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Gramine inhibits angiogenesis and induces apoptosis via modulation of TGF-β signalling in 7,12 dimethylbenz[a]anthracene (DMBA) induced hamster buccal pouch carcinoma

Phytomedicine. 2017 Sep 15:33:69-76. doi: 10.1016/j.phymed.2017.05.008. Epub 2017 Jun 9.

Abstract

Background: Transforming growth factor-β (TGF-β) and its receptors are considered as a novel target in cancer chemotherapy. Gramine, an indole alkaloid, possesses various pharmacological properties including antiproliferative and anticancer. However, the anti-angiogenic property remains unexplored.

Purpose: The present study was designed to evaluate the anti-angiogenic and apoptosis induction properties of gramine through inhibiting TGF-β on DMBA induced oral squamous cell carcinoma (OSCC) in the hamster buccal pouch (HBP).

Methods: The effects of gramine on TGF-β signalling in DMBA induced carcinogenic events such as angiogenesis and apoptosis were analysed by studying the mRNA expression using RT-PCR, protein expression by western blot and histopathological analysis using haematoxylin and eosin (H & E) staining.

Results: Gramine significantly inhibited phosphorylation and nuclear translocation of Smad2 and Smad4 by blocking activity of the TGFβ-RII, RI and activation of inhibitory Smad7. Gramine inhibited angiogenic markers such as MMP-2, MMP-9, HIF-1α, VEGF, and VEGF-R2 as well as increased TIMP-2 expression. Furthermore, gramine induced apoptosis in DMBA induced tumour bearing animals by up regulating the pro apoptotic proteins Bax, cytochrome C, apaf-1, caspase-9 caspase-3 and PARP.

Conclusion: In this study, we clearly demonstrated that gramine treatment diminishes angiogenesis and induces apoptosis in hamster buccal pouch (HBP) carcinogenesis by modulating TGF-β signals.

Keywords: Angiogenesis; Apoptosis; Gramine; Oral cancer; TGF-β.

MeSH terms

  • 9,10-Dimethyl-1,2-benzanthracene
  • Alkaloids / pharmacology*
  • Angiogenesis Inhibitors / pharmacology*
  • Animals
  • Apoptosis / drug effects*
  • Carcinoma, Squamous Cell / chemically induced
  • Carcinoma, Squamous Cell / drug therapy*
  • Caspase 3 / metabolism
  • Caspase 9 / metabolism
  • Cheek / pathology
  • Cricetinae
  • Indole Alkaloids
  • Male
  • Mesocricetus
  • Mouth Neoplasms / chemically induced
  • Mouth Neoplasms / drug therapy*
  • Neovascularization, Pathologic / drug therapy
  • Signal Transduction
  • Tissue Inhibitor of Metalloproteinase-2 / metabolism
  • Transforming Growth Factor beta / metabolism*

Substances

  • Alkaloids
  • Angiogenesis Inhibitors
  • Indole Alkaloids
  • Transforming Growth Factor beta
  • Tissue Inhibitor of Metalloproteinase-2
  • 9,10-Dimethyl-1,2-benzanthracene
  • Caspase 3
  • Caspase 9
  • gramine