title: Inhibition of P210BCR/ABL Expression in K562 Cells by Electroporation with an Antisense Oligonucleotide.
abstract: We designed experiments to study the effects on P210BCR/ABL expression of introducing antisense oligonucleotides into K562 cells. We used two antisense oligonucleotides: one (AS1) is complementary to the first coding codon of the BCR/ABL mRNA and the two 5' and three 3' codons, and the other (AS2) to BCR coding codons 5 to 11 inclusive. To facilitate entry of the oligonucleotides the K562 cells were subjected to electroporation on three occasions at 24 hr intervals (0, 24 and 48 hr). P210BCR/ABL expression was assayed by in vivo phosphorylation followed by immune precipitation with a BCR antibody. Introduction of AS1 inhibited P210BCR/ABL expression at 72 and 96 hrs, whereas AS2 and the control oligonucleotide had no effect. AS1 also killed K562 cells. We conclude that selected antisense oligonucleotides can modify leukaemia-specific protein expression in K562 cells. This approach could prove valuable for purging CML bone marrow cells in vitro.
authors.affiliation: a MRC/LRF Leukaemia Unit, Hammersmith Hospital, London.
authors.forename: A S
authors.initials: AS
authors.lastname: Taj
authors.affiliation: a MRC/LRF Leukaemia Unit, Hammersmith Hospital, London.
authors.forename: P
authors.initials: P
authors.lastname: Martiat
authors.affiliation: b Imperial Cancer Research Fund, St. Bartholomew's Hospital, London, UK.
authors.forename: S
authors.initials: S
authors.lastname: Dhut
authors.affiliation: b Imperial Cancer Research Fund, St. Bartholomew's Hospital, London, UK.
authors.forename: T L
authors.initials: TL
authors.lastname: Chaplin
authors.affiliation: a MRC/LRF Leukaemia Unit, Hammersmith Hospital, London.
authors.forename: C
authors.initials: C
authors.lastname: Dowding
authors.affiliation: a MRC/LRF Leukaemia Unit, Hammersmith Hospital, London.
authors.forename: K H
authors.initials: KH
authors.lastname: Th'ng
authors.affiliation: b Imperial Cancer Research Fund, St. Bartholomew's Hospital, London, UK.
authors.forename: I
authors.initials: I
authors.lastname: Goldstein
authors.affiliation: c Whitehead Institute, Cambridge, USA.
authors.forename: G Q
authors.initials: GQ
authors.lastname: Daley
authors.affiliation: b Imperial Cancer Research Fund, St. Bartholomew's Hospital, London, UK.
authors.forename: B D
authors.initials: BD
authors.lastname: Young
authors.affiliation: a MRC/LRF Leukaemia Unit, Hammersmith Hospital, London.
authors.forename: J M
authors.initials: JM
authors.lastname: Goldman
keywords: Antisense oligonucleotides; electroporation