Genetic Inactivation of AKT1, AKT2, and PDPK1 in Human Colorectal Cancer Cells Clarifies Their Roles in Tumor Growth Regulation

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2010 Feb 6

PMID 20133737

Citations 80

Authors

Kajsa Ericson

Christine Gan

Ian Cheong

Carlo Rago

Yardena Samuels

Victor E Velculescu

Kenneth W Kinzler

David L Huso

Bert Vogelstein

Nickolas Papadopoulos

Affiliations

Soon will be listed here.

Abstract

Phosphotidylinositol-3-kinase (PI3K) signaling is altered in the majority of human cancers. To gain insight into the roles of members of this pathway in growth regulation, we inactivated AKT1, AKT2, or PDPK1 genes by targeted homologous recombination in human colon cancer cell lines. Knockout of either AKT1 or AKT2 had minimum effects on cell growth or downstream signaling. In contrast, knockout of both AKT1 and AKT2 resulted in markedly reduced proliferation in vitro when growth factors were limiting and severely affected experimental metastasis in mice. Unexpectedly, AKT1 and AKT2 appeared to regulate growth through FOXO proteins, but not through either GSK3beta or mTOR. In contrast, inactivation of PDPK1 affected GSK3beta and mTOR activation. These findings show that the PI3K signaling pathway is wired differently in human cancer cells than in other cell types or organisms, which has important implications for the design and testing of drugs that target this pathway.

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References

Vasudevan K, Barbie D, Davies M, Rabinovsky R, McNear C, Kim J . AKT-independent signaling downstream of oncogenic PIK3CA mutations in human cancer. Cancer Cell. 2009; 16(1):21-32. PMC: 2752826. DOI: 10.1016/j.ccr.2009.04.012. View

Hennessy B, Smith D, Ram P, Lu Y, Mills G . Exploiting the PI3K/AKT pathway for cancer drug discovery. Nat Rev Drug Discov. 2005; 4(12):988-1004. DOI: 10.1038/nrd1902. View

Hirata R, Chamberlain J, Dong R, Russell D . Targeted transgene insertion into human chromosomes by adeno-associated virus vectors. Nat Biotechnol. 2002; 20(7):735-8. DOI: 10.1038/nbt0702-735. View

Lawlor M, Mora A, Ashby P, Williams M, Murray-Tait V, Malone L . Essential role of PDK1 in regulating cell size and development in mice. EMBO J. 2002; 21(14):3728-38. PMC: 126129. DOI: 10.1093/emboj/cdf387. View

Vogt P, Bader A, Kang S . Phosphoinositide 3-kinase: from viral oncoprotein to drug target. Virology. 2005; 344(1):131-8. DOI: 10.1016/j.virol.2005.09.027. View

Kok K, Geering B, Vanhaesebroeck B . Regulation of phosphoinositide 3-kinase expression in health and disease. Trends Biochem Sci. 2009; 34(3):115-27. DOI: 10.1016/j.tibs.2009.01.003. View

Parsons D, Wang T, Samuels Y, Bardelli A, Cummins J, Delong L . Colorectal cancer: mutations in a signalling pathway. Nature. 2005; 436(7052):792. DOI: 10.1038/436792a. View

Katso R, Okkenhaug K, Ahmadi K, White S, Timms J, Waterfield M . Cellular function of phosphoinositide 3-kinases: implications for development, homeostasis, and cancer. Annu Rev Cell Dev Biol. 2001; 17:615-75. DOI: 10.1146/annurev.cellbio.17.1.615. View

Samuels Y, Diaz Jr L, Schmidt-Kittler O, Cummins J, Delong L, Cheong I . Mutant PIK3CA promotes cell growth and invasion of human cancer cells. Cancer Cell. 2005; 7(6):561-73. DOI: 10.1016/j.ccr.2005.05.014. View

10.

Wood L, Parsons D, Jones S, Lin J, Sjoblom T, Leary R . The genomic landscapes of human breast and colorectal cancers. Science. 2007; 318(5853):1108-13. DOI: 10.1126/science.1145720. View

11.

Staal S . Molecular cloning of the akt oncogene and its human homologues AKT1 and AKT2: amplification of AKT1 in a primary human gastric adenocarcinoma. Proc Natl Acad Sci U S A. 1987; 84(14):5034-7. PMC: 305241. DOI: 10.1073/pnas.84.14.5034. View

12.

Liu P, Cheng H, Roberts T, Zhao J . Targeting the phosphoinositide 3-kinase pathway in cancer. Nat Rev Drug Discov. 2009; 8(8):627-44. PMC: 3142564. DOI: 10.1038/nrd2926. View

13.

Bellacosa A, De Feo D, Godwin A, BELL D, Cheng J, Altomare D . Molecular alterations of the AKT2 oncogene in ovarian and breast carcinomas. Int J Cancer. 1995; 64(4):280-5. DOI: 10.1002/ijc.2910640412. View

14.

Rago C, Huso D, Diehl F, Karim B, Liu G, Papadopoulos N . Serial assessment of human tumor burdens in mice by the analysis of circulating DNA. Cancer Res. 2007; 67(19):9364-70. DOI: 10.1158/0008-5472.CAN-07-0605. View

15.

Rago C, Vogelstein B, Bunz F . Genetic knockouts and knockins in human somatic cells. Nat Protoc. 2007; 2(11):2734-46. DOI: 10.1038/nprot.2007.408. View

16.

Samuels Y, Velculescu V . Oncogenic mutations of PIK3CA in human cancers. Cell Cycle. 2004; 3(10):1221-4. DOI: 10.4161/cc.3.10.1164. View

17.

Grueneberg D, Degot S, Pearlberg J, Li W, Davies J, Baldwin A . Kinase requirements in human cells: I. Comparing kinase requirements across various cell types. Proc Natl Acad Sci U S A. 2008; 105(43):16472-7. PMC: 2575444. DOI: 10.1073/pnas.0808019105. View

18.

Whiteman E, Cho H, Birnbaum M . Role of Akt/protein kinase B in metabolism. Trends Endocrinol Metab. 2002; 13(10):444-51. DOI: 10.1016/s1043-2760(02)00662-8. View

19.

Chen W, Xu P, Gottlob K, Chen M, SOKOL K, Shiyanova T . Growth retardation and increased apoptosis in mice with homozygous disruption of the Akt1 gene. Genes Dev. 2001; 15(17):2203-8. PMC: 312770. DOI: 10.1101/gad.913901. View

20.

Vogt P, Gymnopoulos M, Hart J . PI 3-kinase and cancer: changing accents. Curr Opin Genet Dev. 2009; 19(1):12-7. PMC: 2746202. DOI: 10.1016/j.gde.2008.11.011. View