敲低IKKε抑制人胶质瘤细胞U251增殖和侵袭的体外研究  

Knock-down of IKKε inhibits proliferation and invasion of U251 glioma cells in vitro

作　　者：李会兵[1] 张安玲[1] 王坤[1] 王广秀[1] 康春生[1] 黄强[1] 

机构地区：[1]天津医科大学总医院神经病学研究所神经肿瘤室,天津300052

出　　处：《中华神经医学杂志》2012年第2期121-125,共5页Chinese Journal of Neuromedicine

摘　　要：目的 探究敲低IKKε表达对人胶质瘤细胞系U251生物学特征的影响。方法采用脂质体介导的IKK8小干扰RNA（IKKesiRNA）转染U251细胞，同时设转染无义序列组和对照组，RT-PCR鉴定转染后IKKEmRNA的表达；MTT法检测转染后细胞的增殖能力：流式细胞术检测细胞周期分布的改变；Transwell实验检测细胞侵袭能力的变化；Westernblotting法检测核增殖抗原（PCNA）、细胞周期素Dl（CyclinD1）、基质金属蛋白酶．9（MMP9）和NF．KBP65的表达。结果与对照组和无义序列组比较，转染IKK／εsiRNA组细胞IKKemRNA的表达和细胞增殖率降低．G0／G1期细胞比例增高，穿膜细胞数明显减少，差异均有统计学意义（P〈0．05）。Westernblotting检测结果显示IKKεsiRNA组细胞PCNA、MMP9、CyclinD1的表达较对照组和无义序列组明显下调．而且NF-KB组分P65从胞浆向胞核的转座减少。结论IKKε在胶质瘤细胞增殖和侵袭过程中发挥重要作用，有望成为人脑胶质瘤基因治疗的侯选靶点。Objective To investigate the effect of IKKe knock-down on the biological characteristics of U251 glioblastoma cells. Methods IKKe small interfering RNA （LKKε siRNA） mediated by lipofectamine were transfected into U251 cells while cells transfected with scrambled siRNA and control cells were prepared. RT-PCR was employed to detect expressions oflKKe in the transfected cells. The cell proliferation was determined by MTT assay. Flow cytometry was used to monitor changes in cell cycle. Cell invasion was evaluated by Transwell assay. Moreover, the proliferating cell nuclear antigen （PCNA）, cyclin D1 and matrix metalloproteinase-9 （MMP9） that regulated proliferation, invasion and cycle progression of the transfected cells and the changes of NF-KB after transfection were examined by Western blotting. Results RT-PCR revealed that the proliferation of U251 cells was inhibited, the cell cycle was arrested in G0/G1 phase and the invasive activity was attenuated in cells transfected with IKKε siRNA, with significant differences as compared with the ceils transfected with scrambled siRNA and control cells （P〈0.05）. The expressions ofPCNA, MMP9 and cyclin D1 were down-regulated in the IKKε knock-down cells, as compared with the other 2 groups. In addtion, transposition of NF-KB reduced from the cytoplasm to the nucleus after transfection. Conclusion As IKKε plays a vital role in proliferation and invasion of glioma cells, it may serve as a potential target of gene therapy for glioma.

关 键 词：IKKε 神经胶质瘤 细胞增殖 侵袭性 细胞周期 

分 类 号：R739.4[医药卫生—肿瘤]