[The effects of JARID1B siRNA on proliferation and apoptosis in HL-60 cell].
OBJECTIVE
To study the effect of small interfering RNA(siRNA) targeting JARID1B gene on the proliferation and apoptosis in HL-60 acute promyelocytic leukemia cell line, and to explore its mechanisms.


METHODS
The JARID1B siRNA was transfected into HL-60 cells using Lipofectamine(TM) 2000(Lipo) vector. The proliferation inhibition by siRNA targeting JARID1B was detected by MTT, cells apoptosis by flow cytometry, the mRNA expression of JARID1B by RT-PCR, the protein expression of JARID1B, Bcl-2, procaspase-9, procaspase-3, c-myc and P27 and histone methylated H3K4 by Western blot.


RESULTS
siRNA targeting JARID1B upregulated histone methylated H3K4 level, inhibited the proliferation of HL-60 cells, and induced the cells apoptosis. After transfection of siRNA targeting JARID1B at 0, 30, 60, 120 nmmol/L for 24 hours, the apoptotic rate were (11.0 ± 3.6)%, (35.2 ± 5.1)%, (52.7 ± 3.8)%, and (62.0 ± 5.7)% respectively (F = 70.27, P < 0.01). The protein expression of P27 was upregulated, and Bcl-2, procaspase-9, procaspase-3, c-myc was down regulated.


CONCLUSIONS
JARID1B siRNA upregulates histone methylated H3K4. It reduces HL-60 cells proliferation and apoptosis by up regulating the p27 expression and down regulating the Bcl-2, procaspase-9, procaspase-3, c-myc expression. It might be a new therapeutic targeting for human leukemia.
Department of Hematology, Fujian Medical University, Zhangzhou, China.
Xu-dong
XD
Ma
Hui-dan
HD
Han
Yi-qun
YQ
Huang
Yong
Y
Zou
D017209:Apoptosis; D053148:Caspase 3 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D049109:Cell Proliferation; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000378:metabolism; D015973:Gene Expression Regulation, Leukemic; D018390:Gene Targeting; D018922:HL-60 Cells; D006657:Histones / Q000378:metabolism; D006801:Humans; D056484:Jumonji Domain-Containing Histone Demethylases / Q000235:genetics*; D007938:Leukemia / Q000235:genetics; D008745:Methylation; D009687:Nuclear Proteins / Q000235:genetics*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering; D012097:Repressor Proteins / Q000235:genetics*
C436291:CDKN1B protein, human; D006657:Histones; C489427:MYC protein, human; D009687:Nuclear Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D016271:Proto-Oncogene Proteins c-myc; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012097:Repressor Proteins; D050760:Cyclin-Dependent Kinase Inhibitor p27; D056484:Jumonji Domain-Containing Histone Demethylases; C119611:KDM5B protein, human; D053148:Caspase 3; D053453:Caspase 9