Effect of Down-Regulating the CD59 by RNAi Lentivirus on the Expression of Acute T-lineage Leukemia Jurkat Cell Line.
10.7534/j.issn.1009-2137.2019.06.006
- Author:
Li-Ping WANG
1
;
Chang-Ming SUN
2
;
Zheng-Xiang HUA
1
;
Li-Na YAN
3
Author Information
1. Department of Blood Transfusion, The Affiliated Hospital of Chengde Medical College, Chengde 067000, Hebei Province, China.
2. Department of South District Laboratorial Examination, The Affiliated Hospital of Chengde Medical College, Chengde 067000, Hebei Province, China.
3. Department of Hematology, The Affiliated Hospital of Chengde Medical College, Chengde 067000, Hebei Province, China,E-mail：fysxk@sina.com.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
CD59 Antigens;
Cell Lineage;
Cell Proliferation;
Down-Regulation;
Humans;
Jurkat Cells;
Lentivirus;
Leukemia;
RNA Interference;
RNA, Small Interfering;
Transfection
- From:
Journal of Experimental Hematology
2019;27(6):1744-1748
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To analyze the effect of down-regulating the CD59 gene expression by RNAi lentivirus as vector on Jurkat cell line of acute T-lineage leukemia.
METHODS:The expression of CD59 in Jurkat cell line of acute T-line leukemia was induced to decrease by RNAi lentivirus as vector. The transfection of RNA lentivirus and the localization of CD59 molecule were analyzed by laser confocal technique. The relative expression of CD59 gene in blank control, negative control and RNAi lentivirus transfected group was detected by real-time fluorescence quantitative PCR, and the enzyme-linked immunosorbent assay was used to detect the expression of TNF-β and IL-3 in supernatants of cultured cells in 3 groups. The expression levels of apoptosis-related molecules including Caspase-3, Survivin, BCL-2 and BCL-2-associated X protein (BAX) were measured by Western blot.
RESULTS:The transfection efficiency for Jurkat cells was higher than 90%. CD59 was mainly located on the cell membrane. Compared with the blank control group and the negative control group, the expression level of CD59 mRNA and protein in the RNAi lentivirus transfected group significantly decreased (P<0.05). Compared with the blank control group and the negative control group, the expression of TNF-β and IL-3 in the RNAi lentivirus transfected group were significantly higher and lower (P<0.05) respectively. The expression levels of Survivin and BCL-2 in the RNAi lentivirus transfected group were significantly lower than those in the blank control group and the negative control group, while the expression levels of Caspase-3 and BAX in the RNAi lentivirus transfected group were significantly higher than those in the blank control group and the negative control group (P< 0.05).
CONCLUSION:The down-regulation of CD59 gene expression induced by RNAi lenti-virus can decrease the expression of proliferation and differentiation-promoting molecule such as IL-3 and increase the expression of TNF-related factor in Jurkat cell line of acute T-lineage leukemia, which also can increase the expression of apoptosis-related proteins such as Caspase-3 and BAX, and decrease the expression of anti-apoptosis-related proteins such as Survivin and BCL-2.
