Inhibition of VEGF expression in A431 and MDA-MB-231 tumour cells by cationic lipid-mediated siRNA delivery

J Drug Target. 2012 May;20(4):347-54. doi: 10.3109/1061186X.2012.656645.

Abstract

In order to promote siRNA transfer in tumour cells, we used an original cationic lipid, synthesized in our laboratory, dimethyl-hydroxyethyl-aminopropane-carbamoyl-cholesterol (DMHAPC-Chol). Liposomes were prepared from this lipid and dioleoylphosphatidylethanolamine (DOPE) in equimolar proportion. Its transfecting capacity was evaluated using ELISA, cell cytometry, and RT-PCR in estimating the silencing effect of VEGF siRNA. This liposome efficiently delivered VEGF siRNA in two human cancer cell lines abundantly secreting VEGF, A431 and MDA-MB-231. Results showed that 50 nM of VEGF siRNA carried by DMHAPC-Chol/DOPE liposomes already silenced more than 90% of VEGF in these cells. A comparative study with two commercial carriers indicated that the inhibition induced by VEGF siRNA transported by cationic DMHAPC-Chol/DOPE liposomes was comparable to that induced by INTERFERin and better than lipofectamine 2000. Moreover, a transfection by a GFP plasmid followed by a GFP siRNA showed that DMHAPC-Chol/DOPE liposomes compared to lipofectamine were less efficient for plasmid but better for siRNA transport. Following one of our previous works concerning cell delivery of plasmid ( Percot et al., 2004 ), the main interest of results presented here resides in the double potential of DMHAPC-Chol/DOPE liposomes to deliver little-sized siRNA as well as large nucleic acids in cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cations
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cholesterol / analogs & derivatives*
  • Cholesterol / chemical synthesis
  • Cholesterol / chemistry
  • DNA / administration & dosage
  • DNA / genetics
  • Drug Carriers / chemical synthesis
  • Drug Carriers / chemistry*
  • Enzyme-Linked Immunosorbent Assay
  • Flow Cytometry
  • Gene Silencing
  • Green Fluorescent Proteins / genetics
  • Humans
  • Liposomes
  • Plasmids
  • RNA, Small Interfering* / administration & dosage
  • RNA, Small Interfering* / genetics
  • RNA, Small Interfering* / pharmacology
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transfection
  • Vascular Endothelial Growth Factor A / genetics*

Substances

  • Cations
  • Drug Carriers
  • Liposomes
  • RNA, Small Interfering
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • DNA
  • Cholesterol