神经降压素及其受体1对胶质瘤细胞侵袭的影响  

Effect of neurotensin and neurotensin receptor 1 on migration and invasion of human glioma cells

作　　者：欧阳庆[1] 周济[1] 易良[1] 徐伦山[1] 许民辉[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所神经外科,重庆400042

出　　处：《第三军医大学学报》2014年第24期2427-2431,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金青年-面上连续资助项目(81270039);中国博士后科学基金面上一等资助项目(2013M530388);重庆市博士后科研项目特别资助项目(Xm201341)~~

摘　　要：目的探讨神经降压素(neurotensin,NTS)及其受体1(neurotensin receptor 1,NTSR1)对恶性胶质瘤细胞迁移和侵袭能力的影响。方法体外实验分为空白对照组,NTS刺激组,NTS+sc-shRNA组和NTS+sh NTSR1组,采用慢病毒介导的RNA干扰技术沉默人胶质瘤细胞系U87中NTSR1的表达后采用细胞划痕实验和Transwell侵袭实验检测NTS/NTSR1对胶质瘤细胞迁移和侵袭能力的影响;Western blot检测NTS/NTSR1对N-钙粘素(N-cadhein)和基质金属蛋白酶-9(MMP-9)蛋白表达水平的影响。在体实验分为实验组sh NTSR1-U87和对照组sc-shRNA-U87,建立裸鼠颅内移植瘤模型。利用小动物磁共振技术(MRI)监测裸鼠颅内胶质瘤的生长,评估瘤细胞对瘤周组织的侵袭情况以及NTS/NTSR1对荷瘤鼠生存期的影响。结果 NTS/NTSR1促进恶性胶质瘤细胞株U87的体外迁移和侵袭能力,划痕结果显示,NTS刺激组细胞迁移率[(38.02±6.87)%高于空白对照组(13.46±4.49)%,P<0.05]。NTS+sh NTSR1组细胞迁移率(9.48±3.08)%低于NTS+sc-shRNA组[(35.48±12.89)%,P<0.05]。Transwell结果也证实,NTS刺激组穿膜细胞光密度值(0.25±0.02)高于空白对照组[(0.16±0.02),P<0.05]。NTS+sh NTSR1组光密度值(0.15±0.03)低于NTS+sc-shRNA组光密度值[(0.24±0.04),P<0.05]。NTS/NTSR1上调U87细胞MMP-9的表达并抑制N-钙粘素的表达(P<0.05)。在体实验结果显示,沉默NTSR1抑制荷瘤小鼠颅内胶质瘤的生长和侵袭,延长荷瘤鼠生存期(P<0.05)。结论抑制NTSR1的表达能降低瘤细胞体内外侵袭能力,延长小鼠的生存期。Objective To investigate the effect of neurotensin（NTS） and neurotensin receptor 1（NTSR1） on the migration and invasion of human glioma U87 cells.Methods An NTSR1-targeting shRNA vector and a non-homologous scramble sequence were constructed to transfect human glioma U87 cells,and the positive clones were screened with puromycin and then named as sh NTSR1 and sc-shRNA.The expression levels of NTSR1,N-cadherin and MMP-9 were detected by Western blotting.The cell migration and invasion ability was determined by wound scratch assay and Transwell invasion assay.After the intracranial transplantation tumor model was established,magnetic resonance imaging（MRI） was used to measure the tumor volume,and the protein levels of MMP-9 and N-cadherin in the orthotopic tumor tissue were evaluated by Western blotting.Results The protein level of NTSR1 was significantly decreased after NTSR1 knockdown,and the cell migration and invasion ability was markedly reduced（P 〈 0.05）.The wound scratch assay results showed that the cell migration rate（38.02 ± 6.87） % of the NTS stimulation group was higher than that（13.46 ±4.49） % of the blank control group（P 〈 0.05）,and the cell migration rate（9.48 ± 3.08） of the NTS ＋sh NTSR1 group was lower than that（35.48 ± 12.89） of the NTS ＋ sc-shRNA group（P 〈 0.05）.The Transwell invasion assay results also showed that the OD value（0.25 ± 0.02） of the NTS stimulation group was higher than that（0.16 ± 0.02） of the blank control group（P 〈 0.05）,and the OD value（0.15 ± 0.03） of the NTS ＋ sh NTSR1 group was lower than that（0.24 ± 0.04） of the NTS ＋ sc-shRNA group（P 〈 0.05）.The Western blotting results showed that MMP-9 expression was increased,however,the level of N-cadherin was decreased by NTS stimulation.Conversely,the expression of N-cadherin was increased,however,the level of MMP-9 was decreased after NTSR1 knockdown.More importantly,the in vivo results showed that NTSR1 downregulation could impair U87 cell inv

关 键 词：胶质瘤 神经降压素 神经降压素受体1 侵袭 

分 类 号：R341.6[医药卫生—基础医学] R73-37