Abstract
Class IA phosphoinositide-3 kinases (PI3Ks) signaling has recently emerged as a key element in cancer development because of its ability to trigger a complex panoply of cellular responses controlling survival and proliferation. Many cancers show inappropriately activated PI3K pathway, and tumors with high PI3K activity are frequently resistant to traditional chemotherapy. Indeed, preclinical studies demonstrated a prominent role for the PI3K pathway in cancer cell survival and growth, thus validating PI3K as a potential drug target in cancer. The emerging interest in inhibiting PI3Ks in cancer have prompted the aggressive development of new selective PI3K pathway inhibitors as cancer therapy, and many of these molecules are currently in early-phase clinical trials. In this chapter, we describe methods to measure the PI3K lipid kinase activity in vitro, which is the standard procedure to test the efficacy of inhibitors.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Engelman, J. A., Luo, J., and Cantley, L. C. (2006) The evolution of phosphatidylinositol 3-kinases as regulators of growth and metabolism. Nat. Rev. Genet. 7, 606–619.
Hirsch, E., Ciraolo, E., Ghigo, A., and Costa, C. (2008) Taming the PI3K team to hold inflammation and cancer at bay, Pharmacology & therapeutics 118, 192–205.
Nadal, E., and Olavarria, E. (2004) Imatinib mesylate (Gleevec/Glivec) a molecular-targeted therapy for chronic myeloid leukaemia and other malignancies. International journal of clinical practice 58, 511–516.
Helfrich, B. A., Raben, D., Varella-Garcia, M., Gustafson, D., Chan, D. C., Bemis, L., Coldren, C., Baron, A., Zeng, C., Franklin, W. A., Hirsch, F. R., Gazdar, A., Minna, J., and Bunn, P. A., Jr. (2006) Antitumor activity of the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor gefitinib (ZD1839, Iressa) in non-small cell lung cancer cell lines correlates with gene copy number and EGFR mutations but not EGFR protein levels. Clin. Cancer Res. 12, 7117–7125.
McCubrey, J. A., Steelman, L. S., Abrams, S. L., Lee, J. T., Chang, F., Bertrand, F. E., Navolanic, P. M., Terrian, D. M., Franklin, R. A., D’Assoro, A. B., Salisbury, J. L., Mazzarino, M. C., Stivala, F., and Libra, M. (2006) Roles of the RAF/MEK/ERK and PI3K/PTEN/AKT pathways in malignant transformation and drug resistance. Advances in enzyme regulation 46, 249–279.
Samuels, Y., Wang, Z., Bardelli, A., Silliman, N., Ptak, J., Szabo, S., Yan, H., Gazdar, A., Powell, S. M., Riggins, G. J., Willson, J. K., Markowitz, S., Kinzler, K. W., Vogelstein, B., and Velculescu, V. E. (2004) High frequency of mutations of the PIK3CA gene in human Âcancers. Science 304, 554.
Engelman, J. A. (2009) Targeting PI3K signalling in cancer: opportunities, challenges and limitations. Nat. Rev. Cancer 9, 550–562.
Geering, B., Cutillas, P. R., Nock, G., Gharbi, S. I., and Vanhaesebroeck, B. (2007) Class IA phosphoinositide 3-kinases are obligate p85-p110 heterodimers. Proc. Natl. Acad. Sci.USA 104, 7809–7814.
Vanhaesebroeck, B., and Waterfield, M. D. (1999) Signaling by distinct classes of phosphoinositide 3-kinases. Exp. Cell Res. 253, 239–254.
Kahn, C. R., White, M. F., Shoelson, S. E., Backer, J. M., Araki, E., Cheatham, B., Csermely, P., Folli, F., Goldstein, B. J., Huertas, P., and et al. (1993) The insulin receptor and its substrate: molecular determinants of early events in insulin action. Rec. Prog. Horm. Res. 48, 291–339.
Beeton, C. A., Chance, E. M., Foukas, L. C., and Shepherd, P. R. (2000) Comparison of the kinetic properties of the lipid- and protein-kinase activities of the p110alpha and p110beta catalytic subunits of class-Ia phosphoinositide 3-kinases. Biochem J.350 Pt 2, 353–359.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Ciraolo, E., Perino, A., Hirsch, E. (2012). Measuring PI3K Lipid Kinase Activity. In: Kuster, B. (eds) Kinase Inhibitors. Methods in Molecular Biology, vol 795. Humana Press. https://doi.org/10.1007/978-1-61779-337-0_4
Download citation
DOI: https://doi.org/10.1007/978-1-61779-337-0_4
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-336-3
Online ISBN: 978-1-61779-337-0
eBook Packages: Springer Protocols