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  • Original Article
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Intrinsic FAK activity and Y925 phosphorylation facilitate an angiogenic switch in tumors

Oncogene volume 25, pages 5969–5984 (2006)Cite this article

Abstract

Elevated focal adhesion kinase (FAK) expression occurs in advanced cancers, yet a signaling role for FAK in tumor progression remains undefined. Here, we suppressed FAK activity in 4T1 breast carcinoma cells resulting in reduced FAK Y925 phosphorylation, Grb2 adaptor protein binding to FAK, and signaling to mitogen-activated protein (MAP) kinase (MAPK). Loss of a FAK-Grb2-MAPK linkage did not affect 4T1 cell proliferation or survival in culture, yet FAK inhibition reduced vascular endothelial growth factor (VEGF) expression and resulted in small avascular tumors in mice. This FAK-Grb2-MAPK linkage was essential in promoting angiogenesis as reconstitution experiments using Src-transformed FAK-null fibroblasts revealed that point mutations affecting FAK catalytic activity (R454) or Y925 phosphorylation (F925) disrupted the ability of FAK to promote MAPK- and VEGF-associated tumor growth. Notably, in both FAK-inhibited 4T1 and Src-transformed FAK-null cells, constitutively activated (CA) mitogen-activated protein kinase kinase 1 (MEK1) restored VEGF production and CA-MEK1 or added VEGF rescued tumor growth and angiogenesis. These studies provide the first biological support for Y925 FAK phosphorylation and define a novel role for FAK activity in promoting a MAPK-associated angiogenic switch during tumor progression.

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Abbreviations

−/−:

null

Ad:

adenovirus

CA:

constitutively activated

CAM:

chorioallantoic membrane

Ctrl:

control

DAB:

diaminobenzidine

ERK:

extracellular signal-regulated kinase

FAK:

focal adhesion kinase

FRNK:

FAK-related non-kinase

HUVEC:

human umbilical vein endothelial cell

IVK:

in vitro kinase

MEK1:

mitogen-activated protein kinase kinase 1

PTK:

protein-tyrosine kinase

pY:

phosphotyrosine

SH2:

Src homology domain 2

v-Src:

viral Src

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Acknowledgements

We thank Bruce Torbett (Scripps) for the pVIPER lentiviral vector, Luigi Naldini (San Raffaele Telethon Institute for Gene Therapy, Milan, Italy) for the CMVDR8.74 and VSVG lentiviral packaging vectors, Charlie Surh (Scripps) for Leica cryostat usage, Christof Hauck (University of Wuerzburg, Germany) for performing the initial soft agar growth assays, Jun-Lin Guan (Cornell) for sharing results prior to publication, and we greatly appreciate the administrative assistance provided by Theresa Villalpando. S Mitra is supported by a fellowship (12FT-0122) from the California Tobacco-Related Disease Research Program and D Hanson by an NIH Training grant (T32 AI07606). This work was supported by grants from the NIH to Dusko Ilic (CA087652), Dwayne Stupack (CA107263), David Cheresh (CA50286, CA45726, CA95262, EY14174, CA78045, HL57900) and to David Schlaepfer (CA75240, CA87038, CA102310). David Schlaepfer is an Established Investigator of the American Heart Association. This is manuscript number 17598-IMM from The Scripps Research Institute.

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Author notes

  1. D A Hsia

    Present address: Graduate Program in Biomedical Sciences, University of California, Davis, USA

Authors and Affiliations

  1. Department of Immunology, The Scripps Research Institute, La Jolla, CA, USA

    S K Mitra, J E Molina, D A Hsia, D A Hanson, A Chi, S-T Lim, J A Bernard-Trifilo & D D Schlaepfer

  2. Department of Pathology, University of California San Diego, La Jolla, CA, USA

    D Mikolon, D G Stupack & D A Cheresh

  3. StemLifeLine Inc., San Carlos, CA, USA

    D Ilic

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Mitra, S., Mikolon, D., Molina, J. et al. Intrinsic FAK activity and Y925 phosphorylation facilitate an angiogenic switch in tumors. Oncogene 25, 5969–5984 (2006). https://doi.org/10.1038/sj.onc.1209588

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