Synergistic Effect of Focal Adhesion Kinase Overexpression and Hepatocyte Growth Factor Stimulation on Cell Transformation

Abstract

Although an elevated level of focal adhesion kinase (FAK) has been observed in a variety of invasive human tumors, forced expression of FAK alone in cultured cells does not cause them to exhibit transformed phenotypes. Therefore, the role of FAK in oncogenic transformation remains unclear. In this study, we have demonstrated that FAK overexpression in Madin-Darby canine kidney epithelial cells rendered them susceptible to transformation by hepatocyte growth factor (HGF). Using various FAK mutants, we found that the simultaneous bindings of Src and p130 cas were required for FAK to potentiate cell transformation. Expression of FAK-related nonkinase, kinase-deficient Src, or the Src homology 3 domain of p130 cas , which respectively serve as dominant negative versions of FAK, Src, and p130 cas , apparently reversed the transformed phenotypes of FAK-overexpressed cells upon HGF stimulation. Moreover, FAK overexpression was able to enhance HGF-elicited signals, leading to sustained activation of ERK, JNK, and AKT, which could be prevented by the expression of the Src homology 3 domain of p130 cas . Taken together, our results indicate that the synergistic effect of FAK overexpression and HGF stimulation leads to cell transformation and implicate a critical role of p130 cas in this process. Although an elevated level of focal adhesion kinase (FAK) has been observed in a variety of invasive human tumors, forced expression of FAK alone in cultured cells does not cause them to exhibit transformed phenotypes. Therefore, the role of FAK in oncogenic transformation remains unclear. In this study, we have demonstrated that FAK overexpression in Madin-Darby canine kidney epithelial cells rendered them susceptible to transformation by hepatocyte growth factor (HGF). Using various FAK mutants, we found that the simultaneous bindings of Src and p130 cas were required for FAK to potentiate cell transformation. Expression of FAK-related nonkinase, kinase-deficient Src, or the Src homology 3 domain of p130 cas , which respectively serve as dominant negative versions of FAK, Src, and p130 cas , apparently reversed the transformed phenotypes of FAK-overexpressed cells upon HGF stimulation. Moreover, FAK overexpression was able to enhance HGF-elicited signals, leading to sustained activation of ERK, JNK, and AKT, which could be prevented by the expression of the Src homology 3 domain of p130 cas . Taken together, our results indicate that the synergistic effect of FAK overexpression and HGF stimulation leads to cell transformation and implicate a critical role of p130 cas in this process. focal adhesion kinase hepatocyte growth factor phosphatidylinositol 3-kinase c-Jun NH2-terminal kinase extracellular signal-regulated kinase Madin-Darby canine kidney hemagglutinin FAK-related nonkinase wild type kinase-deficient Src homology 3 the SH3 domain of p130 cas Focal adhesion kinase (FAK),1 a 125-kDa cytoplasmic protein-tyrosine kinase localized in focal contacts, has been implicated to play a crucial role in the control of integrin-mediated cellular functions including cell spreading (1Richardson A. Malik R.K. Hildebrand J.D. Parsons J.T. Mol. Cell. Biol. 1997; 17: 6906-6914Crossref PubMed Scopus (290) Google Scholar, 2Richardson A. Parsons J.T. Nature. 1996; 380: 538-540Crossref PubMed Scopus (454) Google Scholar), cell migration (3Cary L.A. Chang J.F. Guan J.-L. J. Cell Sci. 1996; 109: 1787-1794Crossref PubMed Google Scholar,4Ilic D. Furuta Y. Kanazawa S. Takeda N. Sobue K. Nakatsuji N. Nomura S. Fujimoto J. Okada M. Yamamoto T. Aizawa S. Nature. 1995; 377: 539-544Crossref PubMed Scopus (1602) Google Scholar), cell cycle progression (5Oktay M. Wary K.K. Dans M. Rirge R.B. Giancotti F.G. J. Cell Biol. 1999; 145: 1461-1469Crossref PubMed Scopus (250) Google Scholar, 6Zhao J.