| 刘藻滨蒋晓帆张磊饶维杨悦凡戴舒惠陈涛
李三中罗鹏李娟费舟△.ADAR1 shRNA 对人胶质瘤U87 细胞增殖和凋亡的影响*[J].,2014,14(17):3232-3235 |
| ADAR1 shRNA 对人胶质瘤U87 细胞增殖和凋亡的影响* |
| Effects of shRNA-ADAR1 on the Proliferationand Apoptosis of U87 Glioma Cell* |
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| DOI： |
| 中文关键词: ADAR1 胶质瘤 增殖 凋亡 |
| 英文关键词: ADAR1 Glioma Proliferation Apoptosis |
| 基金项目:国家自然科学基金项目(30930093；81071034) |
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| 中文摘要: |
| 摘要目的：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。 |
| 英文摘要: |
| ABSTRACT Objecive：This studywas designedtoinvestigate the effects of transfectionwith small hairpinRNA(shRNA)-ADAR1 on
the proliferation and apoptosis of human glioma U87 cell line. Methods:The shRNA expression vector which expresses the specific
shRNA targeting ADAR1 mRNA (ADAR1-shRNA) or independent sequence (Negative-shRNA) was transfected into U87 cells, and then
the U87 cells with stable expression of shRNA were selected. The expressions of ADAR1 were detected by RT-PCR and Western
blotting. The cellular proliferation activity, and the apoptotic rate were determined by MTT assay and flow cytometry, respectively.
Results:Compared with the Negative-shRNA group and the non-transfection group, the mRNA and protein expression levels of ADAR1
in U87 cells with transfection of ADAR1-shRNA (interference group) was significantly suppressed and the cell proliferation was slowed
down after transfection for 48h (P＜0.05). The apoptotic rate in the interference group (28.14%± 3.76%) was significantly higher than
those in the Negative-shRNA group (3.20%± 1.57%) and the non-transfection group (2.80%± 1.49%) (P＜0.05). The proliferation ratio
of U87 cells significantly decreased after transfection of ADAR1 shRNA (P<0.05).Conclusion: ADAR1-shRNA transfection increases
the apoptotic rate, inhibits the proliferation of U87 glioma cells, which provides a potential target in gene therapy for glioma. |
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