The protein kinase Akt induces epithelial mesenchymal transition and promotes enhanced motility and invasiveness of squamous cell carcinoma lines.

Epithelial-mesenchymal transition (EMT) is an important process during development and oncogenesis by which epithelial cells acquire fibroblast-like properties and show reduced intercellular adhesion and increased motility. Squamous cell carcinoma lines engineered to express constitutively active Akt underwent EMT, characterized by down-regulation of the epithelial markers desmoplakin, E-cadherin, and beta-catenin and up-regulation of the mesenchymal marker vimentin. The cells lost epithelial cell morphology and acquired fibroblast-like properties. Additionally, E-cadherin was down-regulated transcriptionally. The cells expressing constitutively active Akt exhibited reduced cell-cell adhesion, increased motility on fibronectin-coated surfaces, and increased invasiveness in animals. AKT is activated in many human carcinomas, and the AKT-driven EMT may confer the motility required for tissue invasion and metastasis. These findings suggest that future therapies based on AKT inhibition may complement conventional treatments by controlling tumor cell invasion and metastasis.

[1]  G. Panayotou,et al.  Small GTPases and tyrosine kinases coregulate a molecular switch in the phosphoinositide 3-kinase regulatory subunit. , 2002, Cancer cell.

[2]  Y. Ip,et al.  Snail/slug family of repressors: slowly going into the fast lane of development and cancer. , 2000, Gene.

[3]  Randall T Moon,et al.  Mechanism and function of signal transduction by the Wnt/β-catenin and Wnt/Ca2+ pathways , 1999, Oncogene.

[4]  D. Hanahan,et al.  Elevated levels of IGF-1 receptor convey invasive and metastatic capability in a mouse model of pancreatic islet tumorigenesis. , 2002, Cancer cell.

[5]  Kenneth M. Yamada,et al.  Transmembrane crosstalk between the extracellular matrix and the cytoskeleton , 2001, Nature Reviews Molecular Cell Biology.

[6]  D. Littman,et al.  Packaging system for rapid production of murine leukemia virus vectors with variable tropism , 1992, Journal of virology.

[7]  P. Cohen,et al.  Inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B , 1995, Nature.

[8]  Michael Kühl,et al.  Functional interaction of β-catenin with the transcription factor LEF-1 , 1996, Nature.

[9]  A. G. Herreros,et al.  The transcription factor Snail is a repressor of E-cadherin gene expression in epithelial tumour cells , 2000, Nature Cell Biology.

[10]  C. Heldin,et al.  TGF-β type I receptor / ALK-5 and Smad proteins mediate epithelial to mesenchymal transdifferentiation in NMuMG breast epithelial cells , 1999 .

[11]  W. Fiers,et al.  Genetic manipulation of E-cadherin expression by epithelial tumor cells reveals an invasion suppressor role , 1991, Cell.

[12]  J. Cheng,et al.  Molecular alterations of the AKT2 oncogene in ovarian and breast carcinomas , 1995, International journal of cancer.

[13]  S. Y. Zhang,et al.  p53 alterations in human squamous cell carcinomas and carcinoma cell lines. , 1993, The American journal of pathology.

[14]  J. Testa,et al.  AKT activation up-regulates insulin-like growth factor I receptor expression and promotes invasiveness of human pancreatic cancer cells. , 2001, Cancer research.

[15]  F. McCormick,et al.  Wnt Signaling to β-Catenin Involves Two Interactive Components , 2000, The Journal of Biological Chemistry.

[16]  J. Cheng,et al.  Amplification of AKT2 in human pancreatic cells and inhibition of AKT2 expression and tumorigenicity by antisense RNA. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[17]  B. Boyer,et al.  Induction and regulation of epithelial-mesenchymal transitions. , 2000, Biochemical pharmacology.

[18]  L. Liotta,et al.  General mechanisms of metastasis , 1997, Cancer.

[19]  P. Polakis.  Wnt signaling and cancer. , 2000, Genes & development.

