Breast Cancer Research and Treatment

, Volume 128, Issue 3, pp 691–701 | Cite as

Anti-tissue factor short hairpin RNA inhibits breast cancer growth in vivo

  • J. E. Bluff
  • M. Amarzguioui
  • J. Slattery
  • M. W. R. Reed
  • N. J. Brown
  • C. A. Staton
Preclinical study

Abstract

In breast cancer, there is a correlation between tissue factor (TF) expression, angiogenesis and disease progression. TF stimulates tumour angiogenesis, in part, through up-regulation of vascular endothelial growth factor (VEGF). Therefore, this study aimed to establish whether TF stimulates angiogenesis and tumour progression directly and independent of VEGF up-regulation. Initially, the effects of TF and VEGF were assessed on endothelial cell migration (Boyden chamber) and differentiation (tubule formation on Matrigel). Subsequently, MDA-MB-436 breast cancer cells, which produce high levels of both TF and VEGF (western blot analysis), were established in vivo, following which tumours were treated three times per week for 3 weeks with intra-tumoural injections of either anti-VEGF siRNA, anti-TF shRNA, the two treatments combined, or relevant controls. Both VEGF and TF significantly stimulated endothelial cell migration and tubule formation (P < 0.02). Breast cancer xenografts (MDA-MB-436) treated with TF or VEGF-specific agents demonstrated significant inhibition in tumour growth (VEGFsiRNA 61%; final volume: 236.2 ± 23.2 mm3 vs TFshRNA 89%; 161.9 ± 17.4 mm3 vs combination 93%; 136.3 ± 9.2 mm3 vs control 400.4 ± 32.7 mm3; P < 0.005). Microvessel density (MVD), a measure of angiogenesis, was also significantly inhibited in all groups (MVD in control = 29 ± 2.9; TFshRNA = 18 ± 1.1; VEGFsiRNA = 16.7 ± 1.5; both = 12 ± 2.1; P < 0.004), whereas the proliferative index of the tumours was only reduced in the TFshRNA-treated groups (control = 0.51 ± 0.011; TFshRNA = 0.41 ± 0.014; VEGFsiRNA = 0.49 ± 0.013; both = 0.41 ± 0.004; P < 0.008). These data suggest that TF has a direct effect on primary breast cancer growth and angiogenesis, and that specific inhibition of the TF-signalling pathway has potential for the treatment of primary breast cancer.

Keywords

Angiogenesis Breast cancer Haemostasis Tissue factor VEGF 

Abbreviations

CCS

Cumulative Chalkley score

GF

Growth factor

HuDMEC

Human dermal microvascular endothelial cells

MVD

Microvessel density

PECAM

Platelet endothelial cell adhesion molecule

TF

Tissue factor

VEGF

Vascular endothelial growth factor

Supplementary material

10549_2010_1149_MOESM1_ESM.tif (17 mb)
Supplementary material 1 (TIFF 17377 kb)

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Copyright information

© Springer Science+Business Media, LLC. 2010

Authors and Affiliations

  • J. E. Bluff
    • 1
  • M. Amarzguioui
    • 2
  • J. Slattery
    • 1
  • M. W. R. Reed
    • 1
  • N. J. Brown
    • 1
  • C. A. Staton
    • 1
  1. 1.Microcirculation Research Group, Academic Unit of Surgical Oncology, Department of Oncology, Faculty of Medicine, Dentistry and HealthUniversity of Sheffield Medical SchoolSheffieldUK
  2. 2.siRNAsenseUniversity of OsloOsloNorway

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