慢病毒载体介导RNAi稳定抑制VASH1表达对人脑胶质瘤细胞增殖、凋亡影响的体外研究
DOI: 10.3971/j.issn.1000-8578.2016.03.005
Keywords: VASH1,RNA干扰,慢病毒载体,胶质瘤,Effect of Down-regulating KLF8 Expression by siRNA on EMT in Nasopharyngeal Carcinoma Cells,Effect of Gene Silencing of RNF2 on Radiosensitivity of Esophageal Carcinoma Cells,Effect of RNA Interference Decreasing MSI1 Expression on Chemosensitivity of Human Glioma Cells,MDM2 siRNA Inhibits Angiogenesis by Reducing Vascular Endothelial Growth Factor Production in Breast Cancer Cells,Effect of siRNA-mediated notch1 Gene Silencing on Apoptosis and Radiosensitivity of Glioma Cell Line U87-EGFRvⅢ,Review on Radiation Dose Fractionation on High-grade Glioma Patients,LncRNA Colorectal Neoplasia Differentially Expressed(CRNDE) Regulated Cell Proliferation and Migration in Glioma,GM-CSF Enhances Effect of TMZ on High-grade Glioma Cells,Effects for Histone Deacetylase Inhibitors on Proliferation and MKK7 Expression in Glioma Cells,Relationship Between FoxQ1 and Epidermal Growth Factor Receptor Gene Expression in Colorectal Cancer,Effect of HSP70 and iNOS in Process of Hyperthermia Reducing Glioma Invasion,Construction and Screening of shRNA Expression Plasmids Targeting at cortactin Gene,Research Status and Development of Adherens Junction Associated Protein-1 in Gliomas,Effects of Vascular Endothelial Cells on Invasion of Glioma Cells via Notch Signal Pathway,Down-regulating UHRF1 Expression could Inhibit Progression of Hepatocellular Carcinoma
Abstract:
摘要 目的 探讨运用慢病毒载体介导的RNA干扰技术抑制血管生成抑制因子1（VASH1）的表达对人脑胶质瘤U-87MG细胞增殖和凋亡的影响。方法 应用pGCL-GFP质粒构建针对VASH1的慢病毒shRNA载体，转染入工具细胞293T，筛选出适合浓度的慢病毒转染人脑胶质瘤U-87MG细胞；通过RT-PCR和Western blot评价其对U-87MG细胞内VASH1表达的影响；用四甲基偶氮唑蓝（MTT）法观察U-87MG细胞增殖活性的改变；用流式细胞仪（FCM）检测U-87MG细胞凋亡的变化。结果 与转染阴性对照组和空白组相比，转染pGCL-GFP-VASH1慢病毒载体的U-87MG细胞VASH1的mRNA 蛋白表达水平明显降低（P<0.01）；细胞的增殖活性显著上调（P<0.01），细胞凋亡明显减少（P<0.01）。结论 VASH1 shRNA慢病毒载体可抑制VASH1在人脑胶质瘤U-87MG细胞中的表达，并增加肿瘤细胞的增殖活性，抑制细胞凋亡
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