Previously described cell membrane transport mechanisms are unable to
account completely for oligodeoxynucleotide cellular uptake. These cha
rged macromolecules enter cells by an incompletely defined mechanism a
nd downregulate gene expression in either the cytoplasm or nucleus. Th
us, the goal bf this research was to study the mechanism of phosphodie
ster oligonucleotide cellular uptake in Rauscher Red 5-1.5 erythroleuk
emia cells. An antisense c-myc oligodeoxynucleotide (21 bases) demonst
rated biological activity in these cells using two types of proliferat
ion assays and Northern blot analysis, and was internalized as visuali
zed by confocal laser microscopy. Oligonucleotide uptake appeared to b
e a complex process consisting of surface binding and internalization.
Cellular internalization accounted for up to 40% of total uptake and
was partially dependent on both a trypsin-sensitive component and cell
ular energy. Uptake in these cells was nonspecific and did not appear
to be due to receptor-mediated endocytosis. Therefore, because oligonu
cleotide cellular uptake in other cell types apparently involves an en
docytic mechanism, the primary mechanism of oligonucleotide internaliz
ation may be cell line dependent.