involvement of p27Kip1 in G1 arrest by high dose 5α-dihydrotestosterone in LNCaP human prostate cancer cells. Oncogene 19, 670–679 (2000). https://doi.org/10.1038/sj.onc.1203369

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