-H. Reiske H. Guan J.-L. J. Cell Biol. 1998; 143: 1997-2008Crossref PubMed Scopus (304) Google Scholar), and cell survival (7Almeida E.A.C. Ilic D. Han Q. Hauck C.R. Jin F. Kawakatsu H. Schlaepfer D.D. Damsky C.H. J. Cell Biol. 2000; 149: 741-754Crossref PubMed Scopus (336) Google Scholar, 8Chan P.-C. Lai J.-F. Cheng C.-H. Tang M.-J. Chiu C.-C. Chen H.-C. J. Biol. Chem. 1999; 274: 26901-26906Abstract Full Text Full Text PDF PubMed Scopus (101) Google Scholar, 9Frisch S.M. Vuori K. Ruoslahti E. Chan-Hui P.-Y. J. Cell Biol. 1996; 134: 793-799Crossref PubMed Scopus (1008) Google Scholar, 10Ilic D. Almeida E.A.C. Schlaepfer D.D. Dazin P. Aizawa S. Damsky C.H. J. Cell Biol. 1998; 143: 547-560Crossref PubMed Scopus (439) Google Scholar). The ability of FAK to transduce signals to the downstream depends on its ability to interact with several intracellular signaling molecules, including Src family kinases (11Cobb B.S. Schaller M.D. Leu T.-H. Parsons J.T. Mol. Cell. Biol. 1994; 14: 147-155Crossref PubMed Scopus (496) Google Scholar, 12Xing Z. Chen H.-C. Nowlen J.K. Taylor S. Shalloway D. Guan J.-L. Mol. Biol. Cell. 1994; 5: 413-421Crossref PubMed Scopus (292) Google Scholar), phosphatidylinositol 3-kinase (PI3K) (13Chen H.-C. Guan J.-L. Proc. Natl. Acad. Sci. U. S. A. 1994; 91: 10148-10152Crossref PubMed Scopus (486) Google Scholar, 14Chen H.-C. Appeddu P.A. Isoda H. Guan J.-L. J. Biol. Chem. 1996; 271: 26329-26334Abstract Full Text Full Text PDF PubMed Scopus (474) Google Scholar), phospholipase C-γ1 (15Zhang X. Chattopadhyay A., Ji, Q.-S. Owen J.D. Ruest P.J. Carpenter G. Hanks S.T. Proc. Natl. Acad. Sci. U. S. A. 1999; 96: 9021-9026Crossref PubMed Scopus (160) Google Scholar), Grb2 (16Schlaepfer D.D. Hanks S.K. Hunter T. van der Geer P. Nature. 1994; 372: 786-791Crossref PubMed Scopus (1462) Google Scholar, 17Schlaepfer D.D. Hunter T. Mol. Cell. Biol. 1996; 16: 5623-5633Crossref PubMed Scopus (401) Google Scholar), and p130 cas (18Polte T.R. Hanks S.K. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 10678-10682Crossref PubMed Scopus (392) Google Scholar). We have previously shown that the simultaneous bindings of PI3K and p130 cas are required for FAK to promote cell migration (19Reiske H.R. Kao S.-C. Cary L.A. Guan J.-L. Lai J.-F. Chen H.-C. J. Biol. Chem. 1999; 274: 12361-12366Abstract Full Text Full Text PDF PubMed Scopus (235) Google Scholar) and survival (8Chan P.-C. Lai J.-F. Cheng C.-H. Tang M.-J. Chiu C.-C. Chen H.-C. J. Biol. Chem. 1999; 274: 26901-26906Abstract Full Text Full Text PDF PubMed Scopus (101) Google Scholar). Almeida et al. (7Almeida E.A.C. Ilic D. Han Q. Hauck C.R. Jin F. Kawakatsu H. Schlaepfer D.D. Damsky C.H. J. Cell Biol. 2000; 149: 741-754Crossref PubMed Scopus (336) Google Scholar) showed that the FAK-p130 cas complex transduces matrix survival signals via c-Jun NH2-terminal kinase (JNK). This FAK-p130 cas -JNK signaling pathway was also shown to be required for FAK to promote cell cycle progression (5Oktay M. Wary K.K. Dans M. Rirge R.B. Giancotti F.G. J. Cell Biol. 1999; 145: 1461-1469Crossref PubMed Scopus (250) Google Scholar). The FAK-Grb2 complex has been proposed to trigger downstream signaling pathways, leading to activation of extracellular signal-regulated kinases (ERKs) (17Schlaepfer D.D. Hunter T. Mol. Cell. Biol. 1996; 16: 5623-5633Crossref PubMed Scopus (401) Google Scholar, 20Schlaepfer D.D. Hunter T. J. Biol. Chem. 1997; 272: 13189-13195Abstract Full Text Full Text PDF PubMed Scopus (351) Google Scholar), which has recently been shown to contribute partially to FAK-promoted cell migration (21Lai J.-F. Kao S.-C. Jiang S.-T. Tang M.-J. Chan P.-C. Chen H.