Fluacrypyrim induced apoptosis in acute myeloid leukemia cell lines
10.13220/j.cnki.jipr.2018.01.009
- VernacularTitle:嘧螨酯诱导急性髓系白血病细胞凋亡
- Author:
Lu ZHANG
1
;
Xing SHEN
;
Fang-Min WANG
;
Shuang XING
;
Guo-Lin XIONG
;
Hong-Ling OU
;
Xin-Ru WANG
;
Zu-Yin YU
Author Information
1. 121001锦州,锦州医科大学研究生院;100850北京,军事科学院军事医学研究院辐射医学研究所;100088北京,解放军火箭军总医院检验科
- Keywords:
fluacrypyrim;
acute myeloid leukemia(AML);
apoptosis
- From:
Journal of International Pharmaceutical Research
2018;45(1):51-56
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of fluacrypyrim on the induction of apoptosis in human acute myeloid leuke-mia(AML)cell lines,NB4,THP-1 and HL-60,and explore the related mechanisms.Methods Trypan blue dye exclusion assay was used to estimate the growth of NB4,THP-1,and HL-60 cells after treatment with various concentrations of fluacrypyrim(1.25, 2.5,5 and 7.5 μmol/L)for 72 h.Cell apoptosis was evaluated by AnnexinⅤ-FITC/PI double stainning for the NB4 and THP-1 cells treated with fluacrypyrim(1.25,2.5 and 5 μmol/L)for 48 h as well as the HL-60 cells treated with fluacrypyrim(2.5,5 and 7.5 μmol/L) for 72 h.Western blotting was used to examine the protein expression of apoptotic regulators Bax,Mcl-1 and Caspase 3 in the NB4 cells treated with fluacrypyrim(1.25,2.5 and 5 μmol/L)for 24 h.Then,NB4 Cells were pretreated with Caspases inhibitor benzyloxy-carbonyl-Val-Ala-Asp-fluoromethylketone(Z-VAD-FMK)and exposed to fluacrypyrim at 2.5 μmol/L for 24 h,which was then evaluat-ed for the apoptosis using AnnexinⅤ-FITC/PI double stainning.Western blotting was used to examine the expression of the phosphory-lated and total proteins of mitogen-activated protein kinase(MAPK)signaling molecules,ERK,JNK and P38,in the NB4 cells treat-ed with fluacrypyrim(1.25,2.5 and 5 μmol/L)for 24 h. NB4 Cells were pretreated with ERK inhibitor U0126,JNK inhibitor SP600125,or P38 inhibitor SB203580 for 1 h and then exposed to fluacrypyrim at 2.5 μmol/L for 24 h,which was then analyzed for the apoptosis by the AnnexinⅤ-FITC/PI double stainning.Results The proliferation of NB4,THP-1 and HL-60 cells was inhibited by the treatment with fluacrypyrim(2.5,5 and 7.5 μmol/L)for 72 h.The apoptosis induced in the NB4 and THP-1 cells by the fluacry-pyrim treatment at 5 μmol/L for 48 h and in the HL-60 cells by the fluacrypyrim treatment at 7.5 μmol/L for 72 h were significant as compared with the control group(P<0.01).Mechanically,fluacrypyrim at the concentrations of 2.5 and 5 μmol/L effectively up-regu-lated the expression of Bax(P<0.01 and P<0.05)for the 2.5 and 5 μmol/L,respectively,down-regulated the expression of Mcl-1 (P<0.01)and activated Caspase 3(P<0.01)in the NB4 cells when compared with the control group(P<0.01).The pretreatment of the NB4 cells with Z-VAD-FMK blocked the apoptosis induced by fluacrypyrim.Furthermore,the fluacrypyrim(2.5 and 5 μmol/L) treatment increased the ERK,JNK and P38 phosphorylation(P<0.01),while the pretreatment of the NB4 cells with U0126 signifi-cantly inhibited the the fluacrypyrim-induced apoptosis(P<0.01),as compared with the control group.Conclusion Fluacrypyrim effectively inhibits the cell proliferation and induces caspase-dependent cell apoptosis in AML cells.Activation of ERK/MAPK signal-ing pathway might play an important role in the action of fluacrypyrim.
