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[Cancer Research 61, 3986-3997, May 15, 2001]
© 2001 American Association for Cancer Research


Experimental Therapeutics

Akt/Protein Kinase B Is Constitutively Active in Non-Small Cell Lung Cancer Cells and Promotes Cellular Survival and Resistance to Chemotherapy and Radiation

John Brognard, Amy S. Clark, Yucheng Ni and Phillip A. Dennis1

Developmental Therapeutics Department, Medicine Branch, Division of Clinical Sciences, National Cancer Institute [J. B., A. S. C., Y. N.], and Navy Medical Oncology, National Naval Medical Center [P. A. D.], Bethesda, Maryland 20889

To evaluate the role of Akt/PKB in non-small cell lung cancer (NSCLC) survival, we analyzed NSCLC cell lines that differed in tumor histology as well as p53, Rb, and K-ras status. Constitutive Akt/protein kinase B (PKB) activity was demonstrated in 16 of 17 cell lines by maintenance of S473 phosphorylation with serum deprivation. Additional analysis of five of 2these NSCLC lines revealed that phosphorylation of S473 and T308 correlated with in vitro kinase activity. Akt/PKB activation was phosphatidylinositol 3-kinase-dependent and promoted survival because the phosphatidylinositol 3 inhibitors LY294002 and wortmannin inhibited Akt/PKB phosphorylation, Akt/PKB activity, and increased apoptosis only in cells with active Akt/PKB. To test whether Akt/PKB activity promoted therapeutic resistance, LY294002 was added with individual chemotherapeutic agents or irradiation. LY294002 greatly potentiated chemotherapy-induced apoptosis in cells with high Akt/PKB levels, but did not significantly increase chemotherapy-induced apoptosis in cells with low Akt/PKB levels. Combined with radiation in cells with active Akt/PKB, LY294002 additively increased apoptosis and inhibited clonogenic growth. These results were extended with transiently transfected Akt/PKB mutants. Transfecting dominant negative Akt/PKB decreased Akt/PKB activity and increased basal apoptosis as well as chemotherapy- and irradiation-induced apoptosis only in cells with high Akt/PKB activity. Conversely, transfecting constitutively active Akt/PKB into cells with low Akt/PKB activity increased Akt/PKB activity and attenuated chemotherapy- and radiation-induced apoptosis. We therefore identify Akt/PKB as a constitutively active kinase that promotes survival of NSCLC cells and demonstrate that modulation of Akt/PKB activity by pharmacological or genetic approaches alters the cellular responsiveness to therapeutic modalities typically used to treat patients with NSCLC.




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Cancer Res., December 1, 2004; 64(23): 8723 - 8730.
[Abstract] [Full Text] [PDF]


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D. J. VanderWeele, R. Zhou, and C. M. Rudin
Akt up-regulation increases resistance to microtubule-directed chemotherapeutic agents through mammalian target of rapamycin
Mol. Cancer Ther., December 1, 2004; 3(12): 1605 - 1613.
[Abstract] [Full Text] [PDF]