Effect of NACK knockdown on the proliferation and apoptosis of T-cell acute lymphoblastic leukemia Jurkat cells
10.3760/cma.j.cn101070-20200404-00579
- VernacularTitle:NACK的下调对急性T淋巴细胞白血病Jurkat细胞增殖、凋亡的影响
- Author:
Linlin LI
1
;
Aimin LI
;
Jianyong WANG
;
Haiyan XING
;
Xiaoli WANG
;
Li WANG
;
Jianying LIU
Author Information
1. 青岛大学附属烟台毓璜顶医院儿内科，山东 烟台 264000
- Keywords:
Notch activation complex kinase;
Notch1 signaling pathway;
T-cell acute lymphoblastic leukemia
- From:
Chinese Journal of Applied Clinical Pediatrics
2021;36(15):1181-1184
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect and mechanism of NACK knockdown on the proliferation and apoptosis of T-cell acute lymphoblastic leukemia (T-ALL) Jurkat cells. Methods:Lentivirus transfection technology was used to transfect Jurkat cells and knock down NACK gene.Real time fluorescent quantitative PCR and Western blot were used to detect the silencing efficiency of NACK gene.CCK-8 method and flow cytometry were used to detect the effects of NACK knockdown on the proliferation and apoptosis of Jurkat cells.The expressions of protein related with Notch1 pathway, such as Hes1 and c-Myc, were detected by Western blot. Results:After NACK-shRNA was successfully transfected into Jurkat cells by lentiviral vector, the expression of NACK mRNA and protein was reduced signi-ficantly ( P<0.05). Compared with the negative control group and the blank control group, the CCK-8 method showed that the cell proliferation in the experimental group was significantly inhibited [The inhibition rates of cell proliferation in the experimental group, negative control group and blank control group were (37.27±4.48)%, (4.25±2.10)% and (2.43±1.40)%, respectively]( F=132.640, P<0.05), and the flow cytometry test showed that the apoptosis in the experimental group increased significantly [The apoptosis rates of experimental group, negative control group and blank control group were (26.38±3.03)%, (6.07±2.61)% and (3.40±1.98)%, respectively]( F=90.534, P<0.05). Western blot results confirmed that the expression of Notch1 pathway-related proteins Hes1 and c-Myc was down-regulated compared with the negative control group and the blank control group, and the difference was statistically significant ( P<0.05). Conclusions:Targeting silent NACK can down-regulate the expression of Notch1 pathway-related proteins, which leads to the inhibition of Jurkat cell proliferation and increased apoptosis, thereby exerting its anti-T-ALL effect.