[20]  Domenico Coppola,et al.  Frequent activation of AKT2 and induction of apoptosis by inhibition of phosphoinositide-3-OH kinase/Akt pathway in human ovarian cancer , 2000, Oncogene.

[21]  Kenneth M. Yamada,et al.  Fibronectin, integrins, and growth control , 2001, Journal of cellular physiology.

[22]  C. Heldin,et al.  TGF-(beta) type I receptor/ALK-5 and Smad proteins mediate epithelial to mesenchymal transdifferentiation in NMuMG breast epithelial cells. , 1999, Journal of cell science.

[23]  J E Paciga,et al.  AKT1/PKBalpha kinase is frequently elevated in human cancers and its constitutive activation is required for oncogenic transformation in NIH3T3 cells. , 2001, The American journal of pathology.

[24]  P. Cohen,et al.  A common phosphate binding site explains the unique substrate specificity of GSK3 and its inactivation by phosphorylation. , 2001, Molecular Cell.

[25]  P. Dennis,et al.  Akt/protein kinase B is constitutively active in non-small cell lung cancer cells and promotes cellular survival and resistance to chemotherapy and radiation. , 2001, Cancer research.

[26]  F. McCormick,et al.  Differential Regulation of Glycogen Synthase Kinase 3β by Insulin and Wnt Signaling* , 2000, The Journal of Biological Chemistry.

[27]  W. Birchmeier,et al.  Dissecting tumor cell invasion: epithelial cells acquire invasive properties after the loss of uvomorulin-mediated cell-cell adhesion , 1989, The Journal of cell biology.

[28]  Miller Ad,et al.  Improved Retroviral Vectors for Gene Transfer and Expression , 1989 .

[29]  Walter Birchmeier,et al.  Hot papers in cell biology - J. Behrens, J.P. von Kries, M. Kuehl, L. Bruhn, D. Wedlich, R. Grosschedl, W. Birchmeier: "Functional interaction of beta-catenin with the transcription factor LEF-1" - Comments by Walter Birchmeier , 1999 .

[30]  C. Thompson,et al.  Cell metabolism in the regulation of programmed cell death , 2002, Trends in Endocrinology & Metabolism.

[31]  H. Nakayama,et al.  High expression levels of nuclear factor κB, IκB kinase α and Akt kinase in squamous cell carcinoma of the oral cavity , 2001 .

[32]  J. Cheng,et al.  AKT2, a putative oncogene encoding a member of a subfamily of protein-serine/threonine kinases, is amplified in human ovarian carcinomas. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[33]  J. Jouanneau,et al.  Expression of transfected transforming growth factor alpha induces a motile fibroblast-like phenotype with extracellular matrix-degrading potential in a rat bladder carcinoma cell line. , 1990, Cell regulation.

[34]  S. Fukumoto,et al.  Akt Participation in the Wnt Signaling Pathway through Dishevelled* , 2001, The Journal of Biological Chemistry.

[35]  B. Herrmann,et al.  Nuclear localization of β-catenin by interaction with transcription factor LEF-1 , 1996, Mechanisms of Development.

[36]  J. Thiery,et al.  Phosphorylation of Tyrosine Residues 31 and 118 on Paxillin Regulates Cell Migration through an Association with Crk in Nbt-II Cells , 2000, The Journal of cell biology.

[37]  R. Nusse,et al.  WNT targets. Repression and activation. , 1999, Trends in genetics : TIG.

[38]  W. Birchmeier,et al.  E-cadherin as an invasion suppressor. , 1995, Ciba Foundation symposium.

[39]  C. Weston,et al.  Signal transduction: signaling specificity- a complex affair. , 2001, Science.

[40]  D. Barritault,et al.  Acidic fibroblast growth factor is a modulator of epithelial plasticity in a rat bladder carcinoma cell line. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[41]  Alfonso Bellacosa,et al.  AKT plays a central role in tumorigenesis , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[42]  P. Woodman.  Biogenesis of the Sorting Endosome: The Role of Rab5 , 2000, Traffic.

[43]  Walter G. Gunn.  Cancer: Principles and Practice of Oncology , 1982 .

