PDK 1 potentiates upstream lesions on the PI 3 K pathway in breast carcinoma

Lesions of ERBB2, PTEN, and PIK3CA activate the PI3K pathway during cancer development by increasing levels of phosphatidylinositol-3,4,5-triphosphate (PIP3). 3-phosphoinositide-dependent kinase 1 (PDK1) is the first node of the PI3K signal output and is required for activation of AKT. PIP3 recruits PDK1 and AKT to the cell membrane through interactions with their PH domains, allowing PDK1 to activate AKT by phosphorylating it at residue threonine 308. We show that total PDK1 protein and mRNA was over-expressed in a majority of human breast cancers and that 21% of tumors had five or more copies of the gene encoding PDK1, PDPK1. We found that increased PDPK1 copy number was associated with upstream pathway lesions (ERBB2 amplification, PTEN loss, or PIK3CA mutation), as well as patient survival. Examination of an independent set of breast cancers and tumor cell lines derived from multiple forms of human cancers also found increased PDK1 protein levels associated with such upstream pathway lesions. In human mammary cells, PDK1 enhanced the ability of upstream lesions to signal to AKT, stimulate cell growth and migration, and rendered cells more resistant to PDK1 and PI3K inhibition. After orthotopic transplantation, PDK1 overexpression was not oncogenic but dramatically enhanced the ability of ERBB2 to form tumors. Our studies argue that PDK1 overexpression and increased PDPK1 copy number are common occurrences in cancer that potentiate the oncogenic effect of upstream lesions on the PI3K Requests for reprints: Ramon Parsons, Institute for Cancer Genetics, Columbia University, ICRC407A, 1130 Saint Nicholas Avenue, New York, NY 10032. Phone 212-851-5278. Fax 212-851-5256. Email rep15@columbia.edu. Note: Supplementary data are available at CancerResearch Online (http://cancerres.aacrjournals.org/). Disclosure of Potential Conflicts of Interest No potential conflicts of interest were disclosed. NIH Public Access Author Manuscript Cancer Res. Author manuscript; available in PMC 2010 August 1. Published in final edited form as: Cancer Res. 2009 August 1; 69(15): 6299–6306. doi:10.1158/0008-5472.CAN-09-0820. N IH PA Athor M anscript N IH PA Athor M anscript N IH PA Athor M anscript pathway. Therefore, we conclude that alteration of PDK1 is a critical component of oncogenic PI3K signaling in breast cancer.