-C. J. Biol. Chem. 2000; 275: 7474-7480Abstract Full Text Full Text PDF PubMed Scopus (70) Google Scholar). Appropriate cell behavior requires coordinate signals from both cell adhesion and growth factors. Evidence has suggested that FAK may be a point of convergence of integrin and growth factor signaling pathways. In addition to cell adhesion, the tyrosine phosphorylation of FAK is also stimulated by growth factors (22Chen H.-C. Chan P.-C. Tang M.-J. Cheng C.-H. Chang T.-J. J. Biol. Chem. 1998; 273: 25777-25782Abstract Full Text Full Text PDF PubMed Scopus (82) Google Scholar, 23Hatai M. Hashi H. Mogi A. Soga H. Yokota J. Yaoi Y. FEBS Lett. 1994; 350: 113-116Crossref PubMed Scopus (42) Google Scholar, 24Matsumoto K. Matsumoto K. Nakamura T. Kramer R.H. J. Biol. Chem. 1994; 269: 31807-31813Abstract Full Text PDF PubMed Google Scholar, 25Rankin S. Rozengurt E. J. Biol. Chem. 1994; 269: 704-710Abstract Full Text PDF PubMed Google Scholar) as a result of FAK association with growth factor receptor (26Sieg D.J. Hauck C.R. Ilic D. Klingbell C.K. Schaefer E. Damsky C.H. Schlaepfer D.D. Nat. Cell Biol. 2000; 2: 249-256Crossref PubMed Scopus (1080) Google Scholar) and/or Src activation by growth factor receptor (22Chen H.-C. Chan P.-C. Tang M.-J. Cheng C.-H. Chang T.-J. J. Biol. Chem. 1998; 273: 25777-25782Abstract Full Text Full Text PDF PubMed Scopus (82) Google Scholar). The tyrosine phosphorylation of FAK stimulated by growth factors enhances its association with effectors, leading to amplification of downstream signals (22Chen H.-C. Chan P.-C. Tang M.-J. Cheng C.-H. Chang T.-J. J. Biol. Chem. 1998; 273: 25777-25782Abstract Full Text Full Text PDF PubMed Scopus (82) Google Scholar, 27Chen H.-C. Guan J.-L. J. Biol. Chem. 1994; 269: 31229-31233Abstract Full Text PDF PubMed Google Scholar). More recently, FAK was shown to integrate signals from growth factor receptors and integrins to facilitate cell migration (21Lai J.-F. Kao S.-C. Jiang S.-T. Tang M.-J. Chan P.-C. Chen H.-C. J. Biol. Chem. 2000; 275: 7474-7480Abstract Full Text Full Text PDF PubMed Scopus (70) Google Scholar, 26Sieg D.J. Hauck C.R. Ilic D. Klingbell C.K. Schaefer E. Damsky C.H. Schlaepfer D.D. Nat. Cell Biol. 2000; 2: 249-256Crossref PubMed Scopus (1080) Google Scholar). Hepatocyte growth factor (HGF), also known as scatter factor, is a growth factor that and in various cell Matsumoto K. Nakamura T. Cell. Full Text PDF PubMed Scopus Google Scholar, Schaller G. Cell. Full Text PDF PubMed Scopus Google Scholar, A. A. Mol. Cell. Biol. 1995; PubMed Google Scholar, J. Cell Biol. 1995; PubMed Scopus Google Scholar). The of HGF are activation of its receptor by the Chan PubMed Scopus Google Scholar, E. P. M. Mol. Cell. Biol. PubMed Scopus Google Scholar). activation of signaling has been implicated in the of a of human and has been shown to invasive and to cells Matsumoto K. Kramer R.H. J. 1997; PubMed Scopus Google Scholar, M. A. H. E. S. S. S. M. 1995; Google Scholar, M. T. A. A. G. S. G. 2000; PubMed Scopus (292) Google Scholar, S. A. Z. S. Proc. Natl. Acad. Sci. U. S. A. 1997; PubMed Scopus Google Scholar, M. S. J. Mol. 1996; PubMed Scopus Google Scholar). an level of FAK has been found in a variety of invasive human and has been implicated to play a role in progression to an invasive M. 2000; Scopus Google Scholar, 1995; Google Scholar, PubMed Scopus Google Scholar). We have previously shown that FAK overexpression in Madin-Darby canine kidney cells enhances migration in to HGF stimulation (21Lai J.-F. Kao S.-C. Jiang S.-T. Tang M.-J. Chan P.-C. Chen H.-C. J. Biol. Chem. 2000; 275: 7474-7480Abstract Full Text Full Text PDF PubMed Scopus (70) Google Scholar), that the FAK overexpression and HGF stimulation cell In this study, we this leads to cell transformation. human HGF was from and were from and were from was from The for was from The was previously Z. Chen H.-C. Nowlen J.K. Taylor S. Shalloway D. Guan J.