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FGF-1/-3/FGFR4 signaling in cancer-associated fibroblasts promotes tumor progression in colon cancer through Erk and MMP-7.

Bai YP, Shang K, Chen H, Ding F, Wang Z, Liang C, Xu Y, Sun MH, Li YY - Cancer Sci. (2015)

Bottom Line: The phosphorylation of FGFR4 enhanced the production of metalloproteinase (MMP)-7 and mitogen-activated protein kinase kinase (Mek)/extracellular signal-regulated kinase (Erk), which was accompanied by excessive vessel generation and cell proliferation.The administration of FGF-1/-3-neutralizing antibodies or the treatment of cells with FGFR4 siRNA or the FGFR4 inhibitor PD173074 markedly suppressed colon cancer cell proliferation and neovascularization.The inhibition of the FGF signaling pathway may be a useful strategy for the treatment of colon cancer.

View Article: PubMed Central - PubMed

Affiliation: Cancer Research Institute, Fudan University Shanghai Cancer Center, Shanghai, China.

No MeSH data available.


Related in: MedlinePlus

Cancer-associated fibroblasts (CAFs) predominantly secrete FGF1 and FGF3 and can upregulate the Mek/Erk pathway and MMP-7 through the activation of FGFR4. (a) mRNA levels of FGF-1, -2, -3, -7, EGF and HGF in NFs, PFs and CAFs were quantified through qRT-PCR. *P < 0.05 versus NFs. All experiments were repeated three times, and representative results are shown here. (b) The relative FGF1 and FGF3 mRNA levels in NFs, PFs, CAFs, and HCT116, SW480, and HT29 colon cancer cells. *P < 0.05 versus NFs. All experiments were repeated three times, and representative results are shown here. (c, d) FGF-1 content in conditioned media was determined (c), and FGF-3 protein (d) was measured in cell lysates from NFs, PFs, CAFs, and HCT116, SW480, and HT29 colon cancer cells. Each value represents the mean ± SEM (n = 6). *P < 0.05 versus NFs. All experiments were repeated three times, and representative results are shown here. (e) Colon cancer cells were incubated with NF- or CAF-conditioned media for 48 h. The resulting cell lysates were subjected to co-immunoprecipitation and immunoblotting analyses with the indicated antibodies. All experiments were repeated three times, and representative results are shown here. (f) Colon cancer cells were incubated with NF-, PF-, or CAF-conditioned media for 48 h. The resulting lysates were subjected to immunoblotting with the indicated antibodies. The soluble MMP-7 was isolated from human colon cancer cell culture media. All experiments were repeated three times, and representative results are shown here.
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fig05: Cancer-associated fibroblasts (CAFs) predominantly secrete FGF1 and FGF3 and can upregulate the Mek/Erk pathway and MMP-7 through the activation of FGFR4. (a) mRNA levels of FGF-1, -2, -3, -7, EGF and HGF in NFs, PFs and CAFs were quantified through qRT-PCR. *P < 0.05 versus NFs. All experiments were repeated three times, and representative results are shown here. (b) The relative FGF1 and FGF3 mRNA levels in NFs, PFs, CAFs, and HCT116, SW480, and HT29 colon cancer cells. *P < 0.05 versus NFs. All experiments were repeated three times, and representative results are shown here. (c, d) FGF-1 content in conditioned media was determined (c), and FGF-3 protein (d) was measured in cell lysates from NFs, PFs, CAFs, and HCT116, SW480, and HT29 colon cancer cells. Each value represents the mean ± SEM (n = 6). *P < 0.05 versus NFs. All experiments were repeated three times, and representative results are shown here. (e) Colon cancer cells were incubated with NF- or CAF-conditioned media for 48 h. The resulting cell lysates were subjected to co-immunoprecipitation and immunoblotting analyses with the indicated antibodies. All experiments were repeated three times, and representative results are shown here. (f) Colon cancer cells were incubated with NF-, PF-, or CAF-conditioned media for 48 h. The resulting lysates were subjected to immunoblotting with the indicated antibodies. The soluble MMP-7 was isolated from human colon cancer cell culture media. All experiments were repeated three times, and representative results are shown here.

