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目的:研究ADAR1 shRNA对人胶质瘤细胞U87细胞增殖和凋亡的影响。方法:通过构建ADAR1-shRNA的干扰质粒,经脂质体法转染胶质瘤U87细胞系,通过荧光倒置显微镜观察转染效率,选择转染效率最高的细胞系。取转染48h细胞,采用RT-PCR和Western-blot分别检测ADAR1 mRNA及蛋白的表达,流式细胞仪检测其细胞凋亡率,MTT法检测细胞增殖情况。结果:①经ADAR1-shRNA转染48h后的转染效率最高,此时U87细胞系中ADAR1 mRNA及蛋白的表达均被显著抑制,较阴性对照组及空白组均明显降低(P<0.05)。②在转染ADAR1-shRNA后,细胞凋亡率为(28.14%±3.76%),明显高于阴性对照组(3.20%±1.57%)和空白组(2.80%±1.49%),细胞增殖率较阴性对照组及空白组明显下降(P<0.05)。结论:通过shRNA抑制ADAR1的表达能明显促进人胶质瘤细胞U87细胞的凋亡和抑制其增殖,ADAR1基因可能成为治疗治疗胶质瘤的新靶点。
Objective: To investigate the effect of ADAR1 shRNA on proliferation and apoptosis of human glioma U87 cells. METHODS: The ADRI1-shRNA interference plasmid was constructed and transfected into glioma U87 cell line by lipofectamine. The transfection efficiency was observed by fluorescence inverted microscope, and the cell line with the highest transfection efficiency was selected. The expression of ADAR1 mRNA and protein were detected by RT-PCR and Western-blot respectively. The apoptosis rate of ADAR1 was detected by flow cytometry. The proliferation of cells was detected by MTT assay. Results: ① The transfection efficiency was highest at 48h after transfection with ADAR1-shRNA. The expression of ADAR1 mRNA and protein in U87 cell line was significantly inhibited at this time, which was significantly lower than that in the negative control group and the blank group (P <0.05). ② After transfecting ADAR1-shRNA, the rate of apoptosis was (28.14% ± 3.76%) which was significantly higher than that of negative control group (3.20% ± 1.57%) and blank control group (2.80% ± 1.49% Negative control group and blank group decreased significantly (P <0.05). Conclusion: Inhibition of ADAR1 expression by shRNA can significantly promote the apoptosis of U87 cells and inhibit its proliferation. ADAR1 gene may be a new therapeutic target for gliomas.