Molecular and Cellular Biochemistry

, Volume 308, Issue 1–2, pp 161–168

RNA interference (RNAi)-mediated vascular endothelial growth factor-C (VEGF-C) reduction interferes with lymphangiogenesis and enhances Epirubicin sensitivity of breast cancer cells

  • Ping Sun
  • Jie Gao
  • Yan-Li Liu
  • Lu-Wan Wei
  • Li-Ping Wu
  • Zhi-Yu Liu
Article

Abstract

It has been reported that over-expression of vascular endothelial growth factor-C (VEGF-C) in tumors leads to increased lymphangiogenesis and resistance to chemotherapy. Therefore, we hypothesized that VEGF-C would be a good molecular target for cancer gene therapy. In this study, we silenced the expression of VEGF-C with the highly specific post-transcriptional suppression of RNA interference (RNAi) in human breast cancer MCF-7 cell line. The expression of VEGF-C was examined by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), and the effect of plasmid on human lymphatic endothelial cells (HLECs) in vitro was analyzed by migration and 3-(4, 5-dimethylt-hiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The sensitivity to anticancer agents was evaluated by MTT and apoptosis assay, and apoptosis-related genes bcl-2/bax ratio was determined by Western Blotting. Results showed that of three siRNA-expressing vectors, P-1/siRNA most significantly suppressed the expression of VEGF-C mRNA and protein (38.1% of control and 117.8 ± 24.2 pg/ml, respectively) and interfered with proliferation and migration of HLECs in vitro. Moreover, transfection of VEGF-C/siRNA combined with Epirubicin markedly decreased breast cancer cells viability, reaching up to 38.5%, and increased apoptosis rate from 13.1% to 38.9%, as determined by decrease of bcl-2/bax ratio. In summary, VEGF-C would be a good molecular target, and a combination of Epirubicin and RNAi targeting VEGF-C could be an effective means for suppressing lymphatic metastasis and enhancing chemosensitivity of human breast cancer cells.

Keywords

Breastcancer Vascular endothelial growth factor-C (VEGF-C) RNA interference (RNAi) Human lymphatic endothelial cells (HLECs) Epirubicin 

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Copyright information

© Springer Science+Business Media, LLC. 2007

Authors and Affiliations

  • Ping Sun
    • 1
    • 2
  • Jie Gao
    • 1
  • Yan-Li Liu
    • 1
  • Lu-Wan Wei
    • 1
  • Li-Ping Wu
    • 1
  • Zhi-Yu Liu
    • 1
  1. 1.Department of Anatomy, School of MedicineShandong UniversityJiNanP.R. China
  2. 2.Department of AnatomyMuDanJiang Medical CollegeMuDanJiangP.R. China

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