Total alkaloids of Rubus alceifolius Poir shows anti-angiogenic activity in vivo and in vitro

Integr Cancer Ther. 2014 Nov;13(6):520-8. doi: 10.1177/1534735414547109. Epub 2014 Aug 22.

Abstract

Total alkaloids is an active ingredient of the natural plant Rubus alceifolius Poir, commonly used for the treatment of various cancers. Antitumor effects may be mediated through anti-angiogenic mechanisms. As such, the goal of the present study was to investigate and evaluate the effect of total alkaloids in Rubus alceifolius Poir (TARAP) on tumor angiogenesis and investigate the underlying molecular mechanisms of TARAP action in vivo and in vitro. A chick embryo chorioallantoic membrane (CAM) assay was used to assess angiogenesis in vivo. An MTT assay was performed to determine the viability of human umbilical vein endothelial cells (HUVECs) with and without treatment. Cell cycle progression of HUVECs was examined by FACS analysis with propidium iodide staining. HUVEC migration was determined using a scratch wound method. Tube formation of HUVECs was assessed with an ECMatrix gel system, and mRNA and protein expression of VEGF-A in both HUVECs and HepG2 human hepatocellular carcinoma cells were examined by RT-PCR and ELISA, respectively. Our results showed that TARAP inhibited angiogenesis in the CAM model in vivo and inhibited HUVEC proliferation via blocking cell cycle G1 to S progression in a dose- and time-dependent manners in vitro. Moreover, TARAP inhibited HUVEC migration and tube formation and downregulated mRNA and protein expression of VEGF-A in both HepG2 cells and HUVECs. Our findings suggest that the anti-angiogenic activity of TARAP may partly contribute to its antitumor properties and may be valuable for the treatment of diseases involving pathologic angiogenesis such as cancer.

Keywords: Rubus alceifolius Poir; TARAP; hepatocellular carcinoma; human umbilical vein endothelial cell; tumor angiogenesis.

MeSH terms

  • Alkaloids / isolation & purification
  • Alkaloids / pharmacology*
  • Angiogenesis Inhibitors / isolation & purification
  • Angiogenesis Inhibitors / pharmacology*
  • Animals
  • Carcinoma, Hepatocellular / blood supply
  • Carcinoma, Hepatocellular / drug therapy
  • Cell Survival / drug effects
  • Chick Embryo
  • Chorioallantoic Membrane / blood supply
  • Chorioallantoic Membrane / drug effects
  • Enzyme-Linked Immunosorbent Assay
  • Gene Expression Regulation / drug effects
  • Hep G2 Cells
  • Human Umbilical Vein Endothelial Cells / drug effects
  • Humans
  • Liver Neoplasms / blood supply
  • Liver Neoplasms / drug therapy
  • Neovascularization, Pathologic / drug therapy*
  • Neovascularization, Pathologic / pathology
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Rubus / chemistry*
  • Vascular Endothelial Growth Factor A / genetics

Substances

  • Alkaloids
  • Angiogenesis Inhibitors
  • RNA, Messenger
  • Vascular Endothelial Growth Factor A