-L. Mol. Biol. Cell. 1994; 5: 413-421Crossref PubMed Scopus (292) Google Scholar). The and were from The was from The was from The was from The cas and were from The and were from The expression FAK-related nonkinase was previously (21Lai J.-F. Kao S.-C. Jiang S.-T. Tang M.-J. Chan P.-C. Chen H.-C. J. Biol. Chem. 2000; 275: 7474-7480Abstract Full Text Full Text PDF PubMed Scopus (70) Google Scholar). The kinase-deficient Src was by Shalloway cells wild type FAK, FAK and or have been previously (8Chan P.-C. Lai J.-F. Cheng C.-H. Tang M.-J. Chiu C.-C. Chen H.-C. J. Biol. Chem. 1999; 274: 26901-26906Abstract Full Text Full Text PDF PubMed Scopus (101) Google Scholar, J.-F. Kao S.-C. Jiang S.-T. Tang M.-J. Chan P.-C. Chen H.-C. J. Biol. Chem. 2000; 275: 7474-7480Abstract Full Text Full Text PDF PubMed Scopus (70) Google Scholar) and were in with and cells FAK and the SH3 domain p130 cas have been previously (8Chan P.-C. Lai J.-F. Cheng C.-H. Tang M.-J. Chiu C.-C. Chen H.-C. J. Biol. Chem. 1999; 274: 26901-26906Abstract Full Text Full Text PDF PubMed Scopus (101) Google Scholar) and were in growth and cells FAK and cells that FAK were on and with of and of of the were in growth and and for FAK and expression by with cells FAK and Src, cells that FAK were with of and of were in growth and and by in Src cells were in of and with or HGF and a of and in a of and with or HGF was to the was in the of was the cells were with HGF in for and by cells in of were to an of the that been with of in of with was to the cells that and with were and a was in were in and and The were for to and the were the of were with 3 of various or of or for were on were with The were with for 3 in to and to and was with the for Src were with and in In kinase were in of kinase of and of for were by the addition of and were by the PI3K with FAK in FAK were with from cell and to PI3K as previously (13Chen H.-C. Guan J.-L. Proc. Natl. Acad. Sci. U. S. A. 1994; 91: 10148-10152Crossref PubMed Scopus (486) Google Scholar). expression of FAK and to HGF to cell cell FAK were and to the and the in the or of The expression of FAK in cells to a in FAK and the phosphorylation of FAK was upon HGF stimulation The phosphorylation of was to a level in control cells and FAK-overexpressed cells upon HGF stimulation of of canine in of was to the expression of We found that the of was not by FAK overexpression in the or of HGF not Therefore, is that FAK overexpression enhances the activation of upon HGF stimulation. The has been to cell a of cell transformation. shown in FAK-overexpressed cells to in the of that elevated expression of FAK by is not to an to HGF stimulation alone was not to the growth of control cells in HGF stimulation FAK-overexpressed cells to in and cell that the synergistic effect of FAK overexpression and HGF stimulation leads to cell is of transformed FAK-overexpressed cells invasive in to HGF cell FAK were with HGF for and to This the of cells to a and of cell process. In the of HGF cells not exhibit invasive FAK was Although HGF has been shown to the of various cell K. Matsumoto K. Nakamura T. Kramer R.H. J. Biol. Chem. 1994; 269: 31807-31813Abstract Full Text PDF PubMed Google Scholar, M. S. Mol. Cell. Biol. 1996; 16: PubMed Scopus Google Scholar, T. Matsumoto K. A. Y. Nakamura T. 1997; Google Scholar), to promote control cells to our HGF stimulation the invasive of cells FAK In addition to the FAK overexpression and HGF stimulation was in Chang FAK overexpression in Chang cells them susceptible to transformation upon HGF from the in transformation not the effect of FAK overexpression on cell a dominant negative of FAK, was in cells that FAK Although has been to be in cell X. Cell 1998; Google Scholar), is not the for a growth not cell not result showed that the expression of FAK-overexpressed cells to in and upon