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FAK signaling is critical for ErbB-2/ErbB-3 receptor cooperation for oncogenic transformation and invasion.

Benlimame N, He Q, Jie S, Xiao D, Xu YJ, Loignon M, Schlaepfer DD, Alaoui-Jamali MA - J. Cell Biol. (2005)

Bottom Line: The overexpression of members of the ErbB tyrosine kinase receptor family has been associated with cancer progression.This colocalization requires intact FAK.In summary, distinct FAK signaling has an essential function in ErbB-induced oncogenesis and invasiveness.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Lady Davis Institute of the Sir Mortimer B. Davis Jewish General Hospital, McGill University, Montreal, Quebec H3T 1E2, Canada.

ABSTRACT
The overexpression of members of the ErbB tyrosine kinase receptor family has been associated with cancer progression. We demonstrate that focal adhesion kinase (FAK) is essential for oncogenic transformation and cell invasion that is induced by ErbB-2 and -3 receptor signaling. ErbB-2/3 overexpression in FAK-deficient cells fails to promote cell transformation and rescue chemotaxis deficiency. Restoration of FAK rescues both oncogenic transformation and invasion that is induced by ErbB-2/3 in vitro and in vivo. In contrast, the inhibition of FAK in FAK-proficient invasive cancer cells prevented cell invasion and metastasis formation. The activation of ErbB-2/3 regulates FAK phosphorylation at Tyr-397, -861, and -925. ErbB-induced oncogenic transformation correlates with the ability of FAK to restore ErbB-2/3-induced mitogen-activated protein kinase (MAPK) activation; the inhibition of MAPK prevented oncogenic transformation. In contrast, the inhibition of Src but not MAPK prevented ErbB-FAK-induced chemotaxis. In migratory cells, activated ErbB-2/3 receptors colocalize with activated FAK at cell protrusions. This colocalization requires intact FAK. In summary, distinct FAK signaling has an essential function in ErbB-induced oncogenesis and invasiveness.

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FAK is essential for ErbB-induced oncogenic transformation and chemotaxis. (A) Western blot analysis to show FAK expression status in FAK−/− cells expressing various ErbB receptor combinations and their matched cells in which wild-type FAK was reconstituted by stable transfection, as indicated in Materials and methods. FAK+/+–ErbB-2/3 cells are included as a control. (B) Oncogenic property of FAK−/−- and FAK+/+-expressing ErbB-2/3 receptors. Cells were cultured in medium containing soft agarose, and colony formation was determined 4 wk later by counting the number of cell foci of >20 μm in diameter (Fig. S2 A, available at http://www.jcb.org/cgi/content/full/jcb.200504124/DC1). Each bar on the graph represents the mean number of colonies per well from three independent experiments ± SD. (C and D) ErbB-induced cell invasion in FAK+/+ (C), FAK−/−, and FAK−/− cells in which FAK was restored (D). FAK+/+, FAK−/−, and FAK−/−-reconstituted cells expressing ErbB receptors were cultured in the upper chamber, whereas EGF or HRG was used as a chemoattractant in the lower chamber. Each bar of the graphs represents the mean ± SD (error bars) of invading cells from three independent experiments. C, control.
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fig2: FAK is essential for ErbB-induced oncogenic transformation and chemotaxis. (A) Western blot analysis to show FAK expression status in FAK−/− cells expressing various ErbB receptor combinations and their matched cells in which wild-type FAK was reconstituted by stable transfection, as indicated in Materials and methods. FAK+/+–ErbB-2/3 cells are included as a control. (B) Oncogenic property of FAK−/−- and FAK+/+-expressing ErbB-2/3 receptors. Cells were cultured in medium containing soft agarose, and colony formation was determined 4 wk later by counting the number of cell foci of >20 μm in diameter (Fig. S2 A, available at http://www.jcb.org/cgi/content/full/jcb.200504124/DC1). Each bar on the graph represents the mean number of colonies per well from three independent experiments ± SD. (C and D) ErbB-induced cell invasion in FAK+/+ (C), FAK−/−, and FAK−/− cells in which FAK was restored (D). FAK+/+, FAK−/−, and FAK−/−-reconstituted cells expressing ErbB receptors were cultured in the upper chamber, whereas EGF or HRG was used as a chemoattractant in the lower chamber. Each bar of the graphs represents the mean ± SD (error bars) of invading cells from three independent experiments. C, control.

Mentions: Parental FAK−/− and FAK+/+ cells exhibit no apparent morphological changes that reflect cell transformation, and they lack the ability to grow on soft agar and form tumors in immunocompromised mice (see Fig. 3). Neither control FAK+/+ nor FAK−/− cells expressing empty retroviral particles that were used to express ErbB receptors formed colonies in soft agar. In contrast, FAK+/+-2 and -2/3 cells, but not FAK+/+-3 or any FAK−/−–ErbB-expressing cells, were able to grow on soft agar and form large foci; the coexpression of ErbB-2 with -3 resulted in strong oncogenic transformation compared with ErbB-2 alone (Fig. 2 B and Fig. S2 A, available at http://www.jcb.org/cgi/content/full/jcb.200504124/DC1).


