NRP-1在胶质瘤细胞系中表达水平及亚细胞定位的鉴定  

Expression and subcellular localization of NRP-1 in glioma cell lines

作　　者：单煜恒 孙佳[1] 王玉梁[2] 梁之孔 龚燕华[2] 钟士江[1] 

机构地区：[1]武警后勤学院附属医院脑科医院,天津300162 [2]武警后勤学院分子生物学教研室,天津300309

出　　处：《武警后勤学院学报（医学版）》2016年第2期90-92,96,F0003,共5页Journal of Logistics University of PAP（Medical Sciences）

基　　金：天津市自然科学基金(13JCBJC24100);武警部队后勤部基金项目(WJHQ2012-13)

摘　　要：【目的】检测不同胶质瘤细胞系中神经纤毛蛋白1(neuropilin-1,NRP-1)的表达水平及亚细胞定位。【方法】传代培养人U87、U251、H4及大鼠C6胶质瘤细胞系,以人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVEC)为阳性对照、正常大鼠脑组织为阴性对照,采用RT-PCR、Western-blotting及细胞免疫荧光技术三种检测手段从信使RNA及蛋白层面分析NRP-1在四种胶质瘤细胞系中的表达及定位。【结果】四种胶质瘤细胞系(U87、U251、H4及C6)中均有NRP-1的阳性表达,且表达水平均明显高于正常大鼠脑组织(P<0.05);恶性程度最高的U87的NRP-1表达水平明显高于恶性程度较低的U251及H4(P<0.05);而U251及H4之间NRP-1的表达水平无明显差异(P>0.05);细胞免疫荧光结果显示NRP-1在四种胶质瘤细胞系中均有表达,且亚细胞定位主要分布于胞膜及胞质中。【结论】本实验确定了NRP-1在U87、U251、H4及C6胶质瘤细胞系中表达水平及亚细胞定位,为今后进一步将NRP-1应用于胶质瘤鉴别诊断与分级诊断奠定了理论基础。【Objective】To detect the expression and subcellular localization of neuropilin-1（NRP-1） in different glioma cell lines.【Methods】Glioma cell lines of human U87, U251 and H4 as well as rat C6 were subcultured. Human umbilical vein endothelial cells（HUVEC）was chosen as positive control, and normal rat brain were selected as negative control. The expression levels and subcellular localization of NRP-1 in the 4 glioma cell lines were detected with RT-PCR, Western-blotting and immunofluorescence staining.【Results】The expressions of NRP-1 were positive in the 4 glioma cell lines（U87, U251, H4 and C6） and the expression levels were significantly higher than that of normal rat brain（P〈0.05）. The expression level of NRP-1 in the most malignant U87 was significantly higher than those in U251 and H4 with less malignance（P〈0.05）; there was no distinct difference in NRP-1 expression between U251 and H4（P〈0.05）. The results of immunofluorescence staining showed that NRP-1 was expressed in the 4 glioma cell lines and its subcellular localization was mainly in membrane and cytoplasma.【Conclusion】Our results revealed the expression levels and subcellular localization of NRP-1 in the 4 glioma cell lines（U87, U251, H4 and C6）. It will provide a theoretical basis for the further application of NRP-1 in the differential diagnosis and grading of gliomas in the future.

关 键 词：胶质瘤 神经纤毛蛋白-1 亚细胞定位 

分 类 号：R739[医药卫生—肿瘤]