[Inhibitory effect of RNA interference on chronic myeloid leukemia bcr/abl oncogene expression].
OBJECTIVE
To investigate the inhibitory effect of RNA interference on chronic myeloid leukemia (CML) bcr/abl oncogene expression.


METHODS
The small interference RNAs (siRNAs) were synthesized in vitro. K562 cells stably expressing bcr/abl gene were transfected with the siRNA by electroporation, both the non-transfected cells and non-specific siRNAs transfected cells were taken as controls. The enhanced green fluorescent protein (EGFP) plasmid was used as positive control and the transfection efficiency was detected by flow cytometry. Inhibitory effect of siRNAs was demonstrated by real-time quantitative RT-PCR and Western blots. Cell proliferation was measured by MTT assay and apoptosis by Annexin V-FITC assay.


RESULTS
The transfection efficiency was about 70%. The synthesized siRNAs inhibited CML bcr/abl oncogene expression at both mRNA and protein levels. siRNAs could inhibit K562 cell proliferation to 47% and 56% at 24 h and 48 h after transfection, respectively, and induce cell apoptosis from 1.00% in control group to 15.05% and 19.4% at 24 h and 48 h respectively.


CONCLUSIONS
At the cell level, inhibition of CML bcr/abl oncogene expression by chemically synthesized siRNAs provides the new method for anti-leukemia study.
Ma
Xiao-xia
XX
Department of Hematology, Shanghai First People's Hospital, Shanghai 200080, China.
Wang
Chun
C
Wei
Ju
J
Qin
You-wen
YW
Yan
Shi-ke
SK
Gao
Yan-rong
YR
Cai
Qi
Q
D017209:Apoptosis / Q000235:genetics; D049109:Cell Proliferation; D016044:Fusion Proteins, bcr-abl / Q000235:genetics*; D006801:Humans; D020014:K562 Cells; D034741:RNA, Small Interfering; D014162:Transfection
D034741:RNA, Small Interfering; D016044:Fusion Proteins, bcr-abl