Assay for Isolation of Inhibitors of Her2-Kinase Expression
Her2 (ErbB2) protein is overexpressed in breast and other solid tumors, and its expression is associated with progressive disease. Current therapies directed toward Her2 either block dimerization of the receptor or inhibit tyrosine kinase activity to disrupt intracellular signaling. However, little is known about alternative mechanisms for suppressing Her2 expression, possibly by inducing degradation or blocking synthesis. Here, we describe a hybrid western-blotting and enzyme-linked immunosorbent assay (ELISA) designed to identify in low- to medium-throughput format noncytotoxic compounds that reduce expression of Her2 protein.
Key wordsBreast cancer Heat-shock protein 90 (Hsp90) Her2 (ErbB2) tyrosine kinase Whole-cell immunoblot
The author would like to thank Joungnam Kim and Cristina C. Clement for technical assistance. This work was supported by R03 MH076408, and a Susan G. Komen grant, R01-SGK-BCTR0504381.
- 7.Citri, A., Kochupurakkal, B. S., and Yarden, Y. (2004) The achilles heel of ErbB-2/HER2: regulation by the Hsp90 chaperone machine and potential for pharmacological intervention. Cell Cycle 1, 51–60.Google Scholar
- 14.Haggarty, S. J., Mayer, T. U., Miyamoto, D. T., Fathi, R., King, R. W., Mitchison, T. J., and Schreiber, S. L. (2000) Dissecting cellular processes using small molecules: identification of colchicine-like, taxol-like and other small molecules that perturb mitosis. Chem. Biol. 4, 275–286.CrossRefGoogle Scholar
- 15.Blaskovich, M. A., Sun, J., Cantor, A., Turkson, J., Jove, R., and Sebti, S. M. (2003) Discovery of JSI-124 (cucurbitacin I), a selective Janus kinase/signal transducer and activator of transcription 3 signaling pathway inhibitor with potent antitumor activity against human and murine cancer cells in mice. Cancer Res. 63, 1270–1279.PubMedGoogle Scholar