脑膜瘤组织中脆性X相关基因1过表达促进脑膜瘤细胞生长与血管生成的研究  

Study on the Overexpression of Fragile X-Related Gene 1 in Meningioma Tissue to Promote the Growth and Angiogenesis of Meningioma Cells

作　　者：曹杰[1] 张杰[1] 孟发财[1] 蔺鹏帧 袁云超 黄卫东[1] 侯明山[1] 王继军[1] 缪星宇[1] 师蔚[2] 李民[1] CAO Jie;ZHANG Jie;MENG Facai(The People's Hospital of Shaanxi Province, Shaanxi Xi'an 710004, China)

机构地区：[1]陕西省人民医院神经外科,陕西西安710004 [2]西安交通大学第二附属医院神经外科,陕西西安710004

出　　处：《河北医学》2021年第6期910-916,共7页Hebei Medicine

基　　金：陕西省科技计划项目,(编号:2019SF-073)。

摘　　要：目的:探讨人脑膜瘤组织中RNA结合蛋白脆性X相关基因1(FXR1)表达情况及其对脑膜瘤细胞生长和血管生成的影响。方法:收集我院24例脑膜瘤患者的肿瘤组织及瘤旁组织标本,免疫组织化学染色检测FXR1表达。分离并培养人脑膜瘤细胞,将培养的细胞分为空白对照组、pcDNA3.1-NC组和pcDNA3.1-FXR1组,采用脂质体转染法将pcDNA3.1-FXR1质粒载体与pcDNA3.1空质粒载体转染至脑膜瘤细胞;实时荧光定量PCR和蛋白质印迹(Western blot)检测细胞中FXR1 mRNA和蛋白表达水平;CCK-8实验检测细胞增殖活性;细胞克隆形成实验检测细胞菌落形成能力;Western blot检测VEGF、EDN-1蛋白表达水平;流式细胞术检测血管内皮细胞的标记CD31与CD34的阳性细胞表达;体外小管生成试验检测HUVECs成管情况。结果:脑膜瘤组织中FXR1阳性表达较瘤旁组织显著增加(P<0.05)。与空白对照组和pcDNA3.1-NC组比较,pcDNA3.1-FXR1组细胞中FXR1 mRNA和蛋白相对表达量均显著升高(P<0.05),转染48、72和96 h后细胞增殖活性显著升高(P<0.05),细胞克隆形成数目显著增加(P<0.05),VEGF、EDN-1蛋白表达水平显著增加(P<0.05),CD34和CD31标记的阳性细胞数显著增多(P<0.05)同时,管腔形成数目明显增多。结论:FXR1在人脑膜瘤组织中呈高表达,过表达FXR1可促进脑膜瘤细胞生长与血管生成。Objective:To investigate the expression of RNA-binding gene fragile X-related protein 1(FXR1)in human meningiomas and its effect on the growth and angiogenesis of meningiomas.Methods:Collect the tumor tissue and adjacent tissue specimens of 24 meningiomas in our hospital,and detect the expression of FXR1 by immunohistochemical staining.Isolate and culture human meningioma cells,and divide the cultured cells into blank control group,pcDNA3.1-NC group and pcDNA3.1-FXR1 group.Transfect the pcDNA3.1-FXR1 plasmid vector and the pcDNA3.1 empty plasmid vector to meningiomas by lipofection;Real-time fluorescent quantitative PCR and Western blot detection of FXR1 mRNA and protein expression levels in cells were pweformed;CCK-8 experiment was conducted to detect cell proliferation activity;cell clone formation test was conducted to detect cell colony forming ability;Western blot was conducted to detect VEGF and EDN-1 protein expression levels;flow cytometry was conducted to detect the positive cell expression of the markers CD31 and CD34 of vascular endothelial cells;in vitro tubule formation test was conducted to detect the tube formation of HUVECs.Results:The positive expression of FXR1 in meningioma tissue was significantly higher than that in adjacent tissues(P<0.05).Compared with the blank control group and the pcDNA3.1-NC group,the relative expression of FXR1 mRNA and protein in the cells of the pcDNA3.1-FXR1 group were significantly increased(P<0.05),cell proliferation activity increased significantly after 48,72 and 96 hours of transfection(P<0.05),the number of cell clone formation increased significantly(P<0.05),VEGF and EDN-1 protein expression levels increased significantly(P<0.05),the number of positive cells marked by CD34 and CD31 increased significantly(P<0.05),at the same time,the number of lumens formed increased significantly.Conclusion:FXR1 is highly expressed in human meningiomas.Overexpression of FXR1 can promote meningioma cell growth and angiogenesis.

关 键 词：脑膜瘤 脆性X相关基因1 增殖 血管生成 

分 类 号：R73[医药卫生—肿瘤]