NRG4基因对胶质瘤细胞的增殖、迁移及侵袭的影响  

Effect of NRG4 on proliferation,migration,and invasion of glioma cells

作　　者：舒波 出良钊 甘鸿川 黎浪 张胜 赵进城 SHU Bo;CHU Liangzhao;GAN Hongchuan;LI Lang;ZHANG Sheng;ZHAO Jincheng(Department of Neurosurgery,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Neurosurgery,the Second People's Hospital of Guiyang,Guiyang 550081,Guizhou,China)

机构地区：[1]贵州医科大学附属医院神经外科,贵州贵阳550004 [2]贵阳市第二人民医院神经外科,贵州贵阳550081

出　　处：《贵州医科大学学报》2024年第4期508-514,共7页Journal of Guizhou Medical University

基　　金：国家自然科学基金(81560409)。

摘　　要：目的探讨神经调节蛋白4(NRG4)对神经胶质瘤细胞的增殖、迁移以及侵袭作用的影响及机制。方法采用逆转录-定量聚合酶链反应(RT-qPCR)法检测神经胶质瘤患者(研究组)、对照组(同期健康志愿者)血清以及星形胶质细胞HA、神经胶质瘤细胞系U251、SHG44、LN229及T98G中NRG4和人表皮生长因子受体4(ErbB4)mRNA表达水平;将U251细胞分为Control组(空白对照)、NC组(转染不带RNA的质粒)、si-NRG4组(转染si-NRG4)和si-NRG4+ErbB4组[转染si-NRG4和ErbB4过表达质粒(pcDNA-ErbB4)]4组,通过细胞计数试剂盒-8(CCK-8)检测细胞增殖情况,划痕愈合实验和Transwell实验分析U251细胞的迁移与侵袭能力变化,免疫印迹(Western blot)检测PI3K/AKT通路中关键蛋白磷脂酰肌醇-3-激酶(PI3K)中p110α、p110β亚型、磷酸化丝氨酸/苏氨酸蛋白激酶(pAKT)中ser473和thr308 a以及蛋白激酶(AKT)表达水平。结果研究组血清中NRG4和ErbB4 mRNA表达量明显高于对照组(P<0.0001),神经胶质瘤细胞系U251中NRG4和ErbB4 mRNA相对表达量最高;与NC组比较,si-NRG4组中NRG4基因表达下调,不仅抑制U251细胞的增殖、迁移与侵袭,还降低了PI3 K-p110α,pAKT-ser473 and pAKT-thr308蛋白表达(P<0.05)。结论抑制NRG4表达可以减缓磷脂酰肌醇有关的信号通路PI3K/AKT通路的激活,抑制U251细胞的增殖和迁移,促进U251细胞凋亡,因此NRG4有可能成为治疗胶质瘤的潜在靶点。Objective To explore the effect of neuregulin 4(NRG4)on proliferation,migration and invasion of glioma cells.Methods Reverse transcription-quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression levels of NRG4 and human epidermal growth factor receptor 4(ErbB4)in the sera of glioma patients(study group)and healthy volunteers at the period(control group)as well as in glioma cell lines U251,SHG44,LN229,and T98G.U251 cells were divided into Control group(blank control),NC group(transfection with plasmid without carrying RNA),si-NRG4 group(transfection with si-NRG4),and si-NRG4+ErbB4 group[transfection with si-NRG4+overexpressing plasmids(pcDNA-ErbB4)].Cell counting kit-8(CCK-8)kit was used to assay cell proliferation.Wound healing and Transwell experiments were used to analyze the changes in U251 cell migration and invasion capacity.Western blot was used to detect the expression levels of key proteins such as phosphatidylinositol-3(PI3K)subunits p110α,p110β,phospho protein kinase B(pAKT;ser473 and thr308a),and AKT in PI3K/AKT pathway.Results The mRNA expression levels of NRG4 and ErbB4 in study group were significantly higher than those in control group(P<0.0001).The relative expression levels of NRG4 and ErbB4 mRNA were the highest in glioma cell line U251 among the above glioma cell lines.When compared with NC group,NRG 4 gene expression was downregulated in si-NRG 4 group,leading to not only inhibiting cell proliferation,migration and invasion of U251 cells,but also reducing the protein expression levels of PI3K-p110α,pAKT-ser473,and pAKT-thr308(P<0.05).Conclusion Inhibiting NRG4 expression may suppress the activation of PI3K/AKT pathway,U251 cell proliferation and migration,promote U251 cellular apoptosis.Therefore,NRG4 may be a potential target for glioma therapy.

关 键 词：NRG4 ERBB4 PI3K/AKT 神经胶质瘤细胞 增殖 迁移 侵袭 

分 类 号：R739.41[医药卫生—肿瘤]