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Fibroblast surface-associated FGF-2 promotes contact-dependent colorectal cancer cell migration and invasion through FGFR-SRC signaling and integrin αvβ5-mediated adhesion.

Knuchel S, Anderle P, Werfelli P, Diamantis E, Rüegg C - Oncotarget (2015)

Bottom Line: These effects depend on fibroblast cell surface-associated fibroblast growth factor (FGF) -2.Inhibition of FGF-2 or FGF receptors (FGFRs) signaling abolishes these effects.Taken together, these results demonstrate that fibroblasts induce cell-contact-dependent colorectal cancer cell migration and invasion under 2D and 3D conditions in vitro through fibroblast cell surface-associated FGF-2, FGF receptor-mediated SRC activation and αvβ5 integrin-dependent cancer cell adhesion to fibroblasts.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Faculty of Science, University of Fribourg, Fribourg, CH-1700, Switzerland.

ABSTRACT
Carcinoma-associated fibroblasts were reported to promote colorectal cancer (CRC) invasion by secreting motility factors and extracellular matrix processing enzymes. Less is known whether fibroblasts may induce CRC cancer cell motility by contact-dependent mechanisms. To address this question we characterized the interaction between fibroblasts and SW620 and HT29 colorectal cancer cells in 2D and 3D co-culture models in vitro. Here we show that fibroblasts induce contact-dependent cancer cell elongation, motility and invasiveness independently of deposited matrix or secreted factors. These effects depend on fibroblast cell surface-associated fibroblast growth factor (FGF) -2. Inhibition of FGF-2 or FGF receptors (FGFRs) signaling abolishes these effects. FGFRs activate SRC in cancer cells and inhibition or silencing of SRC in cancer cells, but not in fibroblasts, prevents fibroblasts-mediated effects. Using an RGD-based integrin antagonist and function-blocking antibodies we demonstrate that cancer cell adhesion to fibroblasts requires integrin αvβ5. Taken together, these results demonstrate that fibroblasts induce cell-contact-dependent colorectal cancer cell migration and invasion under 2D and 3D conditions in vitro through fibroblast cell surface-associated FGF-2, FGF receptor-mediated SRC activation and αvβ5 integrin-dependent cancer cell adhesion to fibroblasts. The FGF-2-FGFRs-SRC-αvβ5 integrin loop might be explored as candidate therapeutic target to block colorectal cancer invasion.

No MeSH data available.


Related in: MedlinePlus

Cancer cell migration and invasion depends of fibroblast-cell surface associated FGF-2 and FGFR signalingA. Quantification of SW620 motility cultured with fibroblasts in the absence or presence of PD-161570 and PD-173074 FGFR inhibitors for 48 h. B. Quantification of SW620 2D spheroid invasion cultured as indicated after 4 days. C. Representative images of SW620-LifeAct-mCherry 3D spheroid invasion cultured with fibroblasts-LifeAct-GFP in the absence or presence of FGFR inhibitors after 4 days. D. Western blotting analysis of FGF-2 protein in fibroblasts, cancer cells alone and in co-culture with (“(+FB)”) or without (“+FB”) cancer cell separation. E. Relative expression of FGF-2 mRNA in fibroblasts, SW620 and HT29 determined by RT-PCR. F. ELISA quantification of soluble FGF-2 in cell culture supernatants as indicated. All data are represented as mean +/− SD.
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Figure 6: Cancer cell migration and invasion depends of fibroblast-cell surface associated FGF-2 and FGFR signalingA. Quantification of SW620 motility cultured with fibroblasts in the absence or presence of PD-161570 and PD-173074 FGFR inhibitors for 48 h. B. Quantification of SW620 2D spheroid invasion cultured as indicated after 4 days. C. Representative images of SW620-LifeAct-mCherry 3D spheroid invasion cultured with fibroblasts-LifeAct-GFP in the absence or presence of FGFR inhibitors after 4 days. D. Western blotting analysis of FGF-2 protein in fibroblasts, cancer cells alone and in co-culture with (“(+FB)”) or without (“+FB”) cancer cell separation. E. Relative expression of FGF-2 mRNA in fibroblasts, SW620 and HT29 determined by RT-PCR. F. ELISA quantification of soluble FGF-2 in cell culture supernatants as indicated. All data are represented as mean +/− SD.

Mentions: In order to define the molecular mechanism involved in contact-dependent fibroblast-induced cancer cell elongation, migration and invasion, we applied pharmacological inhibitors of several known invasion-promoting kinases (i.e. PI3K/AKT, c-MET, MAPK/ERK, EGFR or FGFR) to co-cultures. Of all the tested inhibitors, only the FGFR inhibitors PD-161570 and PD-173074 prevented fibroblasts-induced cancer cell elongation (Supplementary Fig. 4A and 4B). PD-161570 and PD-173074 also inhibited cancer cell migration (Fig. 6A and Supplementary Fig. 4C) and invasion in the 2D (Fig. 6B) and 3D (Fig. 6C) assays.