HGF stimulation the role of FAK overexpression in cell transformation. the signals downstream of FAK required for cell cell FAK including and in to Src, p130 cas , and were We demonstrated that the FAK in cells was to the in the FAK signaling 3 cells were to the and the We found that the cells FAK or to in and upon HGF stimulation that simultaneous bindings of Src and p130 cas are required for FAK to potentiate cell transformation. The of cells FAK to in and to upon HGF stimulation were to of cells FAK, that the PI3K may be for FAK to cell transformation upon HGF stimulation. cells the FAK invasive HGF in of that of cells FAK, that the of FAK and Grb2 may contribute to cell growth and demonstrated in the cas signaling pathway is to be critical for FAK to potentiate the cell transformation. the of Src in this Src, which functions as a dominant negative of Src, was in cells that FAK Although the expression of Src not apparently the of Src the Src and the tyrosine phosphorylation of FAK stimulated by HGF an role for Src in FAK as previously suggested by our (22Chen H.-C. Chan P.-C. Tang M.-J. Cheng C.-H. Chang T.-J. J. Biol. Chem. 1998; 273: 25777-25782Abstract Full Text Full Text PDF PubMed Scopus (82) Google Scholar). Moreover, the expression of Src the of FAK-overexpressed cells to in and upon HGF stimulation by the Src family kinase was found to the transformed phenotypes of FAK-overexpressed cells upon HGF stimulation the of p130 cas in this transformation the SH3 domain of p130 cas was in cells that FAK We have previously demonstrated that the SH3 domain of p130 cas with p130 cas for FAK (8Chan P.-C. Lai J.-F. Cheng C.-H. Tang M.-J. Chiu C.-C. Chen H.-C. J. Biol. Chem. 1999; 274: 26901-26906Abstract Full Text Full Text PDF PubMed Scopus (101) Google Scholar, H.R. Kao S.-C. Cary L.A. Guan J.-L. Lai J.-F. Chen H.-C. J. Biol. Chem. 1999; 274: 12361-12366Abstract Full Text Full Text PDF PubMed Scopus (235) Google Scholar). In this study, we showed that the expression of the SH3 domain of p130 cas in FAK-overexpressed cells to in and to upon HGF stimulation results the of the cas signaling in cell of HGF-elicited by FAK control cells and cells FAK or both FAK and were for and with HGF for or of cell was by with as result of is cells FAK were to the and the in the of HGF and an or was in both The was as a are from and were on the level of cells in the of HGF and effect of the SH3 domain of p130 cas on FAK of cell transformation. cell from control cells and cells FAK, or both FAK and were by with cells as were to the and the in the of are from from for and were on the level of cells FAK the in FAK of cell control cells and cell FAK with or the expression of the SH3 domain of p130 cas were and to HGF for a or of The activation of intracellular signaling that have been to be in HGF signaling was by with HGF stimulation to sustained activation of ERK, JNK, and in FAK-overexpressed cells activation of in control Although and phospholipase were to be by HGF M. A. P. Nature. 1998; PubMed Scopus Google Scholar, A. Z. F. P. S. A. G. Cell. 1994; Full Text PDF PubMed Scopus Google Scholar), phosphorylation were not in our not The expression of the SH3 domain of p130 cas was of the sustained activation of ERK, JNK, and by both FAK overexpression and HGF stimulation. Taken together, we the that signaling in this may in FAK of cell the sustained of ERK, JNK, and are to partially for the of transformed phenotypes. we found that the kinase signal-regulated kinase transformation of FAK-overexpressed cells the of the pathway in this transformation In the could the cell transformation that is required for cell transformation by the FAK overexpression and HGF stimulation. FAK has been implicated to play a critical role in integrin-mediated cellular functions including cell cell cycle and cell is that an