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Sensitized RNAi screen of human kinases and phosphatases identifies new regulators of apoptosis and chemoresistance

Nature Cell Biology volume 7, pages 591–600 (2005)Cite this article

Abstract

Evasion from apoptosis is a hallmark of cancer, and recent success using targeted therapeutics underscores the importance of identifying anti-apoptotic survival pathways. Here we utilize RNA interference (RNAi) to systematically screen the kinase and phosphatase component of the human genome. In addition to known kinases, we identified several new survival kinases. Interestingly, numerous phosphatases and associated regulatory subunits contribute to cell survival, revealing a previously unrecognized general role for phosphatases as negative regulators of apoptosis. We also identified a subset of phosphatases with tumour-suppressor-like activity. Finally, RNAi targeting of specific protein kinases sensitizes resistant cells to chemotherapeutic agents. The development of inhibitors that target these kinases or phosphatases may lead to new anti-cancer strategies.

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Figure 1: Human kinase siRNA library screen.
Figure 2: Human phosphatase siRNA library screen.
Figure 3: Human phosphatase siRNA library screen identifies cell death phosphatases and resistance to apoptosis.
Figure 4: Low-dose Taxol and siRNAs directed against survival kinases lead to enhanced apoptosis.
Figure 5: Synergistic chemotherapy-induced apoptosis in BT474 breast carcinoma cells with survival kinase siRNAs.

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Acknowledgements

We gratefully acknowledge E. Lader and S. Yerramilli for their assistance with siRNA library design and QRT–PCR expertise. We thank F. Tsai for help with data analysis, W. Dowdle for technical assistance, and C. Dimitri, B. Ballif and members of the Blenis laboratory for their critical feedback. This work is supported by National Institutes of Health grants RO1CA46595 and GM51405 to J.B. J.M. is a Fellow of the American Cancer Society and L.M. is a Special Fellow of the Leukemia & Lymphoma Society.

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  1. Jeffrey P. MacKeigan, Leon O. Murphy and John Blenis: These authors contributed equally to this work.

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  1. Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, 02115, Massachusetts, USA

    Jeffrey P. MacKeigan, Leon O. Murphy & John Blenis

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  1. Jeffrey P. MacKeigan
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Correspondence to John Blenis.

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MacKeigan, J., Murphy, L. & Blenis, J. Sensitized RNAi screen of human kinases and phosphatases identifies new regulators of apoptosis and chemoresistance. Nat Cell Biol 7, 591–600 (2005). https://doi.org/10.1038/ncb1258

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