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Lenvatinib in combination with golvatinib overcomes hepatocyte growth factor pathway-induced resistance to vascular endothelial growth factor receptor inhibitor.

Nakagawa T, Matsushima T, Kawano S, Nakazawa Y, Kato Y, Adachi Y, Abe T, Semba T, Yokoi A, Matsui J, Tsuruoka A, Funahashi Y - Cancer Sci. (2014)

Bottom Line: Here, we explored the effect of the HGF/Met signaling pathway and its inhibitors on resistance to lenvatinib, a VEGFR inhibitor.Lenvatinib potently inhibited the growth of HUVECs induced by VEGF alone, but cells induced by VEGF plus HGF showed lenvatinib resistance.This HGF-induced resistance was cancelled when the Met inhibitor, golvatinib, was added with lenvatinib.

View Article: PubMed Central - PubMed

Affiliation: Tsukuba Research Laboratory, Eisai Co., Ltd., Tsukuba, Japan.

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Related in: MedlinePlus

Effect of combined hepatocyte growth factor (HGF)/Met signaling pathway inhibitors on HGF-induced resistance to lenvatinib inhibition of HUVEC proliferation in vitro. (a) HUVECs were incubated with various concentrations of lenvatinib in the presence of vascular endothelial growth factor (VEGF) or VEGF plus HGF. (b) HUVECs were incubated with VEGF plus HGF for 3 days, then treated with lenvatinib (5 nM), golvatinib (30 nM), or both. Data represent means ± SD. *P < 0.05 versus control; **P < 0.05 versus the indicated group. (c) HUVECs were treated with DMSO control, lenvatinib (10 nM), golvatinib (50 nM), or both lenvatinib and golvatinib for 1 h. Additionally, cells were stimulated with medium, VEGF (20 ng/mL), HGF (30 ng/mL), or both VEGF and HGF for 10 min. The cell lysates were then harvested and the indicated proteins were analyzed by Western blotting. (d) HUVECs were incubated with or without (control) conditioned medium from SEKI human melanoma cells for 3 days. (e) HUVECs were incubated with conditioned medium from SEKI cells for 3 days, then treated with lenvatinib (10 nM) alone or in combination with either human HGF neutralizing Ab (10 μg/mL) or golvatinib (50 nM). Data represent means ± SD. *P < 0.05 versus the indicated group.
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fig02: Effect of combined hepatocyte growth factor (HGF)/Met signaling pathway inhibitors on HGF-induced resistance to lenvatinib inhibition of HUVEC proliferation in vitro. (a) HUVECs were incubated with various concentrations of lenvatinib in the presence of vascular endothelial growth factor (VEGF) or VEGF plus HGF. (b) HUVECs were incubated with VEGF plus HGF for 3 days, then treated with lenvatinib (5 nM), golvatinib (30 nM), or both. Data represent means ± SD. *P < 0.05 versus control; **P < 0.05 versus the indicated group. (c) HUVECs were treated with DMSO control, lenvatinib (10 nM), golvatinib (50 nM), or both lenvatinib and golvatinib for 1 h. Additionally, cells were stimulated with medium, VEGF (20 ng/mL), HGF (30 ng/mL), or both VEGF and HGF for 10 min. The cell lysates were then harvested and the indicated proteins were analyzed by Western blotting. (d) HUVECs were incubated with or without (control) conditioned medium from SEKI human melanoma cells for 3 days. (e) HUVECs were incubated with conditioned medium from SEKI cells for 3 days, then treated with lenvatinib (10 nM) alone or in combination with either human HGF neutralizing Ab (10 μg/mL) or golvatinib (50 nM). Data represent means ± SD. *P < 0.05 versus the indicated group.

Mentions: We first examined the effects of VEGF and HGF in HUVECs, an in vitro angiogenesis model. Addition of either VEGF or HGF significantly promoted cell proliferation and network formation of capillary-like vessels of HUVECs compared to that observed with vehicle alone (control) (Fig.1b,c). Costimulation with VEGF and HGF enhanced proliferation and tube formation of HUVECs to a significantly greater extent than that obtained with either single treatment (Fig.1b,c), and the networks formed after costimulation were more dense and intricately branched than those induced by either single factor (Fig.1d). We next examined the proliferation-inhibiting activity of lenvatinib, which is a potent VEGFR inhibitor, but not a Met inhibitor (Fig.1a). In HUVECs stimulated with VEGF alone, lenvatinib inhibited proliferation at IC50 1.6 nM and reached a plateau of approximately 80% inhibition (Fig.2a). In comparison, treatment with VEGF plus HGF showed decreased inhibitory activity of lenvatinib (IC50, 5.5 nM; plateau of approximately 60% inhibition), indicating that the HUVECs to which HGF had been added were resistant to lenvatinib.


