Reduction of erbB2 gene product in mamma carcinoma cell lines by erbB2 mRNA-specific and tyrosine kinase consensus phosphorothioate antisense oligonucleotides.
A considerable reduction of up to 75% at the protein level of the erbB2 gene product was observed using phosphorothioate antisense oligonucleotides directed against specific sequences of the erbB2 mRNA. Antisense oligonucleotides used were the 14-mer (translation start region) and the 17-mer (3' translated region) all-phosphorothioate oligonucleotides (S-ODNs). Sense or random sequences were used as a control. The greatest reduction at the erbB2 protein level was obtained after a 24h incubation with a single dose of 2 microM antisense S-ODN. The erbB2-mRNA-specific and the tyrosine kinase consensus antisense S-ODNs were comparably effective in inhibiting erbB2 expression. Addition of the transfection reagent DOTAP diminished the efficiency of erbB2 protein reduction by antisense S-ODNs after an incubation period of 24h but was more effective after 48h compared to the application of antisense S-ODNs alone.
Bertram
J
J
Dept. of Hematology/Onc. Univ. Clinics, Göttingen, FRG.
Killian
M
M
Brysch
W
W
Schlingensiepen
K H
KH
Kneba
M
M
D001483:Base Sequence; D001943:Breast Neoplasms; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D066246:ErbB Receptors / Q000096:biosynthesis* / Q000235:genetics; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D063086:Organothiophosphates; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis*; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis* / Q000235:genetics; D011519:Proto-Oncogenes; D012333:RNA, Messenger / Q000096:biosynthesis*; D018719:Erb-b2 Receptor Tyrosine Kinases; D014407:Tumor Cells, Cultured
D066246:ErbB Receptors; D016376:Oligonucleotides, Antisense; D063086:Organothiophosphates; D011505:Protein-Tyrosine Kinases; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D018719:Erb-b2 Receptor Tyrosine Kinases