核因子-κB抑制物激酶ε促进人脑胶质母细胞瘤恶性进展的实验研究

Inhibitor of Nuclear Factor Kappa B Kinase Subunit Epsilon on the Malignant Progression of Human Glioblastoma

目的探讨核因子-κB抑制物激酶ε(IKKε)通过IKKε-Yes相关蛋白1(Yap1)-miR-Let-7b/i途径促进人脑胶质母细胞瘤恶性进展的具体作用机制。方法构建IKKε短发夹核糖核酸(shRNA)慢病毒及YAP1小干扰RNA(siRNA)。以IKKεshRNA慢病毒转染U87-MG细胞,下调细胞中IKKε的表达水平。通过RT-PCR及Western blot检测细胞中IKKε、YAP1及miR-Let-7b/i的表达;通过Transwell法检测细胞迁移和侵袭能力的变化。以YAP1 siRNA转染U87-MG细胞株,下调细胞中YAP1的表达水平。通过RT-PCR及Western blot检测细胞中YAP1及miR-Let-7b/i的表达。结果与对照组和无义序列组相比较,转染IKKεshRNA组IKKε蛋白及mRNA水平明显降低;YAP1蛋白水平明显降低;miR-Let-7b/i水平明显升高;穿过小室细胞数明显降低(P均<0.05)。与对照组和无义序列组相比较,转染YAP1 siRNA组YAP1蛋白及mRNA水平明显降低;miR-Let-7b/i水平明显升高(P均<0.05)。结论 IKKε可以通过IKKε-YAP1-miR-Let-7b/i途径促进人脑胶质母细胞瘤恶性进展,有望为人胶质母细胞瘤的治疗提供新的治疗策略。

Objective To investigate the effect of inhibitor of nuclear factor kappa B kinase subunit epsilon(IKKε)on the malignant progression of human glioblastoma by IKKε-YAP1-miR-Let-7b/i pathway.Methods IKKεshort hairpin ribonucleic acid(shRNA)lentivirus and YAP1 small interfere RNA(siRNA)were constructed.U87-MG cell lines were transfected with IKKεshRNA lentivirus to regulate the expression of IKKεin cells.The expressions of IKKε,YAP1 and miR-Let-7b/i were detected by RT-PCR and Western blot.The changes of cell migration and invasion were observed by Transwell assay.YAP1 siRNA were transfected into U87-MG cell lines to regulate the expression of YAP1 in the cells.The expressions of YAP1 and miR-Let-7b/i were detected by RT-PCR and Western blot.Results Compared with the control group and the nonsense sequence group,the levels of IKKεprotein and mRNA were significantly decreased in the IKKεshRNA lentivirus group(P<0.05),and the level of YAP1 protein was significantly decreased(P<0.05);the levels of miR-Let-7b/i were significantly increased(P<0.05);the number of cells passing through the transwell chamber was decreased in the IKKεshRNA lentivirus group(P<0.05).Compared with the control group and the nonsense sequence group,the levels of YAP1 siRNA and YAP1 protein and mRNA were significantly decreased(P<0.05),while the levels of miR-Let-7b/i in the YAP1 siRNA lentivirus group were significantly increased(P<0.05).Conclusion IKKεcan promote the malignant progression of human glioblastoma by IKKε-YAP1-miR-Let-7b/i pathway,and it is expected to provide a new therapeutic strategy for the treatment of glioblastoma.