Skip to main content
main-content

01.12.2018 | Research | Ausgabe 1/2018 Open Access

Journal of Experimental & Clinical Cancer Research 1/2018

The dual blockade of MET and VEGFR2 signaling demonstrates pronounced inhibition on tumor growth and metastasis of hepatocellular carcinoma

Zeitschrift:
Journal of Experimental & Clinical Cancer Research > Ausgabe 1/2018
Autoren:
Yu Zhang, Xiaomei Gao, Ying Zhu, Dhruba Kadel, Haoran Sun, Jing Chen, Qin Luo, Haoting Sun, Luyu Yang, Jing Yang, Yuanyuan Sheng, Yan Zheng, Kejin Zhu, Qiongzhu Dong, Lunxiu Qin
Wichtige Hinweise

Electronic supplementary material

The online version of this article (https://​doi.​org/​10.​1186/​s13046-018-0750-2) contains supplementary material, which is available to authorized users.

Abstract

Background

The application of VEGF signaling inhibitors have been associated with more invasive or metastatic behavior of cancers including hepatocellular carcinoma (HCC). We explored the contribution of MET pathway to the enhanced HCC invasion and metastasis by VEGF signaling inhibition, and investigated the antitumor effects of NZ001, a novel dual inhibitor of MET and VEGFR2, in HCC.

Methods

Immunocompetent orthotopic mice model of hepal-6 was established to investigate the effects of either VEGF antibody alone or in combination with the selective MET inhibitor on tumor aggressiveness. The antitumor effects of NZ001 were examined in cultured HCC cells as well as in vivo models. MET gene amplification was determined by SNP 6.0 assay. MET/P-MET expression was detected by IHC.

Results

Selective VEGF signaling inhibition by VEGF antibody significantly reduced in vivo tumor growth of the orthotopic mice models, simultaneously also enhanced tumor invasion and metastasis, but inhibiting MET signaling attenuated this side-effect. Further study revealed that hypoxia caused by VEGF signaling inhibition induced HIF-1α nuclear accumulation, subsequently leading to elevated total-MET expression, and synergized with HGF in inducing invasion. NZ001, a novel dual inhibitor of MET and VEGFR2, markedly inhibited both tumor growth and metastasis of HCC, which showed obvious advantages over sorafenib in not inducing more invasive and metastatic behaviors. This effect is more pronounced in HCC with MET amplification and overexpression.

Conclusions

The activation of MET is responsible for the metastasis-promoting effects induced by VEGF inhibition. MET and VEGFR2 dual blockade, NZ001, has advantages over sorafenib in not inducing more invasive and metastatic behaviors; MET amplification and overexpression can be used to identify the subgroup of patients most likely to get the optimal benefit from NZ001 treatment.
Zusatzmaterial
Additional file 3: Figure S1. The E-cadherin, vimentin, CD34, HIF-1α, P-MET and total-MET expression levels in hepa1–6 orthotopic tumors after treatment with vehicle and VEGF antibody. Figure S2. The expression levels of E-cadherin, N-cadherin and vimentin were detected by Western blot in HCC cells starved overnight and treated with HGF (10 ng/ml) for 24 h. Figure S3. After 2-week treatment with VEGF antibody, PF-04217903 alone, and their combination, the mice were killed and the liver tissues were obtained. Figure S4. The E-cadherin, vimentin, CD34, HIF-1α, total-MET and P-MET expression levels in hepa1–6 orthotopic tumors after treatment with VEGF antibody and PF-04217903 alone, or their combination. Figure S5. The effect of NZ001 on VEGFR2 and MET signaling in HUVECs and HCC cells. Figure S6. NZ001 suppressed the HGF-induced EMT in HCC cells. Figure S7. Effects of NZ001 on the spontaneous and chemokine-induced invasion of HCC cell under normoxia and hypoxia condition. Figure S8. After 2-week treatment with sorafenib and NZ001, the mice were killed and the liver tissues were obtained. Figure S9. The total-MET, HIF-1α, CD34 and P-MET expression levels in hepa1–6 orthotopic tumors after treatment with VEGF antibody and PF-04217903 alone, or their combination. Figure S10. Effects of NZ001 on the metastasis of MHCC97H in nude mice. Figure S11. Effects of NZ001 on colony-formation of HCC cells. Figure S12. The association of MET exon 14 mutation determined by sanger sequencing with NZ001 sensitivity of 12 HCC cell lines. Figure S13. The score of MET and P-MET definition for IHC staining. Figure S14. The MET/P-MET/HGF expression in HCC cell lines. Figure S15. After 2-week treatment with different concentrations of NZ001, the mice were sacrificed and the tumors were obtained. Figure S16. IHC assay showed that MET phosphorylation was greatly inhibited after NZ001 treatment in MHCC-97H xenograft tumors. (DOCX 5327 kb)
Literatur
Über diesen Artikel

Weitere Artikel der Ausgabe 1/2018

Journal of Experimental & Clinical Cancer Research 1/2018 Zur Ausgabe