Effect of Regulating A20 Expression on NF-κB Expression and Biological Characteristics of Jurkat Cells.
10.19746/j.cnki.issn.1009-2137.2020.04.010
- Author:
Zhe WANG
1
;
Shi-Shan XIAO
1
;
Qian DING
1
;
Qing WANG
1
;
Sheng-Nan ZHOU
1
;
Zhe LI
1
;
Hong-Qian ZHU
2
Author Information
1. Department of Hematology, Guizhou Provincial People's Hospital, Guiyang 550002, Guizhou Province, China.
2. Department of Hematology, Guizhou Provincial People's Hospital, Guiyang 550002, Guizhou Province, China,E-mail: zhuhongqian@126.com.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
Cell Proliferation;
Humans;
Jurkat Cells;
NF-kappa B;
RNA, Small Interfering;
Transfection;
Tumor Necrosis Factor alpha-Induced Protein 3
- From:
Journal of Experimental Hematology
2020;28(4):1144-1151
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To explore the effect of regulating A20 expression on NF-κB and biological characteristics of Jurkat cells with glucocorticoid (GC) resistance.
METHODS:CCRF CEM and Jurkat cells were treated with dexamethasone (DEX) at concentrations of 100、10、1、0.1、0.01 and 0.001 μmol/L, and cultured for 24、48 and 72 h. The proliferation inhibition rate of Jurkat cell was detected by CCK-8. A20 plasmid was constructed, A20-siRNA was designed and synthesized, and transfected into Jurkat cells by liposome. CCK-8 was used to detect the proliferation rates of Jurkat cells in different concentrations of DEX group, DEX combined with A20 plasmid group and A20-siRNA group. The mRNA expression level of NF-κB was detected by RT-qPCR, the protein expression level of NF-κB was detected by Western blot, and the apoptosis of Jurkat cells was examined by flow cytometry.
RESULTS:The inhibitory effects of DEX at different concentrations on the growth of CCRF CEM cells were time-dependent (r=0.984, P＜0.05) and concentration-dependent (r=0.966, P＜0.05). At the point of 24 hour, the IC approached 1 μmol/L in CCRF CEM cells. Great large differences began to appear between 1 and 10 μmol/L, the proliferation rate of Jurkat cells treated with 1 μmol/L DEX did not show a significant change. Therefore, 1 μmol/L was selected as control group. The cell proliferation rate of A20 plasmid transfection combined with different concentrations of DEX group was lower than that of DEX group and A20-siRNA combined with DEX group. After transfection of A20 plasmid, the expression level of NF-κB was significantly lower than that of control group (P＜0.05), and the apoptotic rate was significantly higher than that of control group (P＜0.05). After transfection of Jurkat cells with A20-siRNA, the expression level of NF-κB was significantly higher than that of control group (P＜0.05). The apoptotic rate of cells in A20-siRNA group was not significantly changed (P＞0.05).
CONCLUSION:Jurkat cells are resistant to DEX. A20 overexpression combined with DEX can increase sensitivity of Jurkat cells with GC resistance and decrease the proliferation rate of Jurkat cells, down-regulate the expression level of NF-κB and promote the apoptosis of Jurkat cells.