Lenvatinib in combination with golvatinib overcomes hepatocyte growth factor pathway-induced resistance to vascular endothelial growth factor receptor inhibitor.

Nakagawa T, Matsushima T, Kawano S, Nakazawa Y, Kato Y, Adachi Y, Abe T, Semba T, Yokoi A, Matsui J, Tsuruoka A, Funahashi Y - Cancer Sci. (2014)

Effect of combined hepatocyte growth factor (HGF)/Met signaling pathway inhibitors on HGF-induced resistance to lenvatinib inhibition of HUVEC proliferation in vitro. (a) HUVECs were incubated with various concentrations of lenvatinib in the presence of vascular endothelial growth factor (VEGF) or VEGF plus HGF. (b) HUVECs were incubated with VEGF plus HGF for 3 days, then treated with lenvatinib (5 nM), golvatinib (30 nM), or both. Data represent means ± SD. *P < 0.05 versus control; **P < 0.05 versus the indicated group. (c) HUVECs were treated with DMSO control, lenvatinib (10 nM), golvatinib (50 nM), or both lenvatinib and golvatinib for 1 h. Additionally, cells were stimulated with medium, VEGF (20 ng/mL), HGF (30 ng/mL), or both VEGF and HGF for 10 min. The cell lysates were then harvested and the indicated proteins were analyzed by Western blotting. (d) HUVECs were incubated with or without (control) conditioned medium from SEKI human melanoma cells for 3 days. (e) HUVECs were incubated with conditioned medium from SEKI cells for 3 days, then treated with lenvatinib (10 nM) alone or in combination with either human HGF neutralizing Ab (10 μg/mL) or golvatinib (50 nM). Data represent means ± SD. *P < 0.05 versus the indicated group.
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fig02: Effect of combined hepatocyte growth factor (HGF)/Met signaling pathway inhibitors on HGF-induced resistance to lenvatinib inhibition of HUVEC proliferation in vitro. (a) HUVECs were incubated with various concentrations of lenvatinib in the presence of vascular endothelial growth factor (VEGF) or VEGF plus HGF. (b) HUVECs were incubated with VEGF plus HGF for 3 days, then treated with lenvatinib (5 nM), golvatinib (30 nM), or both. Data represent means ± SD. *P < 0.05 versus control; **P < 0.05 versus the indicated group. (c) HUVECs were treated with DMSO control, lenvatinib (10 nM), golvatinib (50 nM), or both lenvatinib and golvatinib for 1 h. Additionally, cells were stimulated with medium, VEGF (20 ng/mL), HGF (30 ng/mL), or both VEGF and HGF for 10 min. The cell lysates were then harvested and the indicated proteins were analyzed by Western blotting. (d) HUVECs were incubated with or without (control) conditioned medium from SEKI human melanoma cells for 3 days. (e) HUVECs were incubated with conditioned medium from SEKI cells for 3 days, then treated with lenvatinib (10 nM) alone or in combination with either human HGF neutralizing Ab (10 μg/mL) or golvatinib (50 nM). Data represent means ± SD. *P < 0.05 versus the indicated group.
Mentions: We first examined the effects of VEGF and HGF in HUVECs, an in vitro angiogenesis model. Addition of either VEGF or HGF significantly promoted cell proliferation and network formation of capillary-like vessels of HUVECs compared to that observed with vehicle alone (control) (Fig.1b,c). Costimulation with VEGF and HGF enhanced proliferation and tube formation of HUVECs to a significantly greater extent than that obtained with either single treatment (Fig.1b,c), and the networks formed after costimulation were more dense and intricately branched than those induced by either single factor (Fig.1d). We next examined the proliferation-inhibiting activity of lenvatinib, which is a potent VEGFR inhibitor, but not a Met inhibitor (Fig.1a). In HUVECs stimulated with VEGF alone, lenvatinib inhibited proliferation at IC50 1.6 nM and reached a plateau of approximately 80% inhibition (Fig.2a). In comparison, treatment with VEGF plus HGF showed decreased inhibitory activity of lenvatinib (IC50, 5.5 nM; plateau of approximately 60% inhibition), indicating that the HUVECs to which HGF had been added were resistant to lenvatinib.

Bottom Line: Here, we explored the effect of the HGF/Met signaling pathway and its inhibitors on resistance to lenvatinib, a VEGFR inhibitor.Lenvatinib potently inhibited the growth of HUVECs induced by VEGF alone, but cells induced by VEGF plus HGF showed lenvatinib resistance.This HGF-induced resistance was cancelled when the Met inhibitor, golvatinib, was added with lenvatinib.

View Article: PubMed Central - PubMed

Affiliation: Tsukuba Research Laboratory, Eisai Co., Ltd., Tsukuba, Japan.

Show MeSH
Related in: MedlinePlus