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Foretinib inhibits angiogenesis, lymphangiogenesis and tumor growth of pancreatic cancer in vivo by decreasing VEGFR-2/3 and TIE-2 signaling.

Chen HM, Tsai CH, Hung WC - Oncotarget (2015)

Bottom Line: We found Foretinib decreased basal- and HGF-induced c-MET activity at low concentrations.Foretinib inhibited VEGF-A, VEGF-C and Angiopoetin-2 (ANG-2)-stimulated tube formation and sprouting of LECs by reducing VEGFR-2, VEGFR-3 and TIE-2 activation and increased apoptosis of LECs.Additionally, Foretinib inhibited angiogenesis and lymphangiogenesis more significantly and exhibited low detrimental effect in orthotopic animal study.

View Article: PubMed Central - PubMed

Affiliation: National Institute of Cancer Research, National Health Research Institutes, Tainan 704, Taiwan, Republic of China.

ABSTRACT
Foretinib, a multiple kinase inhibitor undergoing clinical trials, could suppress the activity of hepatocyte growth factor (HGF) receptor c-MET and vascular endothelial growth factor receptor-2 (VEGFR-2). In addition, Foretinib may inhibit two critical lymphangiogenic signaling receptors VEGFR-3 and TIE-2. However, the effect of Foretinib on lymphatic endothelial cells (LECs) in vitro and lymphangiogenesis in vivo is still unknown. We found Foretinib decreased basal- and HGF-induced c-MET activity at low concentrations. However, Foretinib only reduced the proliferation of pancreatic cancer cells at high concentration reflecting the intrinsic chemoresistance of pancreatic cancer cells. Foretinib inhibited VEGF-A, VEGF-C and Angiopoetin-2 (ANG-2)-stimulated tube formation and sprouting of LECs by reducing VEGFR-2, VEGFR-3 and TIE-2 activation and increased apoptosis of LECs. In xenograft animal study, Foretinib suppressed tumor growth by inhibiting proliferation, angiogenesis and lymphangiogenesis. Additionally, Foretinib inhibited angiogenesis and lymphangiogenesis more significantly and exhibited low detrimental effect in orthotopic animal study. Collectively, we suggested that Foretinib simultaneously inhibits cancer cells and LECs to reduce pancreatic tumor growth in vivo and demonstrated for the first time that Foretinib suppresses angiogenesis and lymphangiogenesis by blocking VEGFR-2/3 and TIE-2 signaling.

No MeSH data available.


Related in: MedlinePlus

Foretinib inhibits VEGF-C and ANG-2-induced tube formation and spheroid sprouting of LECsA. Tube formation assay was done as described in Materials and Methods. LECs were pre-treated with indicated concentrations of Foretinib for 20 min, stimulated with VEGF-C and seeded on pre-coated Matrigel for 4 h. The number of nodes, tube area and tube length formed by LECs was measured. Values were Mean ± SEM of at least three separate experiments. B. The LEC spheroids were suspended in collagen gel and incubated in medium containing indicated concentrations of Foretinib and co-incubated with VEGF-C for 24 h. The number of sprouts, cumulative sprouting length and average sprouting length was measured. Values were Mean ± SEM of four separate experiments. C. Effect of Foretinib on ANG-2-induced tube formation was assayed as described in (A). D. Effect of Foretinib on ANG-2-induced tube formation was assayed as described in (B). Statistical analysis was determined by One-Way ANOVA, followed by Dunnett test. *P < 0.05.
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Figure 2: Foretinib inhibits VEGF-C and ANG-2-induced tube formation and spheroid sprouting of LECsA. Tube formation assay was done as described in Materials and Methods. LECs were pre-treated with indicated concentrations of Foretinib for 20 min, stimulated with VEGF-C and seeded on pre-coated Matrigel for 4 h. The number of nodes, tube area and tube length formed by LECs was measured. Values were Mean ± SEM of at least three separate experiments. B. The LEC spheroids were suspended in collagen gel and incubated in medium containing indicated concentrations of Foretinib and co-incubated with VEGF-C for 24 h. The number of sprouts, cumulative sprouting length and average sprouting length was measured. Values were Mean ± SEM of four separate experiments. C. Effect of Foretinib on ANG-2-induced tube formation was assayed as described in (A). D. Effect of Foretinib on ANG-2-induced tube formation was assayed as described in (B). Statistical analysis was determined by One-Way ANOVA, followed by Dunnett test. *P < 0.05.

Mentions: We next investigated the effect of Foretinib on lymphangiogenesis induced by two pro-lymphangiogenic factors VEGF-C and ANG-2 because Foretinib could inhibit their cognate receptors VEGFR-3 and TIE-2. We measured the number of junctions in the tube network (number of nodes), the linear length of the tubes that connect with at least one node (tube length) and the area of tube network enclosed by tube (tube area). As shown in Fig. 2A, VEGF-C significantly increased the node number, tube length and tube area of LECs and Foretinib at the concentrations of 0.1 and 1 μM inhibited the effect of VEGF-C. In the spheroid sprout assay, spheres of LECs were embedded in a three-dimensional collagen gel and the sprouts induced by VEGF-C were counted. VEGF-C increased the sprouting of LECs spheroid and the increase was inhibited by Foretinib (Fig. 3B). ANG-2 also increased the node number, tube length and tube area of LECs which was inhibited by 1 μM of Foretinib. However, Foretinib at 0.1 μM only significantly decreased the tube area but not the node number and tube length (Fig. 2C). In the spheroid sprout assay, ANG-2 increased the number of sprouts, accumulated length per spheroid and average sprouting length and the increase was also inhibited by Foretinib (Fig. 2D).


