[1]丁　昊　周志伟　易　伟　雷　霆　简志宏　刘仁忠.LRIG1相关功能片段对EGFR活性和胶质瘤细胞增殖的影响[J].中国临床神经外科杂志,2015,(09):544-546，575.[doi:10.13798/j.issn.1009-153X.2015.09.011]
　DING Hao,ZHOU Zhi-wei,YI Wei,et al.Effects of functional regions of LRIG1 on signaling of EGFR and cell proliferation of gliomas[J].,2015,(09):544-546，575.[doi:10.13798/j.issn.1009-153X.2015.09.011]
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LRIG1相关功能片段对EGFR活性和胶质瘤细胞增殖的影响()
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《中国临床神经外科杂志》[ISSN:1009-153X/CN:42-1603/TN]

卷:
期数:
2015年09期
页码:
544-546，575
栏目:
论著
出版日期:
2015-09-30

文章信息/Info

Title:
Effects of functional regions of LRIG1 on signaling of EGFR and cell proliferation of gliomas
文章编号:
1009-153X（2015）09-0544-03
作者:
丁　昊　周志伟　易　伟　雷　霆　简志宏　刘仁忠
430060 武汉，武汉大学人民医院神经外科（丁　昊、周志伟、易　伟、简志宏、刘仁忠）；430030 武汉，华中科技大学同济医学院附属同济医院神经外科（雷　霆）
通讯作者：易　伟，E-mail：yiwtj@hotmail.com
Author(s):
DING Hao1;  ZHOU Zhi-wei1;  YI Wei1;  LEI Ting2;  JIAN Zhi-hong1;  LIU Ren-zhong1.
1. Department of Neurosurgery, Renmin Hospital, Wuhan University, Wuhan 430060, China; 2. Department of Neurosurgery, Tongji Hospital, Tongji Medical School, Huazhong University of Sciences and Technology, Wuhan 430030, China
关键词:
胶质瘤; 多亮氨酸重复区免疫球蛋白样蛋白1; 表皮细胞生长因子受体; 细胞增殖
Keywords:
Leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1);  Extracellular domains;  Cytoplasmic domains;  Epidermal growth factor receptor (EGFR);  Cell proliferation
分类号:
R 739.41; Q 789
DOI:
10.13798/j.issn.1009-153X.2015.09.011
文献标志码:
A
摘要:
目的 探讨人多亮氨酸重复区免疫球蛋白样蛋白1（LRIG1）相关功能片段对表皮生长因子受体（EGFR）下游信号通路的影响及对人脑胶质瘤细胞增殖活性的影响。方法 构建缺失LRIG1胞内段（LRIG1-ET）的和缺失LRIG1胞外段（LRIG1-TC）的真核表达质粒，将这两个质粒及LRIG1全长（LRIG1-FL）质粒分别转染脑胶质瘤U251细胞系和原代星形胶质细胞瘤细胞，用蛋白质印迹技术检测EGFR下游信号蛋白有丝分裂原活化蛋白激酶激酶（MAPKK/MEK）的磷酸化水平，用MTT法检测转染细胞的增殖活性。结果 全长LRIG1、LRIG1胞外段、LRIG1胞内段对人脑U251细胞和原代胶质瘤细胞的增殖活性均有抑制作用；全长LRIG1蛋白的抑制作用最强，其次为胞内段，再次为胞外段。全长LRIG1、LRIG1胞外段、LRIG1胞内段均使原代星形胶质细胞瘤细胞的MEK蛋白磷酸化水平下降。结论 LRIG1全长、胞外段、胞内段均能通过抑制EGFR下游信号抑制U251细胞和原代星形胶质瘤细胞的细胞增殖；LRIG1胞外段和胞内段具有独立的功能，均有可能对人脑胶质瘤的治疗发挥重要作用。
Abstract:
Objectives To investigate whether the sub-regions of leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1) interact with the downstream signaling of epidermal growth factor receptor (EGFR) and whether they inhibit cell proliferation of human gliomas. Methods Two chimeric eukaryotic expression vectors, pIRES2-LRIG1-Δcyto and pFLAG-LRIG1-Δecto were constructed. The former encoded LRIG1, but lacked the cytoplasmic region (LRIG1-ET). The latter encoded LRIG1, but lacked the extracellular region (LRIG1-TC). These two plasmids and the full length LRIG1 (LRIG1-FL) plasmid were transfected into human glioma U251 cells and primary astrocytoma cells. After the transfection, the phosphorylation levels of mitogen-activated protein kinas MAPK/EPK kinase MEK were detected by Western blot. The cell proliferation of the transfected cells was evaluated by MTT assay. Results Both the phosphorylated-MEK (p-MEK) and cell proliferation of the transfected cells were inhibited as compared with the non-transfected cells. For the primary astrocytoma cells, the p-MEK decreased by 13.0%, 34.3%, and 48.0% and the cell proliferation decreased by 26.0%, 32.5% and 39.0% respectively in the LRIG1-Δcyto, LRIG1-Δecto, and LRIG1-FL vectors transfected cells. The proliferation of U251 cells decreased by 30.4%, 39.1%, and 47.3% upon the three vectors transfection, respectively. Conclusions All the three forms of LRIG1 are capable of suppressing cell proliferation of gliomas via down-regulating signaling of EGFR. Both the extracellular domains and the cytoplasmic domains of LRIG1 have independent biological functions, which give us an image of using these fragments for human gliomas treatments.

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备注/Memo

备注/Memo:
基金项目：国家自然科学基金（30973073；81172402；30271332）
更新日期/Last Update: 2015-09-30