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Hsp90-inhibitor geldanamycin abrogates G2 arrest in p53-negative leukemia cell lines through the depletion of Chk1

Oncogene volume 27, pages 3091–3101 (2008)Cite this article

Abstract

Checkpoint protein Chk1 has been identified as an Hsp90 client. Treatment with 100 nM geldanamycin (GM) for 24 h markedly reduced the Chk1 amount in Jurkat and ML-1 leukemia cell lines. Because Chk1 plays a central role in G2 checkpoint, we added GM to G2-arrested Jurkat and HL-60 cells pretreated with 50 nM doxorubicin for 24 h. GM slowly released both cell lines from doxorubicin-induced G2 arrest into G1 phase. GM also abrogated ICRF-193-induced decatenation G2 checkpoint in Jurkat and HL-60 cells. Western blot analysis showed that addition of GM attenuates doxorubicin- and ICRF-193-induced Chk1 phosphorylation at Ser345. GM, however, failed to abrogate G2 arrest in p53-positive ML-1 cells maybe due to the p21 induction. GM released HeLa cells from doxorubicin-induced G2 arrest but trapped them at M phase. Flow cytometric analysis showed that addition of GM converted doxorubicin-induced necrosis into apoptosis in Jurkat cells. Colony assay indicated that although GM has a weak cytotoxic effect as a single agent, it dramatically intensifies the cytotoxicity of doxorubicin and ICRF-193 in Jurkat and HL-60 cells. These results suggest that abrogation of G2 checkpoint by GM may play a central role in sensitizing p53-negative tumor cells to DNA-damaging and decatenation-inhibiting agents.

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Acknowledgements

This work was supported by grants from the Ministry of Education, Science, Sports and Culture in Japan. We thank Ms Keiko Hayashi and Ms Hiroko Hiraike for their excellent technical assistance.

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  1. Division of Hematology, Department of Internal Medicine, Juntendo University School of Medicine, Tokyo, Japan

    K Sugimoto, M Sasaki, Y Isobe, M Tsutsui, H Suto, J Ando, K Tamayose, M Ando & K Oshimi

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Correspondence to K Sugimoto.

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Sugimoto, K., Sasaki, M., Isobe, Y. et al. Hsp90-inhibitor geldanamycin abrogates G2 arrest in p53-negative leukemia cell lines through the depletion of Chk1. Oncogene 27, 3091–3101 (2008). https://doi.org/10.1038/sj.onc.1210978

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