Foretinib inhibits angiogenesis, lymphangiogenesis and tumor growth of pancreatic cancer in vivo by decreasing VEGFR-2/3 and TIE-2 signaling.

Chen HM, Tsai CH, Hung WC - Oncotarget (2015)

Foretinib inhibits VEGF-C and ANG-2-induced tube formation and spheroid sprouting of LECsA. Tube formation assay was done as described in Materials and Methods. LECs were pre-treated with indicated concentrations of Foretinib for 20 min, stimulated with VEGF-C and seeded on pre-coated Matrigel for 4 h. The number of nodes, tube area and tube length formed by LECs was measured. Values were Mean ± SEM of at least three separate experiments. B. The LEC spheroids were suspended in collagen gel and incubated in medium containing indicated concentrations of Foretinib and co-incubated with VEGF-C for 24 h. The number of sprouts, cumulative sprouting length and average sprouting length was measured. Values were Mean ± SEM of four separate experiments. C. Effect of Foretinib on ANG-2-induced tube formation was assayed as described in (A). D. Effect of Foretinib on ANG-2-induced tube formation was assayed as described in (B). Statistical analysis was determined by One-Way ANOVA, followed by Dunnett test. *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4558127&req=5

Figure 2: Foretinib inhibits VEGF-C and ANG-2-induced tube formation and spheroid sprouting of LECsA. Tube formation assay was done as described in Materials and Methods. LECs were pre-treated with indicated concentrations of Foretinib for 20 min, stimulated with VEGF-C and seeded on pre-coated Matrigel for 4 h. The number of nodes, tube area and tube length formed by LECs was measured. Values were Mean ± SEM of at least three separate experiments. B. The LEC spheroids were suspended in collagen gel and incubated in medium containing indicated concentrations of Foretinib and co-incubated with VEGF-C for 24 h. The number of sprouts, cumulative sprouting length and average sprouting length was measured. Values were Mean ± SEM of four separate experiments. C. Effect of Foretinib on ANG-2-induced tube formation was assayed as described in (A). D. Effect of Foretinib on ANG-2-induced tube formation was assayed as described in (B). Statistical analysis was determined by One-Way ANOVA, followed by Dunnett test. *P < 0.05.
Mentions: We next investigated the effect of Foretinib on lymphangiogenesis induced by two pro-lymphangiogenic factors VEGF-C and ANG-2 because Foretinib could inhibit their cognate receptors VEGFR-3 and TIE-2. We measured the number of junctions in the tube network (number of nodes), the linear length of the tubes that connect with at least one node (tube length) and the area of tube network enclosed by tube (tube area). As shown in Fig. 2A, VEGF-C significantly increased the node number, tube length and tube area of LECs and Foretinib at the concentrations of 0.1 and 1 μM inhibited the effect of VEGF-C. In the spheroid sprout assay, spheres of LECs were embedded in a three-dimensional collagen gel and the sprouts induced by VEGF-C were counted. VEGF-C increased the sprouting of LECs spheroid and the increase was inhibited by Foretinib (Fig. 3B). ANG-2 also increased the node number, tube length and tube area of LECs which was inhibited by 1 μM of Foretinib. However, Foretinib at 0.1 μM only significantly decreased the tube area but not the node number and tube length (Fig. 2C). In the spheroid sprout assay, ANG-2 increased the number of sprouts, accumulated length per spheroid and average sprouting length and the increase was also inhibited by Foretinib (Fig. 2D).

Bottom Line: We found Foretinib decreased basal- and HGF-induced c-MET activity at low concentrations.Foretinib inhibited VEGF-A, VEGF-C and Angiopoetin-2 (ANG-2)-stimulated tube formation and sprouting of LECs by reducing VEGFR-2, VEGFR-3 and TIE-2 activation and increased apoptosis of LECs.Additionally, Foretinib inhibited angiogenesis and lymphangiogenesis more significantly and exhibited low detrimental effect in orthotopic animal study.

View Article: PubMed Central - PubMed

Affiliation: National Institute of Cancer Research, National Health Research Institutes, Tainan 704, Taiwan, Republic of China.

ABSTRACT
Foretinib, a multiple kinase inhibitor undergoing clinical trials, could suppress the activity of hepatocyte growth factor (HGF) receptor c-MET and vascular endothelial growth factor receptor-2 (VEGFR-2). In addition, Foretinib may inhibit two critical lymphangiogenic signaling receptors VEGFR-3 and TIE-2. However, the effect of Foretinib on lymphatic endothelial cells (LECs) in vitro and lymphangiogenesis in vivo is still unknown. We found Foretinib decreased basal- and HGF-induced c-MET activity at low concentrations. However, Foretinib only reduced the proliferation of pancreatic cancer cells at high concentration reflecting the intrinsic chemoresistance of pancreatic cancer cells. Foretinib inhibited VEGF-A, VEGF-C and Angiopoetin-2 (ANG-2)-stimulated tube formation and sprouting of LECs by reducing VEGFR-2, VEGFR-3 and TIE-2 activation and increased apoptosis of LECs. In xenograft animal study, Foretinib suppressed tumor growth by inhibiting proliferation, angiogenesis and lymphangiogenesis. Additionally, Foretinib inhibited angiogenesis and lymphangiogenesis more significantly and exhibited low detrimental effect in orthotopic animal study. Collectively, we suggested that Foretinib simultaneously inhibits cancer cells and LECs to reduce pancreatic tumor growth in vivo and demonstrated for the first time that Foretinib suppresses angiogenesis and lymphangiogenesis by blocking VEGFR-2/3 and TIE-2 signaling.

No MeSH data available.


Related in: MedlinePlus