LRIG1在胶质瘤细胞系U251细胞中的作用  

Effect of leucine-rich repeats and immunoglobulin-like domains 1 on glioma cell line U251

作　　者：刘宝辉[1] 陈谦学[1] 李明昌[1] 郭振涛[1] 朱晓楠[1] 冀保卫[1] 董慧敏[1] 吴立权[1] 田道锋[1] 易伟[1] 

机构地区：[1]武汉大学人民医院神经外科,武汉430060

出　　处：《中华神经医学杂志》2013年第10期980-985,共6页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金（30973072、81372683）;中国抗癌协会神经肿瘤专业委员会神经肿瘤研究项目~（CSNO-2013-MSD004）

摘　　要：目的探讨多亮氨酸重复区免疫球蛋白样蛋白I（LRIG1）在胶质瘤细胞系U251细胞中的作用及引起的相关基因变化。方法以PEGFP-LRIG1质粒转染体外常规培养的U251细胞，同时设PEGFP-N1组和空白组作为对照，RT-PCR检测3组细胞LRIG1、GDNF、拓扑异构酶Ⅱ（TOPOII）基因表达的变化。Westernblotting检测细胞LRIGl蛋白的表达；绘制PEGFP-N1和PEGFP-LRIGl组细胞1-5d的生长曲线并计算替莫唑胺对2组细胞的抑制率。应用siGDNF和siLRIGl敲低U251细胞中GDNF和LRIG1的表达．同时设单纯siGDNF、siLRIGl组和空白组作为对照，比较细胞增殖率的变化。结果3组细胞LRIG1基因和蛋白、TOPOII和GDNF基因表达不同，差异有统计学意义（P〈0．05）；与PEGFP-N1组和空白组比较，PEGFP-LRIG1组细胞LRIG1基因和蛋白的表达较高，TOPOll和GDNF基因表达较低，差异均有统计学意义（P〈0．05）。细胞生长1～5d时，PEGFP—LRIGl组细胞计数低于PEGFP-N1组，差异有统计学意义俨〈0．05）。加入替莫唑胺培养后，PEGFP—LRIGl组细胞抑制率高于PEGFP-N1组，差异有统计学意义（P〈0．05）。Westernblotting结果显示应用siGDNF和siLRIG1敲低LRIGJ和GDNF成功。siLRIGl组细胞抑制率高于对照组，siLRIGl＋siGDNF组胞抑制率低于siLRIGl组，差异均有统计学意义（P〈0．05）。结论LRIG1可以通过调节GDNF的表达来调节U251细胞的生长：LRIG1可以抑制GDNF、TOPOU的表达，提高胶质瘤细胞对替莫唑胺的化疗敏感性。Objective To investigate the role of leucine-rich repeats and immunoglobulin-like domains 1 （LRIG1） in U251 cells and its gene changes. Methods The stable cell line over-expressed LRIG1 （PEGFP-LRIG1） was established; PEGFP-N1-U251 cells and blank-U251 cells were used as controls. The expression changes of LRIG1, GDNF and topoisomerase II （TOPOII） were detected by real time-PCR; Western blotting was employed to detect the LRIG1 protein expression; the growth curve of PEGFP-LRIG1-U251 cells and PEGFP-N1-U251 cells was drew from the 1n to 5th d, and the cell inhibition rate oftemozolomide in these two groups was calculated. And then, siGDNF＋siLRIG1 was performed to knock down the expressions of GDNF and LR1G1; besides that, siGDNF, siLRIG1 and/or blank control group were employed as controls to compare the proliferation index. Results The LRIG1 gene and protein expressions, GDNF and TOPOH gene expressions were significantly different among the three groups （P〈0.05）; the gene and protein expressions of LRIG1 were up-regulated and GDNF and TOPOI1 gene expressions were down-regulated in the stable cell line LRIG1-U251 as compared with those in the PEGFP-N1-U251 cells and blank-U521 cells （P〈0.05）. Cell counting in the stable cell line LRIG1-U251 was significantly lower than that in the PEGFP-N1-U251 cells on the 1-5 dof growth （P〈0.05）. After temozolomide was added, cell inhabitation rate in the PEGFP-LRIG1-U251 group was obviously higher that that in the PEGFP-N1-U251 group （P〈0.05）. SiGDNF＋siLRIG1 could successfully knock down the expressions of LRIG1 and GDNF; the inhabitation rate of siLRIG1 was significantly higher than that of blank control group, and the that of siGDNF＋siLRIG1 group was significantly lower than that in the siLRIG1 group （P〈0.05）. Conclusion LRIG1 can regulate proliferation of U251 cells by regulating GDNF. LRIG 1 can inhibit GDNF and TOPOII expressions and improve the chemosensitivity of U251 cells.

关 键 词：多亮氨酸重复区免疫球蛋白样蛋白1 胶质细胞源性神经生长因子 拓扑异构 酶Ⅱ U251细胞 细胞增殖 

分 类 号：R739.41[医药卫生—肿瘤]