Cationic lipids enhanced cellular uptake and activity of bcl-2 antisense oligodeoxynucleotide G3139 in HL-60 cells.
OBJECTIVE
To explore the effect of cationic lipid 1,3-di-oleoyloxy-2-(6-carboxy-spermyl)-propylamid (DOSPER) on cellular uptake and activity of bcl-2 antisense oligodeoxynucleotide G3139 in HL-60 cells.


METHODS
The cell-associated mean fluorescence intensity and the percentage of cells with positive staining for Bcl-2 were measured by flow cytometry. The subcellular distribution of fluorescein isothiocyanate (FITC)-labeled G3139 was observed by fluorescence microscope and the bcl-2 mRNA level was detected by reverse transcription polymerase chain reaction (RT-PCR).


RESULTS
(1) DOSPER increased cellular uptake of G3139 into HL-60 cells greatly. When DOSPER/G3139 (microg : microg) was 2 : 1, the uptake of G3139 reached top after treatment for 2 h and increased about 20 times compared with application of G3139 alone. In the presence of DOSPER, G3139 was localized in nucleus and cytosol with a bright spotted fluorescence staining. However, G3139 was localized in cytoplasm with faint fluorescence in the absence of DOSPER. (2) Cell-associated G3139 could be effluxed out of cells. After treated with G3139 in the presence of DOSPER for 4 h, the cell-associated G3139 could be fitted by C(t)=68.2e(-0.60) t+31.8e(-0.02) t (% of initial value), with a half-life of approximately 1.1 h. In the absence of DOSPER, the cell-associated G3139 could be fitted by C(t)=64.8e(-2.27) t+35.2e(-0.04) t, with a half-life of about 18 min. (3) In the presence of DOSPER, G3139 1.0 micromol/L specially reduced bcl-2 mRNA level, and Bcl-2 protein decreased from 97 % +/- 4 % to 70.6 % +/- 2.1 %.


CONCLUSIONS
DOSPER enhanced the activity of G3139 and it might be attributed to increase of the cellular uptake and change of the subcellular distribution of G3139.
Zhu
Y G
YG
Fujian Institute of Hematology, Union Hospital, Fujian Medical University, Fuzhou 350001, China. yuanguizhu@163.net
Zhuo
G S
GS
Chen
Z Z
ZZ
Chen
X C
XC
D001692:Biological Transport / Q000187:drug effects; D027682:Cation Transport Proteins; D005229:Fatty Acids, Monounsaturated / Q000494:pharmacology*; D018922:HL-60 Cells / Q000378:metabolism; D006801:Humans; D008055:Lipids / Q000494:pharmacology; D020319:Oligodeoxyribonucleotides, Antisense / Q000378:metabolism*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000037:antagonists & inhibitors / Q000235:genetics; D013873:Thionucleotides / Q000378:metabolism*
C118665:1,3-dioleoyloxy-2-(6-carboxyspermyl)propylamide; D027682:Cation Transport Proteins; D005229:Fatty Acids, Monounsaturated; D008055:Lipids; D020319:Oligodeoxyribonucleotides, Antisense; D019253:Proto-Oncogene Proteins c-bcl-2; D013873:Thionucleotides; C408162:oblimersen