[Effects of anti-PML-RAR alpha antisense on cell morphology and expression of PML-RAR alpha mRNA and protein of NB4 cells].
OBJECTIVE
To investigate the effects of anti-PML-RAR alpha antisense (FUAS) on cell morphology, expression of PML-RAR alpha mRNA and PML-RAR alpha/PML protein localization of NB4 cells.


METHODS
PML-RAR alpha mRNA expression was assayed by RT-PCR and PML-RAR alpha/PML protein localization by immuno-fluorescence.


RESULTS
NB4 cells were partially differentiated after 5 days of FUAS treatment and typical apoptosis was found after 7 days incubation with FUAS. The expression of PML-RAR alpha mRNA at 24 h was already down regulated in FUAS-treated cells. After 24 h, 72 h and 120 h incubation with FUAS, PML-RAR alpha mRNA showed 52.0%, 68.7% and 23.4% reductions, respectively, as compared with that of control. Immuno-fluorescence analysis with anti-PML monoclonal antibody showed disappearance of microgranules, residual granules becoming larger discrete dots at 24 h of FUAS treatment and almost disappearence of dots at 120 h.


CONCLUSIONS
FUAS specifically blocks the expression and translation of PML-RAR alpha gene, makes the production of PML-RAR alpha fusion protein decrease or disappear and prompts cell differentiation.
Chen
Y
Y
Shanghai Institute of Hematology, Renji Hospital, Shanghai Second Medical University, Shanghai 200001.
Miao
J
J
Zhu
X
X
D002455:Cell Division / Q000187:drug effects; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000473:pathology*; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*; D020319:Oligodeoxyribonucleotides, Antisense / Q000096:biosynthesis / Q000494:pharmacology*; D015514:Oncogene Proteins, Fusion / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D014407:Tumor Cells, Cultured
D009363:Neoplasm Proteins; D020319:Oligodeoxyribonucleotides, Antisense; D015514:Oncogene Proteins, Fusion; D012333:RNA, Messenger; C093476:promyelocytic leukemia-retinoic acid receptor alpha fusion oncoprotein