Volume 16 No 5 (2018)
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Effects of siRNA Targeting Interference of APE1 Gene on Proliferation and Apoptosis of Glioma Cells
Rui Wang , Bo Wei , Le Wang , Daliang Kong , Guozhang Hu , Chao Du
Abstract
This paper explored the application of RNA in cancer gene therapy. Second, the effect of the recombinant plasmid
APE1 siRNA on the expression of APE1 protein was observed. In addition, the proliferation of cells in apoptosis and
the radiosensitivity of human glioma U251 cells were also discussed. U251 cells were divided into control group,
negative group and APE1siRNA group for culturing. There were two holes in each group, and independent
experiments were repeated three times. The expression of APE1 protein in U251 cells was detected by Western
blot experiments for siRNA transfection. The siRNA cells were transfected, while another group of FAM fluorescent
labeled non-sense sequence siRNA was transfected to determine the transfection rate and extract the total protein
of the cell. BCA protein assay kit was used for determining the concentration. Meanwhile, 50g sample and SDSpolypropylene diamine gel were prepared. Coomassie blue staining was used to observe the protein
electrophoresis, after which the protein on the gel was transferred to the PVDF membrane and then developed in a
chemiluminescent detection system with specific antibodies. The β -action test was used as an internal controller,
while the optical density scanner was used to detect the gray value of the target strip. The relative expression of
target protein was equal to the target band gray value or the same sample reference value of gray. MTT method was
used to detect the cell growth curve experiments, while AO or EB method was used to detect the apoptosis rate of
glioma cells. SPSS17.0 was used for statistical analysis. The results show that the APE1 siRNA expression plasmid
was successfully introduced into human glioma cells by RNA interference, which can reduce the expression of APE1
in cells. It was confirmed that the reduction of APE1 protein could inhibit proliferation and promote the apoptosis
of glioma cells. Third, APE1 targeting interference may be an effective gene therapy strategy for human glioma,
which can provide an experimental basis for the clinical gene therapy of glioma.
Keywords
APE1 Targeting Interference, MTT Method, Apoptosis Rate
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