Mentions: Consistent with the in vivo observations, we found that the mRNA expression of FGF-1 and FGF-3 was remarkably increased in CAFs (Fig.5a). In addition to FGFs, CAFs also secreted other growth factors and chemokines, including SDF-1/CXCL12, VEGF, HGF, and CXCL14, into the tumor microenvironment for the promotion of tumourigenesis.6,10,21,22 However, the growth factors HGF, EGF, FGF-2, and FGF-7 showed no change in expression compared with isolated NFs and PFs (Fig.5a). Moreover, the expression of FGF-1 and FGF-3 in CAFs was abundant not only at the mRNA level but also at the protein level, in contrast to what was observed in colon cancer cells (Fig.5b–d), suggesting that FGF-1 and FGF-3 primarily act as autocrine mediators in CAFs, thereby contributing to the progression of colorectal cancer. Next, we cultured SW480, HCT116, and HT29 human colon cancer cells with CAF-, PF- and NF-conditioned media to examine the expression of FGFR4. Co-immunoprecipitation and immunoblotting analysis showed that FGFR4 phosphorylation (tyrosine kinase phosphorylation) was increased in cancer cells maintained in CAF-conditioned media compared with cells incubated in NF- or PF- conditioned media. However, total FGFR4 expression did not change (Fig.5e,f). The phosphorylation level of Mek/Erk was enhanced in cancer cells maintained in CAF-conditioned media compared with cells incubated in NF- or PF-conditioned media, although no apparent differences were observed in total Mek/Erk expression (Fig.5f). In addition, the protein expression and activation of MMP-7 were upregulated in the lysates and supernatants of SW480, HCT116, and HT29 cells after culture with CAF-conditioned media compared to culture with NF- or PF-conditioned media (Fig.5f).


FGF-1/-3/FGFR4 signaling in cancer-associated fibroblasts promotes tumor progression in colon cancer through Erk and MMP-7.

Bai YP, Shang K, Chen H, Ding F, Wang Z, Liang C, Xu Y, Sun MH, Li YY - Cancer Sci. (2015)

