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Tumor Biology

, Volume 37, Issue 9, pp 12791–12803 | Cite as

Evodiamine exerts anti-tumor effects against hepatocellular carcinoma through inhibiting β-catenin-mediated angiogenesis

  • Le Shi
  • Fan Yang
  • Fei Luo
  • Yi Liu
  • Feng Zhang
  • Meijuan Zou
  • Qizhan Liu
Original Article

Abstract

Hepatocellular carcinoma (HCC) is a highly vascular tumor with high microvessel density and high levels of circulating vascular endothelial growth factor (VEGF). Thus, the angiogenesis pathway is an attractive therapeutic target for HCC. The anti-tumor effects of evodiamine, a quinolone alkaloid isolated from Euodia rutaecarpa (Juss.) Benth. (Rutaceae), were investigated in a mouse xenograft model using BALB/c nude mice, various HCC cell lines (HepG2, SMMC-7721, H22), and human umbilical vein endothelial cells (HUVECs). The effects of evodiamine on tumor volumes and weights, levels of tumor markers, angiogenesis in vivo and in vitro, cell viability, and cell migration and invasion were measured, and the mechanism through which its effects are achieved was investigated. Transcriptional regulation of VEGFa via interaction with β-catenin was established by luciferase activity assays and electrophoretic mobility shift assays. In a subcutaneous H22 xenograft model, evodiamine inhibited tumor growth and reduced serum tumor markers and the levels of β-catenin and VEGFa. It also blocked VEGF-induced angiogenesis in a Matrigel plug assay. Evodiamine suppressed cellular proliferation, invasion, and migration and inhibited tube formation of HUVECs. Moreover, in a concentration-dependent manner, evodiamine reduced the number of capillary sprouts from Matrigel-embedded rat thoracic aortic rings. Also, evodiamine suppressed various biomarkers of angiogenesis and the expression of β-catenin. Evodiamine decreased β-catenin levels activated by LiCl, which led to reduced expression of VEGFa. In addition, β-catenin interacted with VEGFa and transcriptionally regulated VEGFa, an effect inhibited by evodiamine in HCCs. Moreover, in an SMMC-7721 xenograft model, evodiamine suppressed tumor growth, various biomarkers of angiogenesis, and the levels of β-catenin and VEGFa. Evodiamine has anti-tumor effects on HCCs through inhibiting β-catenin, which interacts with and reduces VEGFa expression, thus inhibiting angiogenesis. These results indicate that evodiamine, which inhibits cellular invasion and migration and blocks angiogenesis, is a potential therapeutic agent for HCCs.

Keywords

Evodiamine Angiogenesis Hepatocellular carcinoma β-Catenin Vascular endothelial growth factor 

Abbreviations

HCC

Hepatocellular carcinoma

VEGF

Vascular endothelial growth factor

HUVECs

Human umbilical vein endothelial cells

DMEM

Dulbecco’s modified Eagle medium

FBS

Fetal bovine serum

MTT

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide

IHC

Immunohistochemical

DAB

3,3-Diaminobenzidine tetrahydrochloride

SDS-PAGE

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

PVDF

Polyvinylidene fluoride

BSA

Bovine serum albumin

HRP

Horseradish peroxidase

AFP

Alpha fetal protein

TSGF

Tumor-specific growth factor

Notes

Acknowledgments

The authors wish to thank Donald L. Hill (University of Alabama at Birmingham, USA), an experienced, English-speaking scientific editor for editing.

Compliance with ethical standards

Conflicts of interest

None

Funding

This work was supported by the Natural Science Foundations of China (81273114, 81302467, 81402959), the Postgraduate Innovation Project of Jiangsu province (CXZZ14_0421, CXZZ14_0951, and KYLX15_0974), and the Priority Academic Program Development of Jiangsu Higher Education Institutions (2014).

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  • Le Shi
    • 1
    • 2
    • 3
  • Fan Yang
    • 3
  • Fei Luo
    • 1
    • 2
  • Yi Liu
    • 1
    • 2
  • Feng Zhang
    • 3
  • Meijuan Zou
    • 4
  • Qizhan Liu
    • 1
    • 2
  1. 1.Institute of Toxicology, School of Public HealthNanjing Medical UniversityNanjingChina
  2. 2.The Key Laboratory of Modern Toxicology, Ministry of Education, School of Public HealthNanjing Medical UniversityNanjingChina
  3. 3.School of PharmacyNanjing University of Chinese MedicineNanjingChina
  4. 4.Department of Pharmacology, School of Basic Medical SciencesNanjing Medical UniversityNanjingChina

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