shRNA敲低ILK后对人脑胶质瘤细胞U251增殖迁移及侵袭影响.docVIP

shRNA敲低ILK后对人脑胶质瘤细胞U251增殖迁移及侵袭影响 　　【摘要】 目的：观察应用shRNA敲低人脑胶质瘤U251细胞系ILK基因表达后，对其在增殖、迁移、侵袭的影响。方法：将特异性的shRNA表达载体质粒ILK-PGFP-V-RS转染人脑胶质瘤细胞系U251（U251-ILK），同时构建空质粒组（U251-NC）稳定转染细胞株。运用RT-PCR及蛋白质免疫印迹（Western blot）测定各组细胞株的ILK、cyclin D1、E-cadherin、Snail mRNA及其蛋白表达水平，应用Transwell及预铺 Matrigel的小室检测转染后相应细胞株的迁移和侵袭能力，同时使用CCK8法检测细胞株增殖能力。结果：顺利构建成具有稳定敲低ILK表达水平的U251-si细胞株。U251-si细胞已证实ILK无论在mRNA水平还是其蛋白质水平均较U251组、U251-N显著降低（P0.05）。结论：应用shRNA敲低胶质瘤细胞系U251中ILK基因后，抑制了胶质瘤细胞的增殖迁移及侵袭能力，为临床治疗提供新的作用靶点。 　　【关键词】 神经胶质瘤； shRNA； 整合素连激酶； E-钙粘蛋白 　　中图分类号 R739.41 文献标识码 A 文章编号 1674-6805（2015）9-0012-04 　　The Effect of Knockdown ILK by shRNA on the Proliferation，Migration and Invasion of Human Glioma Cell U251/WANG Guang-yi，ZHENG Ke-bin，LI Chun-hui.//Chinese and Foreign Medical Research，2015，13（9）：12-15 　　【Abstract】 Objective：To investigate the change of tumorigenesis，proliferation，migration and invasion of human brain cells U251 after knockdown ILK by shRNA interference.Method：U251 were transfected with the specific ShRNA（ILK-PGFP-V-RS） vector（U251-si），and empty vector were constructed as a negative control（U251-N）.Stable transfected cells were established.The expression of ILK Cyclin D1 E-cadherin Snail mRNA and its protein among the stable transfected cells were detected by RT-PCR，Western blot，respectively.Cell migration and invasion ability were detected by Transwell assay（Matrigel matrix per well were used in vasion assay）.Result：The stable transfected cell lines U251，U251-si，U251-N were constructed.Conpared to the control cell，the anility of proliferation，migration，invasion were markedly inhibited in the cells（P0.05）.Conclusion：The inproliferation，migration and invasion are inhibited by knockdown ILK through shRNA in glioma cell U251，suggesting that loss of ILK may play a significant role in gliomagenesis. 　　【Key words】 Glioma； shRNA； ILK； E-cadherin 　　First-author’s address：Hebei University，Baoding 071000，China 　　doi：10.14033/j.cnki.cfmr.2015.09.005 　　神经胶质细胞瘤是目前最常见的原发性中枢神经系统肿瘤，其多起源于神经上皮组织，并且其确诊较