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.