Cancer-associated fibroblasts (CAFs) predominantly secrete FGF1 and FGF3 and can upregulate the Mek/Erk pathway and MMP-7 through the activation of FGFR4. (a) mRNA levels of FGF-1, -2, -3, -7, EGF and HGF in NFs, PFs and CAFs were quantified through qRT-PCR. *P < 0.05 versus NFs. All experiments were repeated three times, and representative results are shown here. (b) The relative FGF1 and FGF3 mRNA levels in NFs, PFs, CAFs, and HCT116, SW480, and HT29 colon cancer cells. *P < 0.05 versus NFs. All experiments were repeated three times, and representative results are shown here. (c, d) FGF-1 content in conditioned media was determined (c), and FGF-3 protein (d) was measured in cell lysates from NFs, PFs, CAFs, and HCT116, SW480, and HT29 colon cancer cells. Each value represents the mean ± SEM (n = 6). *P < 0.05 versus NFs. All experiments were repeated three times, and representative results are shown here. (e) Colon cancer cells were incubated with NF- or CAF-conditioned media for 48 h. The resulting cell lysates were subjected to co-immunoprecipitation and immunoblotting analyses with the indicated antibodies. All experiments were repeated three times, and representative results are shown here. (f) Colon cancer cells were incubated with NF-, PF-, or CAF-conditioned media for 48 h. The resulting lysates were subjected to immunoblotting with the indicated antibodies. The soluble MMP-7 was isolated from human colon cancer cell culture media. All experiments were repeated three times, and representative results are shown here.
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fig05: Cancer-associated fibroblasts (CAFs) predominantly secrete FGF1 and FGF3 and can upregulate the Mek/Erk pathway and MMP-7 through the activation of FGFR4. (a) mRNA levels of FGF-1, -2, -3, -7, EGF and HGF in NFs, PFs and CAFs were quantified through qRT-PCR. *P < 0.05 versus NFs. All experiments were repeated three times, and representative results are shown here. (b) The relative FGF1 and FGF3 mRNA levels in NFs, PFs, CAFs, and HCT116, SW480, and HT29 colon cancer cells. *P < 0.05 versus NFs. All experiments were repeated three times, and representative results are shown here. (c, d) FGF-1 content in conditioned media was determined (c), and FGF-3 protein (d) was measured in cell lysates from NFs, PFs, CAFs, and HCT116, SW480, and HT29 colon cancer cells. Each value represents the mean ± SEM (n = 6). *P < 0.05 versus NFs. All experiments were repeated three times, and representative results are shown here. (e) Colon cancer cells were incubated with NF- or CAF-conditioned media for 48 h. The resulting cell lysates were subjected to co-immunoprecipitation and immunoblotting analyses with the indicated antibodies. All experiments were repeated three times, and representative results are shown here. (f) Colon cancer cells were incubated with NF-, PF-, or CAF-conditioned media for 48 h. The resulting lysates were subjected to immunoblotting with the indicated antibodies. The soluble MMP-7 was isolated from human colon cancer cell culture media. All experiments were repeated three times, and representative results are shown here.
Mentions: Consistent with the in vivo observations, we found that the mRNA expression of FGF-1 and FGF-3 was remarkably increased in CAFs (Fig.5a). In addition to FGFs, CAFs also secreted other growth factors and chemokines, including SDF-1/CXCL12, VEGF, HGF, and CXCL14, into the tumor microenvironment for the promotion of tumourigenesis.6,10,21,22 However, the growth factors HGF, EGF, FGF-2, and FGF-7 showed no change in expression compared with isolated NFs and PFs (Fig.5a). Moreover, the expression of FGF-1 and FGF-3 in CAFs was abundant not only at the mRNA level but also at the protein level, in contrast to what was observed in colon cancer cells (Fig.5b–d), suggesting that FGF-1 and FGF-3 primarily act as autocrine mediators in CAFs, thereby contributing to the progression of colorectal cancer. Next, we cultured SW480, HCT116, and HT29 human colon cancer cells with CAF-, PF- and NF-conditioned media to examine the expression of FGFR4. Co-immunoprecipitation and immunoblotting analysis showed that FGFR4 phosphorylation (tyrosine kinase phosphorylation) was increased in cancer cells maintained in CAF-conditioned media compared with cells incubated in NF- or PF- conditioned media. However, total FGFR4 expression did not change (Fig.5e,f). The phosphorylation level of Mek/Erk was enhanced in cancer cells maintained in CAF-conditioned media compared with cells incubated in NF- or PF-conditioned media, although no apparent differences were observed in total Mek/Erk expression (Fig.5f). In addition, the protein expression and activation of MMP-7 were upregulated in the lysates and supernatants of SW480, HCT116, and HT29 cells after culture with CAF-conditioned media compared to culture with NF- or PF-conditioned media (Fig.5f).

Bottom Line: The phosphorylation of FGFR4 enhanced the production of metalloproteinase (MMP)-7 and mitogen-activated protein kinase kinase (Mek)/extracellular signal-regulated kinase (Erk), which was accompanied by excessive vessel generation and cell proliferation.The administration of FGF-1/-3-neutralizing antibodies or the treatment of cells with FGFR4 siRNA or the FGFR4 inhibitor PD173074 markedly suppressed colon cancer cell proliferation and neovascularization.The inhibition of the FGF signaling pathway may be a useful strategy for the treatment of colon cancer.

View Article: PubMed Central - PubMed

Affiliation: Cancer Research Institute, Fudan University Shanghai Cancer Center, Shanghai, China.

No MeSH data available.


Related in: MedlinePlus