[44]  M. Perricaudet,et al.  Widespread long-term gene transfer to mouse skeletal muscles and heart. , 1992, The Journal of clinical investigation.

[45]  R Kemler,et al.  A role for cadherins in tissue formation. , 1996, Development.

[46]  G. Berx,et al.  The two-handed E box binding zinc finger protein SIP1 downregulates E-cadherin and induces invasion. , 2001, Molecular cell.

[47]  M. Saji,et al.  Overexpression and overactivation of Akt in thyroid carcinoma. , 2001, Cancer research.

[48]  A. Klein-Szanto,et al.  Elevated furin expression in aggressive human head and neck tumors and tumor cell lines , 2001, Molecular carcinogenesis.

[49]  Véronique Delmas,et al.  IGF-II induces rapid β-catenin relocation to the nucleus during epithelium to mesenchyme transition , 2001, Oncogene.

[50]  M. Peifer,et al.  Abelson kinase regulates epithelial morphogenesis in Drosophila , 2001, The Journal of cell biology.

[51]  A. Klein-Szanto,et al.  Furin inhibition results in absent or decreased invasiveness and tumorigenicity of human cancer cells , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[52]  M. Nieto,et al.  The snail superfamily of zinc-finger transcription factors , 2002, Nature Reviews Molecular Cell Biology.

[53]  J. Thiery,et al.  Expression of the cytoplasmic domain of E-cadherin induces precocious mammary epithelial alveolar formation and affects cell polarity and cell-matrix integrity. , 1999, Developmental biology.

[54]  Laurence H. Pearl,et al.  Crystal Structure of Glycogen Synthase Kinase 3β Structural Basis for Phosphate-Primed Substrate Specificity and Autoinhibition , 2001, Cell.

[55]  R. Roth,et al.  Protein Kinase B/akt and Rab5 Mediate Ras Activation of Endocytosis* , 1998, The Journal of Biological Chemistry.

[56]  B. Geiger,et al.  Transmembrane crosstalk between the extracellular matrix--cytoskeleton crosstalk. , 2001, Nature reviews. Molecular cell biology.

[57]  M. Cavet,et al.  Constitutively Active Phosphatidylinositol 3-Kinase and AKT Are Sufficient to Stimulate the Epithelial Na+/H+ Exchanger 3* , 2001, The Journal of Biological Chemistry.

[58]  R. Glazer,et al.  Akt1 induces extracellular matrix invasion and matrix metalloproteinase-2 activity in mouse mammary epithelial cells. , 2001, Cancer research.

[59]  A. Miller,et al.  Improved retroviral vectors for gene transfer and expression. , 1989, BioTechniques.

[60]  J. Testa,et al.  Structure, expression and chromosomal mapping of c-akt: relationship to v-akt and its implications. , 1993, Oncogene.

[61]  J. Thiery,et al.  IGF-II promotes mesoderm formation. , 2000, Developmental biology.

[62]  V. Devita,et al.  Cancer : Principles and Practice of Oncology , 1982 .

[63]  H. Moses,et al.  Phosphatidylinositol 3-Kinase Function Is Required for Transforming Growth Factor β-mediated Epithelial to Mesenchymal Transition and Cell Migration* , 2000, The Journal of Biological Chemistry.

[64]  A. García-Pardo,et al.  Activation of the alpha 4 beta 1 integrin through the beta 1 subunit induces recognition of the RGDS sequence in fibronectin , 1994, The Journal of cell biology.

[65]  J. Testa,et al.  The proteins encoded by c-akt and v-akt differ in post-translational modification, subcellular localization and oncogenic potential. , 1993, Oncogene.

[66]  Francisco Portillo,et al.  The transcription factor Snail controls epithelial–mesenchymal transitions by repressing E-cadherin expression , 2000, Nature Cell Biology.

[67]  F. Kikkawa,et al.  Fibronectin activates matrix metalloproteinase-9 secretion via the MEK1-MAPK and the PI3K-Akt pathways in ovarian cancer cells , 2004, Clinical & Experimental Metastasis.