NEDD9在神经胶质瘤中的表达及基因沉默对人胶质瘤U251细胞增殖与侵袭的影响

The expression of NEDD9 in glioma and the effect of gene silencing on the proliferation and invasion of human glioma U251 cells

目的研究NEDD9在神经胶质瘤中的表达及NEDD9基因沉默对人胶质瘤U251细胞增殖与侵袭的影响。方法选取手术切除的脑胶质瘤组织36例及正常脑组织10例,采用qRT-PCR和Western blot法检测各组脑组织中NEDD9mRNA和蛋白表达情况,构建靶向NEDD9的shRNA重组慢病毒载体,将慢病毒载体转染U251细胞,应用qRT-PCR与Western blot方法检测NEDD9mRNA与蛋白表达,MTT方法检测转染后U251细胞活力的变化,Transwell实验检测细胞侵袭能力的变化。结果脑胶质瘤组织NEDD9的蛋白与mRNA表达水平显著高于正常脑组织。成功建立了稳定沉默NEDD9基因的U251细胞株,与空白对照组及阴性对照组比较,转染NEDD9-shRNA组U251细胞活力与侵袭能力显著降低(P<0.01)。结论 NEDD9可能是治疗胶质瘤U251细胞的潜在靶点。

Objective To investigate the expression of neural precursor cell expressed developmentally downregulated 9(NEDD9) in glioma and the effect of gene silencing on the proliferation and invasion of human glioma U251 cells. Methods The expression of NEDD9 in brain tissues was detected by qRT-PCR and Western blot in 36 cases of brain glioma tissues and 10 cases of normal brain tissues. The recombinant lentivirus vector targeting NEDD9 was constructed and transfected into U251 cells, and the expression of NEDD9 protein and mRNA was detected by qRT-PCR and Western blot respectively. MTT assay was used to detect the activity of U251 cells, and Transwell assay was used to detect the invasive ability of U251 cells. Results Compared with normal brain tissues, the expression level of NEDD9 protein and mRNA in glioma tissues was increased significantly. U251 cell line with stable silencing of NEDD9 gene was successfully established, the cell viability and invasive ability of U251 cells were significantly decreased in NEDD9-shRNA transfection group than in blank control group or negative control group(P<0.01). Conclusion NEDD9 might be a potential targer in the treatment of glioma U251 cells.