EGFR信号通路调控人胶质瘤U87细胞侵袭转移的分子机制  

Molecular mechanism of EGFR signaling pathway mediating invasion and metastasis of U87 glioma cell line

作　　者：邢细红[1,2] 曾晖[1] 王雄伟[1,3] 郭东生[4] 汪雷[1] 万志先[1] 李焘[1] 周红建[1] 

机构地区：[1]三峡大学第一临床医学院神经外科,湖北宜昌443003 [2]荆州市第二人民医院 [3]宜昌市中心人民医院神经外科 [4]华中科技大学同济医学院附属同济医院,武汉430030

出　　处：《重庆医学》2012年第18期1818-1820,I0001,共4页Chongqing medicine

摘　　要：目的探讨表皮生长因子受体(EGFR)信号通路与人胶质瘤细胞(U87细胞)增殖和侵袭转移的关系及调控分子机制。方法表皮生长因子(EGF,100ng/mL)、EGFR抑制剂———AG1478(10μmol/L)单独和联合处理U87细胞,采用MTT法和Transwell小室体外侵袭实验检测U87细胞增殖和体外侵袭能力;明胶酶谱法检测基质金属蛋白酶-2(MMP-2)、MMP-9的表达水平;Western blot法检测磷酸化EGFR(P-EGFR)、磷酸化AKT(P-AKT)蛋白表达。结果 EGF(100ng/mL)增加P-EGFR和P-AKT蛋白表达,使加药后24h和48h的细胞生长率分别提高了19.25%、22.32%(P<0.05),使12h过膜细胞数由(27±4)个上升到(126±3)个(P<0.05),促进U87细胞的MMP-2、MMP-9蛋白表达(P<0.05);AG1478(10μmol/L)可以阻断EGF增加P-EGFR和P-AKT蛋白表达的作用(P<0.05),并具有时间依赖性(P<0.05),减弱U87细胞体外侵袭能力(P<0.05),抑制MMP-2、MMP-9蛋白的表达(P<0.05)。结论 EGFR-PI3K/AKT信号通路参与调节U87细胞增殖和侵袭转移过程,其机制可能是EGFR-PI3K/AKT信号通路活化后,导致MMP-2和MMP-9蛋白的表达增加,对细胞外基质的破坏增强。Objective To study the relationship between EGFR signal pathway and invasion and metastasis of U87 glioma cells, and discuss the molecular mechanism. Methods U87 glioma cells were cultured in medium that contained epidermal growth factor （EGF 100 ng/mL） or epidermal growth factor inhibitor AG1478（10 μmol/L） or combination,then the methyl thiazolyl tetrazo- lium （MTT） assay and transwell chamber were used to detect the proliferation and invasive ability of U87 glioma cells;Expression levels of matrix metalloproteinases-2 （MMP-2）, matrix metalloproteinases-9 （MMP-9） were determined by gelatinase zymography e- lectrophoresis Western blot was used to determine the expression levels of phosphorylation of the epidermal growth factor receptor （P-EGFR） ,and phosphorylation of protein kinase B （P-AKT）. Results Exogenous EGF（100 ng/mL）increased the expression lev- els of P-EGFR.P-AKT,the growth ratio increased 19.25% ,22.32% （P〈0.05）at 24,48 h respectively after treated,increased the number of invasion cell from （27 4-4） ceils to （126± 3） ceils （P〈0.05）, and increased the expression levels of MMP-2, MMP-9 AG1478 could block the effects of EGF increased the expressions of P-EGFR, P-AKT in time-independent（P〈0.05）, decreased the invasive ability of U87 glioma（P^0.05） ,inhibited the expression of MMP-2,MMP-9（P〈0.05）. Conclusion The EGFR-PI3K/ AKT signaling pathways involved in the regulation of U87 glioma cells proliferation and invasion and metastasis. Its mechanism is possible that after the EGFR-PI3 K/AKT signaling pathways activated, caused the high expression of MMP-2, MMP-9 and increased the damage to the exracellular matrix（ECM）.

关 键 词：神经胶质瘤 受体 表皮生长因子 基质金属蛋白酶类 肿瘤转移 肿瘤细胞 培养的 

分 类 号：R739.4[医药卫生—肿瘤]