Comparing transfection efficiency and safety for antisense oligodeoxyribonucleotide between phospholipids-based microbubbles and liposomes

J Drug Target. 2006 Dec;14(10):687-93. doi: 10.1080/10611860600965849.

Abstract

Objective: To compare transfection efficiency and safety for antisense oligodeoxynucleotides (AS-ODNs) between two type of phospholipids-based vectors.

Methods: An AS-ODNs sequence HA824 combined with luciferase reporter plasmid was used. Under low intensity ultrasound (US), a breast cancer cell line SK-BR-3 was exposed to different concentration of microbubbles and liposomes. Transfection efficiency was detected by fluorescence microscopy. Cell viability was verified by propidium iodide assay. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the inhibitory effect of HA824 on HER-2 expression at mRNA level. Atomic force microscopy (AFM) scanning techniques was employed to observe the change of membrane pore size.

Results: AS-ODNs transfection efficiency showed an increasing tend with microbubble concentration, but not with liposome concentration. Maximum transfection efficiency with minimum cell viability was achieved under 2% microbubble concentration. Too strong sonoporation activity would enlarge membrane pores significantly and cause low cell viability.

Conclusion: US-mediated AS-ODNs transfection enhanced by phospholipids-based microbubbles represents an effective and safe avenue.

Publication types

  • Comparative Study

MeSH terms

  • Cell Line, Tumor
  • Cell Membrane / metabolism
  • Cell Membrane / ultrastructure
  • Cell Survival / drug effects
  • Drug Carriers
  • Excipients
  • Female
  • Gene Transfer Techniques
  • Genes, Reporter / genetics
  • Genes, erbB-2 / genetics
  • Humans
  • Liposomes
  • Luciferases / genetics
  • Microspheres
  • Oligodeoxyribonucleotides, Antisense / administration & dosage
  • Oligodeoxyribonucleotides, Antisense / adverse effects*
  • Oligodeoxyribonucleotides, Antisense / pharmacology*
  • Phospholipids
  • Porosity
  • Propidium
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Spectrophotometry, Atomic
  • Transfection

Substances

  • Drug Carriers
  • Excipients
  • Liposomes
  • Oligodeoxyribonucleotides, Antisense
  • Phospholipids
  • RNA, Messenger
  • Propidium
  • Luciferases