摘要
目的探讨多亮氨酸重复区免疫球蛋白样蛋白1(LRIG1)对人脑胶质瘤细胞增殖的影响。方法分别构建携带LRIG1、LRIG1小干扰RNA、Bcl2和胶质细胞源性生长因子(BDMF)的质粒,将它们分别转染到人脑胶质瘤细胞系U251细胞内,应用细胞计数试剂盒8法检测细胞活性的变化,蛋白印迹法检测LRIG1、Bcl2、拓扑异构酶Ⅱ(topoⅡ)、GDNF的表达变化。结果成功构建携带LRIG1、LRIG1小干扰RNA、Bcl2、GDNF的质粒,并成功转入U251细胞株。过表达LRIG1显著抑制U251细胞增殖。蛋白印迹法结果显示LRIG1的表达和Bcl2、topoⅡ和GDNF的表达呈显著负相关。结论LRIG1通过负性调节Bcl2、topoⅡ和GDNF的表达抑制人脑胶质瘤细胞系U251细胞的增殖。
ObjectiveTo investigate the effect of leucine-rich repeats and immunoglobulin-like domains protein 1（LRIG1）on cell proliferation of glioma cell line U251.MethodsThe plasmids with LRIG1,LRIG1 siRNA,Bcl-2 or glial cell line-derived neurotrophic factor（GDNF）were constructed and then transfected into the U251 cells,respectively.The cell activities were detected by cell counting kit-8（CCK-8）assay.The expressions of Bcl-2,topoisomeraseⅡ（topoⅡ）,GDNF and LRIG1 were detected by western blotting.ResultsThe plasmids with LRIG1,LRIG1 siRNA,Bcl-2 or GDNF were successfully constructed and transfected into U251cells.The cell activities significantly decreased in LRIG1-transfected U251 cells compared with those without（P〈0.05）.The expressions of Bcl-2,topoⅡand GDNF significantly decreased in LRIG1-transfected U251 cells compared with those without（P〈0.05）,but the expressions of Bcl-2,topoⅡand GDNF significantly increased in LRIG1 siRNA-transfected U251 cells compared with those without（P〈0.05）.The expression of LRIG1 significantly decreased in Bcl-2-or GDNF-transfected U251 cells compared with those without（P〈0.05）.ConclusionsThe LRIGl can significantly inhibit cell proliferation of glioma cell line U251 cells.It may be by negatively regulating expressions of Bcl-2,topoⅡand GDNF.
出处
《中国临床神经外科杂志》
2013年第10期606-609,共4页
Chinese Journal of Clinical Neurosurgery
基金
国家自然基金资助项目(30973072)
