Transfer of antisense oligodeoxynucleotides against endothelin receptors A and B into human coronary smooth muscle cells and endothelial cells by apolipoprotein E peptide: an in vitro study

Circ J. 2002 Nov;66(11):1054-6. doi: 10.1253/circj.66.1054.

Abstract

Antisense oligodeoxynucleotides (AS-ODNs) are a new generation of therapeutic agents for gene therapy. To develop a new approach in regulating the expression of endothelin (ET) receptor, N,N-dipalmitylglycyl-apolipoprotein E (129-169) peptide (dpGapoE), an efficient gene delivery system, was used to transfect phosphorothioated AS-ODNs against nucleotides of human ET type A (ETA) receptors in human coronary smooth muscle cells (HCSMCs) and type B (ETB) receptors in human coronary endothelial cells (HCECs). After transfection, translocation to the nuclei and concentration in nuclear structures were observed in approximately 40% of HCSMCs and 60% of HCECs, respectively, at 48 h by fluorescence microscopy. Both the cellular ETA mRNA concentration in HCSMCs and ETB mRNA concentration in HCECs significantly declined. This approach may enable gene regulation in vivo and could be used to regulate vascular tone and constriction through ET receptors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apolipoproteins E / chemistry
  • Apolipoproteins E / pharmacokinetics
  • Cell Culture Techniques
  • Coronary Vessels / cytology*
  • Coronary Vessels / metabolism
  • Endothelin Receptor Antagonists*
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / metabolism
  • Genetic Therapy / methods
  • Humans
  • Muscle, Smooth, Vascular / cytology
  • Muscle, Smooth, Vascular / metabolism
  • Myocytes, Smooth Muscle / metabolism
  • Oligonucleotides, Antisense / pharmacokinetics*
  • Oligonucleotides, Antisense / pharmacology
  • Peptide Fragments / pharmacokinetics
  • RNA, Messenger / analysis
  • RNA, Messenger / drug effects
  • Receptor, Endothelin A
  • Receptor, Endothelin B
  • Receptors, Endothelin / genetics
  • Transfection

Substances

  • Apolipoproteins E
  • Endothelin Receptor Antagonists
  • Oligonucleotides, Antisense
  • Peptide Fragments
  • RNA, Messenger
  • Receptor, Endothelin A
  • Receptor, Endothelin B
  • Receptors, Endothelin