靶向抑制WISP1基因增强脑胶质瘤放射敏感性研究

Targeted inhibition of wisp1 enhanced radiosensitivity in glioma cells

目的 观察WISP基因与人脑胶质瘤细胞放射敏感性及迁移能力的相关性.方法 应用Western印迹法检测WISP1蛋白在人脑胶质瘤细胞中的表达量.利用慢病毒介导的RNA干扰技术沉默M059K细胞中WISP1基因.应用克隆形成实验和划痕实验观察细胞放射敏感性和细胞迁移能力的变化.结果 M059K细胞WISP1蛋白表达量高于M059J细胞,M059K细胞较M059J细胞放射敏感性低、细胞迁移能力强;慢病毒载体pGLV3/H1/GFP＋ puro vector-WISP1转染M059K后,WISP1蛋白表达明显下降,沉默WISP1基因后M059K细胞放射敏感性增强,迁移能力降低.结论 WISP1基因高表达的细胞系M059K较WISP1基因低表达的细胞系M059J放射敏感性低、迁移能力强.沉默WISP1基因后可以增强脑胶质瘤的放射敏感性,抑制其迁移能力.

Objective To explore the effects of differencial expression of WISP1 upon the radiosenstivity and migration in glioma cell lines M059K and M059J,as well as WISP1 knockdown cell line M059K-WISP1-.Methods The expression of WISP1 in different cell lines was detected by Western blot assay.The lentivirus mediated-RNA interference technology was employed to knockdown the endogenous expression of gene WISP1 in human glioma cells （M059K）.A stable cell line was established by infecting M059K cells with the lentivirus particles,the transfection efficiency was examined by fluorescence microscopy.The radiosensitivity and migration ability of cells were assessed by Colony-forming and Scratch Wound Assays respectively.Results WISP1 was highly expressed in M059K cells compared with M059J cells.M059K cells were less radiosensitive than M059J cells,and had a better ability of migration.M059K cells were transfected into by the lentiviral vector pGLV3/H1/GFP ＋ puro vector-WISP1.And M059KWISP1-cell line with stable WISP1 downregulation was established successfully.Western blot demonstrated significantly downregulated WISP1 expression in M059K-WISP1-cells,which showed obviously improved radiosensitivity in Colony-forming assay and suppressed migration ability in Scratch Wound Assay.Colonyforming assay based on the muti-target/single-hit model indicated that radiosensitivity of M059K-WISP1-was significantly decreased.Conclusion Compared with WISP1 low expression cell line M059J,WISP1 high expression cell line M059K was less radiosensitive and had a better ability of migration.We have successfully established a glioma cell line M059K-WISP1-stably downregulating WISP1,which shows a better radiosensitivity and a lowered migration activity in vitro.This may provide a novel therapeutic target for the treatment of human glioma.