Fibroblast surface-associated FGF-2 promotes contact-dependent colorectal cancer cell migration and invasion through FGFR-SRC signaling and integrin αvβ5-mediated adhesion.

Knuchel S, Anderle P, Werfelli P, Diamantis E, Rüegg C - Oncotarget (2015)

Cancer cell migration and invasion depends of fibroblast-cell surface associated FGF-2 and FGFR signalingA. Quantification of SW620 motility cultured with fibroblasts in the absence or presence of PD-161570 and PD-173074 FGFR inhibitors for 48 h. B. Quantification of SW620 2D spheroid invasion cultured as indicated after 4 days. C. Representative images of SW620-LifeAct-mCherry 3D spheroid invasion cultured with fibroblasts-LifeAct-GFP in the absence or presence of FGFR inhibitors after 4 days. D. Western blotting analysis of FGF-2 protein in fibroblasts, cancer cells alone and in co-culture with (“(+FB)”) or without (“+FB”) cancer cell separation. E. Relative expression of FGF-2 mRNA in fibroblasts, SW620 and HT29 determined by RT-PCR. F. ELISA quantification of soluble FGF-2 in cell culture supernatants as indicated. All data are represented as mean +/− SD.
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Related In: Results  -  Collection

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Figure 6: Cancer cell migration and invasion depends of fibroblast-cell surface associated FGF-2 and FGFR signalingA. Quantification of SW620 motility cultured with fibroblasts in the absence or presence of PD-161570 and PD-173074 FGFR inhibitors for 48 h. B. Quantification of SW620 2D spheroid invasion cultured as indicated after 4 days. C. Representative images of SW620-LifeAct-mCherry 3D spheroid invasion cultured with fibroblasts-LifeAct-GFP in the absence or presence of FGFR inhibitors after 4 days. D. Western blotting analysis of FGF-2 protein in fibroblasts, cancer cells alone and in co-culture with (“(+FB)”) or without (“+FB”) cancer cell separation. E. Relative expression of FGF-2 mRNA in fibroblasts, SW620 and HT29 determined by RT-PCR. F. ELISA quantification of soluble FGF-2 in cell culture supernatants as indicated. All data are represented as mean +/− SD.
Mentions: In order to define the molecular mechanism involved in contact-dependent fibroblast-induced cancer cell elongation, migration and invasion, we applied pharmacological inhibitors of several known invasion-promoting kinases (i.e. PI3K/AKT, c-MET, MAPK/ERK, EGFR or FGFR) to co-cultures. Of all the tested inhibitors, only the FGFR inhibitors PD-161570 and PD-173074 prevented fibroblasts-induced cancer cell elongation (Supplementary Fig. 4A and 4B). PD-161570 and PD-173074 also inhibited cancer cell migration (Fig. 6A and Supplementary Fig. 4C) and invasion in the 2D (Fig. 6B) and 3D (Fig. 6C) assays.

Bottom Line: These effects depend on fibroblast cell surface-associated fibroblast growth factor (FGF) -2.Inhibition of FGF-2 or FGF receptors (FGFRs) signaling abolishes these effects.Taken together, these results demonstrate that fibroblasts induce cell-contact-dependent colorectal cancer cell migration and invasion under 2D and 3D conditions in vitro through fibroblast cell surface-associated FGF-2, FGF receptor-mediated SRC activation and αvβ5 integrin-dependent cancer cell adhesion to fibroblasts.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Faculty of Science, University of Fribourg, Fribourg, CH-1700, Switzerland.

ABSTRACT
Carcinoma-associated fibroblasts were reported to promote colorectal cancer (CRC) invasion by secreting motility factors and extracellular matrix processing enzymes. Less is known whether fibroblasts may induce CRC cancer cell motility by contact-dependent mechanisms. To address this question we characterized the interaction between fibroblasts and SW620 and HT29 colorectal cancer cells in 2D and 3D co-culture models in vitro. Here we show that fibroblasts induce contact-dependent cancer cell elongation, motility and invasiveness independently of deposited matrix or secreted factors. These effects depend on fibroblast cell surface-associated fibroblast growth factor (FGF) -2. Inhibition of FGF-2 or FGF receptors (FGFRs) signaling abolishes these effects. FGFRs activate SRC in cancer cells and inhibition or silencing of SRC in cancer cells, but not in fibroblasts, prevents fibroblasts-mediated effects. Using an RGD-based integrin antagonist and function-blocking antibodies we demonstrate that cancer cell adhesion to fibroblasts requires integrin αvβ5. Taken together, these results demonstrate that fibroblasts induce cell-contact-dependent colorectal cancer cell migration and invasion under 2D and 3D conditions in vitro through fibroblast cell surface-associated FGF-2, FGF receptor-mediated SRC activation and αvβ5 integrin-dependent cancer cell adhesion to fibroblasts. The FGF-2-FGFRs-SRC-αvβ5 integrin loop might be explored as candidate therapeutic target to block colorectal cancer invasion.

No MeSH data available.


Related in: MedlinePlus