[Design and synthesis of anti-PML-RAR alpha antisense and its effects on the growth and apoptosis of NB4 cells].
OBJECTIVE
To synthesize antisense oligodeoxynucleotides (AS) targeting the fusion point region (FUAS) and the start codon region (STAS) of the long type PML-RAR alpha mRNA and investigate its stability, specificity and effects on the growth and apoptosis of NB4 cells.


METHODS
NB4 cell apoptosis was assayed by flow cytometry, cell-fluorescence and DNA gel electrophoresis. Trypan blue exclusion was used for cell counts, and polyacrylamide gel electrophoresis for oligodeoxynucleotides.


RESULTS
The synthesized 18 bp phosphorothioate oligodeoxynucleotides was stable, resistant to nuclease, and no unspecific effects. Both STAS and FUAS could inhibit the NB4 cell growth in a dose-dependent manner. Low concentration (20 micrograms/ml) of STAS or FUAS resulted in growth inhibition of 0%-11.3%, and the highest inhibition was found at concentration of 80 micrograms/ml (inhibition rate 50.0%-67.7%). Cell DNA content analyzed by flow cytometry showed the typical profile of apoptotic cells at 7th day, 9th day of treatment with FUAS. Percentages of apoptotic cells in FUAS-treated cells was 9.3%, 24.5%, 41.0% and 34.2% after 3, 5, 7 and 9 days of FUAS treatment, respectively.


CONCLUSIONS
STAS and FUAS successfully synthesized and both of them could inhibit the growth and induce the cell apoptosis of NB4 cells.
Chen
Y
Y
Shanghai Institute of Hematology, Laboratory of Leukemia Research, Renji Hospital, Shanghai Second Medical University, Shanghai 200001.
Miao
J
J
Zhu
X
X
D017209:Apoptosis / Q000187:drug effects*; D002455:Cell Division / Q000187:drug effects; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000473:pathology*; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000494:pharmacology*; D020319:Oligodeoxyribonucleotides, Antisense / Q000096:biosynthesis / Q000494:pharmacology*; D015514:Oncogene Proteins, Fusion / Q000096:biosynthesis / Q000494:pharmacology*; D014407:Tumor Cells, Cultured / Q000187:drug effects
D009363:Neoplasm Proteins; D020319:Oligodeoxyribonucleotides, Antisense; D015514:Oncogene Proteins, Fusion; C093476:promyelocytic leukemia-retinoic acid receptor alpha fusion oncoprotein