[Change of Bcl-2, Bax proteins and mitochondrial membrane protein on nitric oxide induced apoptosis in HL-60 cells].
To investigate the possible mechanisms of nitric oxide (NO)-induced apoptosis in leukemia cell line HL-60, HL-60 cells in vitro were incubated with sodium nitroprusside (SNP), the in situ cell apoptosis quantitatively was assayed by TdT-mediated dUTP nick end labeling (TUNEL), the cell cycle DNA and proteins expression of Bcl-2, Bax, mitochondrial membrane protein (APO2.7) were analyzed by flow cytometry. The results showed that SNP induced HL-60 cell apoptosis in a dosage- and time-dependent manner. After exposure to SNP at the concentration of 1.0 mmol/L for 48 hours, the percentage of apoptosis HL-60 was (42.2 +/- 3.5)% for subG1 and (52.5 +/- 7.6)% for TUNEL respectively, and they are significantly higher than those in control and potassium ferricyanide (PFC) groups as same concentration. During the apoptosis process, it showed a decrease of Bcl-2 protein and an increase of Bax protein and mitochondrial membrane protein in HL-60 cell, proteins of Bcl-2, Bax and mitochondrial membrane were expressed in a dosage- and time-dependent manner too. In conclusion, during the process of SNP induced apoptosis in HL-60 cell, the expression of mitochondrial membrane protein was increased, Bcl-2 and Bax proteins may be important regulators.
Zhong
Yong-Lie
YL
Central Laboratory, Zhejiang Provincial People Hospital, Hangzhou 310014, China. zyl63@mail.hz.zj.cn
Qiu
Lian-Nu
LN
Lin
Hui-Jun
HJ
Wang
Wen-Song
WS
Wu
Jian-Guo
JG
Liu
Jian-Dong
JD
D017209:Apoptosis / Q000187:drug effects*; D018922:HL-60 Cells; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis*; D024101:Mitochondrial Proteins / Q000032:analysis*; D020030:Nitric Oxide Donors / Q000494:pharmacology*; D009599:Nitroprusside / Q000494:pharmacology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000032:analysis*; D051028:bcl-2-Associated X Protein
C494826:BAX protein, human; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D020030:Nitric Oxide Donors; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D009599:Nitroprusside