Effects of MLL-AF9 oncogene antisense oligonucleotide-mediated gene silencing on cell proliferation and apoptosis of THP-1 cells
10.3760/cma.j.issn.1009-9921.2008.04.001
- VernacularTitle:反义MLL-AF9基因对THP-1细胞增生和凋亡的影响
- Author:
Lei LI
;
Lingbo LIU
;
Li WANG
;
Ping ZOU
- Publication Type:Journal Article
- Keywords:
Oligoribonucleotieds,antisense;
MLL-AF9;
Leukemia monocytic,acute;
Cell proliferation;
Apoptosis
- From:
Journal of Leukemia & Lymphoma
2008;17(4):241-244
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the effects of antisense oligonucleofide(ASODN)fusion gene MLL-AF9 on cell proliferation and apoptosis of human acute monocytic leukemia cell line THP-1 expressing MLL-AF9.Methods ASODN and sense oligonucleotide(SODN)targeting MLL-AF9 were designed,constructed and transfected into THP-1 by lipofectmine.The level of MLL-AF9 mRNA expression was examined by reverse transcription polymerage chain reaction(RT-PCR)and the expression of MLL-AF9 protein was detected by Western blotting. Cell proliferation rate was analyzed by modified-MTT assay.Hoechst33258 staining Wag used to detect the apoptotic bodies in the cells.The change of apoptosis rate was detected by flow cytometry.Results The level of MLL-AF9 mRNA and protein expression after transfection was significantly decreased in ASODN-treated group in comparison with that in the control group,liposome group and SODN group(P＜0.01).Proliferation of the ASODN-treated cells was inhibited and apoptosis was evidently increased by(10.4±3.0)%,(22.8±2.5)%and(24.7±3.1)% for 0,24 and 48 h,respectively in a time-dependent manner (P＜0.01).Staining of the ASODN-treated cells with Hoechst 33258 showed the morphology characteristic of apoptosis.Conclusion Special ASODN targeting MLL-AF9 effectively inhibited MLL-AF9 expression in THP-1 cell,reduced cell proliferation and induced apoptosis.