FAK signaling is critical for ErbB-2/ErbB-3 receptor cooperation for oncogenic transformation and invasion.

Benlimame N, He Q, Jie S, Xiao D, Xu YJ, Loignon M, Schlaepfer DD, Alaoui-Jamali MA - J. Cell Biol. (2005)

FAK is essential for ErbB-induced oncogenic transformation and chemotaxis. (A) Western blot analysis to show FAK expression status in FAK−/− cells expressing various ErbB receptor combinations and their matched cells in which wild-type FAK was reconstituted by stable transfection, as indicated in Materials and methods. FAK+/+–ErbB-2/3 cells are included as a control. (B) Oncogenic property of FAK−/−- and FAK+/+-expressing ErbB-2/3 receptors. Cells were cultured in medium containing soft agarose, and colony formation was determined 4 wk later by counting the number of cell foci of >20 μm in diameter (Fig. S2 A, available at http://www.jcb.org/cgi/content/full/jcb.200504124/DC1). Each bar on the graph represents the mean number of colonies per well from three independent experiments ± SD. (C and D) ErbB-induced cell invasion in FAK+/+ (C), FAK−/−, and FAK−/− cells in which FAK was restored (D). FAK+/+, FAK−/−, and FAK−/−-reconstituted cells expressing ErbB receptors were cultured in the upper chamber, whereas EGF or HRG was used as a chemoattractant in the lower chamber. Each bar of the graphs represents the mean ± SD (error bars) of invading cells from three independent experiments. C, control.
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Related In: Results  -  Collection

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fig2: FAK is essential for ErbB-induced oncogenic transformation and chemotaxis. (A) Western blot analysis to show FAK expression status in FAK−/− cells expressing various ErbB receptor combinations and their matched cells in which wild-type FAK was reconstituted by stable transfection, as indicated in Materials and methods. FAK+/+–ErbB-2/3 cells are included as a control. (B) Oncogenic property of FAK−/−- and FAK+/+-expressing ErbB-2/3 receptors. Cells were cultured in medium containing soft agarose, and colony formation was determined 4 wk later by counting the number of cell foci of >20 μm in diameter (Fig. S2 A, available at http://www.jcb.org/cgi/content/full/jcb.200504124/DC1). Each bar on the graph represents the mean number of colonies per well from three independent experiments ± SD. (C and D) ErbB-induced cell invasion in FAK+/+ (C), FAK−/−, and FAK−/− cells in which FAK was restored (D). FAK+/+, FAK−/−, and FAK−/−-reconstituted cells expressing ErbB receptors were cultured in the upper chamber, whereas EGF or HRG was used as a chemoattractant in the lower chamber. Each bar of the graphs represents the mean ± SD (error bars) of invading cells from three independent experiments. C, control.
Mentions: Parental FAK−/− and FAK+/+ cells exhibit no apparent morphological changes that reflect cell transformation, and they lack the ability to grow on soft agar and form tumors in immunocompromised mice (see Fig. 3). Neither control FAK+/+ nor FAK−/− cells expressing empty retroviral particles that were used to express ErbB receptors formed colonies in soft agar. In contrast, FAK+/+-2 and -2/3 cells, but not FAK+/+-3 or any FAK−/−–ErbB-expressing cells, were able to grow on soft agar and form large foci; the coexpression of ErbB-2 with -3 resulted in strong oncogenic transformation compared with ErbB-2 alone (Fig. 2 B and Fig. S2 A, available at http://www.jcb.org/cgi/content/full/jcb.200504124/DC1).

Bottom Line: The overexpression of members of the ErbB tyrosine kinase receptor family has been associated with cancer progression.This colocalization requires intact FAK.In summary, distinct FAK signaling has an essential function in ErbB-induced oncogenesis and invasiveness.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Lady Davis Institute of the Sir Mortimer B. Davis Jewish General Hospital, McGill University, Montreal, Quebec H3T 1E2, Canada.

ABSTRACT
The overexpression of members of the ErbB tyrosine kinase receptor family has been associated with cancer progression. We demonstrate that focal adhesion kinase (FAK) is essential for oncogenic transformation and cell invasion that is induced by ErbB-2 and -3 receptor signaling. ErbB-2/3 overexpression in FAK-deficient cells fails to promote cell transformation and rescue chemotaxis deficiency. Restoration of FAK rescues both oncogenic transformation and invasion that is induced by ErbB-2/3 in vitro and in vivo. In contrast, the inhibition of FAK in FAK-proficient invasive cancer cells prevented cell invasion and metastasis formation. The activation of ErbB-2/3 regulates FAK phosphorylation at Tyr-397, -861, and -925. ErbB-induced oncogenic transformation correlates with the ability of FAK to restore ErbB-2/3-induced mitogen-activated protein kinase (MAPK) activation; the inhibition of MAPK prevented oncogenic transformation. In contrast, the inhibition of Src but not MAPK prevented ErbB-FAK-induced chemotaxis. In migratory cells, activated ErbB-2/3 receptors colocalize with activated FAK at cell protrusions. This colocalization requires intact FAK. In summary, distinct FAK signaling has an essential function in ErbB-induced oncogenesis and invasiveness.

Show MeSH
Related in: MedlinePlus