level of FAK may contribute to a in the role of FAK in has not been In this study, we have an in to the effect of expression of FAK on cell transformation. The results in this that a FAK overexpression and HGF stimulation leads to cell from in cell growth and FAK overexpression HGF stimulation alone is of Therefore, our results that an elevated expression of FAK may cells to HGF leading to amplification of intracellular signals for the control of cellular is that FAK may play in crucial by its to an growth and to promote of cells upon growth factor stimulation. has previously been shown that the expression of the of FAK by to the cells upon and to an growth in and in S.M. Vuori K. Ruoslahti E. Chan-Hui P.-Y. J. Cell Biol. 1996; 134: 793-799Crossref PubMed Scopus (1008) Google Scholar). the and phosphorylation of FAK be by HGF stimulation in a of integrin-mediated cell adhesion (22Chen H.-C. Chan P.-C. Tang M.-J. Cheng C.-H. Chang T.-J. J. Biol. Chem. 1998; 273: 25777-25782Abstract Full Text Full Text PDF PubMed Scopus (82) Google Scholar), is that signals by the FAK overexpression and HGF stimulation may be to by integrins and growth factor receptors in signaling pathways, is that are for integrin signals to growth factor signals and integrins various growth factor receptors including to to the of tyrosine phosphorylation cell adhesion Proc. Natl. Acad. Sci. U. S. A. 1996; PubMed Scopus Google Scholar). FAK is known to be by cell adhesion, was to be in this process. FAK overexpression not to enhance the phosphorylation of in the or of HGF stimulation that the transformed cell observed in this is not the result of an activation of by FAK In FAK has been demonstrated not to be in cell activation of or growth factor receptor Proc. Natl. Acad. Sci. U. S. A. 1996; PubMed Scopus Google Scholar, S. E. M. E. M. J. A. Schaefer E. P. G. P. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). our results that FAK is to serve as a to potentiate activation of signaling pathways. in human kidney cells (22Chen H.-C. Chan P.-C. Tang M.-J. Cheng C.-H. Chang T.-J. J. Biol. Chem. 1998; 273: 25777-25782Abstract Full Text Full Text PDF PubMed Scopus (82) Google Scholar), the expression of Src or the addition of a Src FAK phosphorylation in cells We that Src in is to FAK of integrin an signaling to potentiate HGF-elicited HGF may cause the tyrosine phosphorylation of integrins as A. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar), which FAK to FAK has been shown to be by integrin M. Cheng H.-C. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). In this study, we have found that FAK in Src or p130 cas to cells with HGF that simultaneous of both Src and p130 cas are required for FAK to its In the that the expression of the SH3 domain of p130 cas in of transformed phenotypes of FAK-overexpressed cells the of p130 cas in this transformation p130 cas has previously been shown to be for FAK to promote cell survival (7Almeida E.A.C. Ilic D. Han Q. Hauck C.R. Jin F. Kawakatsu H. Schlaepfer D.D. Damsky C.H. J. Cell Biol. 2000; 149: 741-754Crossref PubMed Scopus (336) Google Scholar, 8Chan P.-C. Lai J.-F. Cheng C.-H. Tang M.-J. Chiu C.-C. Chen H.-C. J. Biol. Chem. 1999; 274: 26901-26906Abstract Full Text Full Text PDF PubMed Scopus (101) Google Scholar) and migration (21Lai J.-F. Kao S.-C. Jiang S.-T. Tang M.-J. Chan P.-C. Chen H.-C. J. Biol. Chem. 2000; 275: 7474-7480Abstract Full Text Full Text PDF PubMed Scopus (70) Google L.A. Han T.R. Hanks S.K. Guan J.-L. J. Cell Biol. 1998; PubMed Scopus Google Scholar). is that p130 cas may contribute to cell transformation by its in cell survival and both of which are to growth and In p130 cas was as a in cells transformed by Parsons J.T. Mol. Cell. Biol. PubMed Scopus Google Scholar) or H. Proc. Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar). phosphorylation and association with has been shown to be critical for its J. Vuori K. J. Cell Biol. 1998; PubMed Scopus Google Scholar). Src may contribute to cell transformation by of p130 cas , its in FAK of transformation may be the result of its in p130 cas . We have previously shown that FAK the to cells from (8Chan P.-C. Lai J.-F. Cheng C.-H. Tang M.-J. Chiu C.-C. Chen H.-C. J. Biol. Chem. 1999; 274: 26901-26906Abstract Full Text Full Text PDF PubMed Scopus (101) Google Scholar) and to promote cell migration on matrix (21Lai J.-F. Kao S.-C. Jiang S.-T. Tang M.-J. Chan P.-C. Chen H.-C. J. Biol. Chem. 2000; 275: 7474-7480Abstract Full Text Full Text PDF PubMed Scopus (70) Google Scholar). In this study, we found that the FAK to was able to a transformed to cells upon HGF stimulation and that the of PI3K is for FAK to its upon HGF stimulation. our results not the of PI3K in this transformation is known that HGF PI3K in cells A. K. J. J. Biol. Chem. 1998; 273: Full Text Full Text PDF PubMed Scopus Google Scholar, M. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google is that the of PI3K activation by HGF stimulation may have its for cell of the of PI3K by with we found that the PI3K FAK-overexpressed cells from in and to upon HGF stimulation not the downstream signals for cell transformation by the synergistic effect of FAK overexpression and HGF the HGF-elicited activation of intracellular signaling was FAK-overexpressed cells and control the in this study, the of ERK, JNK, and activation by HGF stimulation was by FAK that the sustained activation of signaling may partially for cell transformation in this In of the effect of the SH3 domain of p130 cas on the sustained activation of ERK, JNK, and AKT, is that the signals downstream of p130 cas may to In the signals by p130 cas have been shown to Y. S. Mol. Cell. Biol. 1999; PubMed Scopus (42) Google Scholar) and (5Oktay M. Wary K.K. Dans M. Rirge R.B. Giancotti F.G. J. Cell Biol. 1999; 145: 1461-1469Crossref PubMed Scopus (250) Google Scholar, E.A.C. Ilic D. Han Q. Hauck C.R. Jin F. Kawakatsu H. Schlaepfer D.D. Damsky C.H. J. Cell Biol. 2000; 149: 741-754Crossref PubMed Scopus (336) Google the for p130 cas to in this remains to be The HGF receptor is the by the Chan PubMed Scopus Google Scholar, E. P. M. Mol. Cell. Biol. PubMed Scopus Google Scholar). activation of the pathway has been implicated in the of a of human and has been shown to invasive and to cells Matsumoto K. Kramer R.H. J. 1997; PubMed Scopus Google Scholar, M. A. H. E. S. S. S. M. 1995; Google Scholar, M. T. A. A. G. S. G. 2000; PubMed Scopus (292) Google Scholar, S. A. Z. S. Proc. Natl. Acad. Sci. U. S. A. 1997; PubMed Scopus Google Scholar, M. S. J. Mol. 1996; PubMed Scopus Google Scholar). an elevated level of FAK has been implicated to play a role in progression to an invasive M. 2000; Scopus Google Scholar, 1995; Google Scholar, PubMed Scopus Google Scholar). In this study, we that an level of FAK may cells susceptible to transformation by HGF a that does not cell a signaling and FAK In addition to several growth factors have been shown to FAK (22Chen H.-C. Chan P.-C. Tang M.-J. Cheng C.-H. Chang T.-J. J. Biol. Chem. 1998; 273: 25777-25782Abstract Full Text Full Text PDF PubMed Scopus (82) Google Scholar, 23Hatai M. Hashi H. Mogi A. Soga H. Yokota J. Yaoi Y. FEBS Lett. 1994; 350: 113-116Crossref PubMed Scopus (42) Google Scholar, 24Matsumoto K. Matsumoto K. Nakamura T. Kramer R.H. J. Biol. Chem. 1994; 269: 31807-31813Abstract Full Text PDF PubMed Google Scholar, 25Rankin S. Rozengurt E. J. Biol. Chem. 1994; 269: 704-710Abstract Full Text PDF PubMed Google Scholar). growth factor also a transformed in cells FAK, to a not Therefore, our results implicate that expression of FAK may growth factors in and the of FAK expression level as a for in We Shalloway for