Cationic lipid mediated transfer of c-abl and bcr antisense oligonucleotides to immature normal myeloid cells: uptake, biological effects and modulation of gene expression.
Uptake and biochemical and biological effects of antisense oligodeoxynucleotides (ODN) specific for c-abl and bcr genes were studied in normal immature myeloid cells. CD34-positive cells were purified by positive and negative selection and cultured in liquid culture for 7 days. These cells were then incubated with ODNs, either alone or in combination with cationic lipids. The uptake of ODNs was enhanced by the use of cationic lipids. In addition, very low concentrations of ODNs in combination with cationic lipids were capable of specifically inhibiting the expression of the c-abl gene. In contrast, no effects were seen on the expression of bcr. However, despite the effective blocking of c-abl expression, no changes in cellular growth patterns were observed in liquid culture or in a colony-forming assay. We conclude tht the use of cationic lipids might enhance the gene-regulatory effects of ODNs by increasing their uptake into normal hematopoietic cells.
Albrecht
T
T
Division of Hematology, Third Department of Internal Medicine, Medical School of the Johannes, University of Mainz, Germany.
Schwab
R
R
Peschel
C
C
Engels
H J
HJ
Fischer
T
T
Huber
C
C
Aulitzky
W E
WE
D018952:Antigens, CD34; D001483:Base Sequence; D001692:Biological Transport / Q000187:drug effects; D002411:Cation Exchange Resins / Q000494:pharmacology*; D002412:Cations; D002463:Cell Membrane Permeability / Q000187:drug effects; D002474:Cell-Free System; D002478:Cells, Cultured; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003864:Depression, Chemical; D005786:Gene Expression Regulation / Q000187:drug effects*; D006412:Hematopoietic Stem Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D008055:Lipids / Q000494:pharmacology*; D008081:Liposomes; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage* / Q000494:pharmacology; D015513:Oncogene Proteins / Q000096:biosynthesis / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011505:Protein-Tyrosine Kinases; D011518:Proto-Oncogene Proteins; D016315:Proto-Oncogene Proteins c-abl / Q000096:biosynthesis / Q000235:genetics*; D051562:Proto-Oncogene Proteins c-bcr; D012334:RNA, Neoplasm / Q000037:antagonists & inhibitors; D016914:Ribonuclease H / Q000378:metabolism; D014162:Transfection / Q000379:methods*
D018952:Antigens, CD34; D002411:Cation Exchange Resins; D002412:Cations; D008055:Lipids; C086724:Lipofectamine; D008081:Liposomes; D016376:Oligonucleotides, Antisense; D015513:Oncogene Proteins; D011518:Proto-Oncogene Proteins; D012334:RNA, Neoplasm; D011505:Protein-Tyrosine Kinases; D016315:Proto-Oncogene Proteins c-abl; C496613:BCR protein, human; D051562:Proto-Oncogene Proteins c-bcr; D016914:Ribonuclease H