摘要
目的:探讨肿瘤微环境中神经元对胶质瘤细胞迁移能力的影响。方法:分离培养胚胎18 d(E18)大鼠神经元,利用表达靶向神经连接蛋白3(NLGN3)分子shRNA的重组慢病毒(Lv-NLGN3-shRNA)感染神经元,real time RT-PCR和酶联免疫吸附试验(ELISA)检测NLGN3的表达,收集神经元培养上清与胶质瘤细胞系U251细胞共培养,Transwell实验检测U251细胞迁移能力,Western Blot方法检测U251细胞中哺乳动物雷帕霉素靶蛋白(mTOR)和基质金属蛋白酶-9(MMP-9)的表达。结果:Lv-NLGN3-shRNA重组慢病毒感染能够降低大鼠原代神经元中NLGN3 mRNA和蛋白的的表达,神经元培养上清与U251细胞共培养可以增加后者的迁移能力并上调mTOR和MMP-9的表达。然而,NLGN3表达被Lv-NLGN3-shRNA抑制后,培养上清对U251细胞促迁移能力下降,同时mTOR和MMP-9表达降低。结论:大鼠原代神经元可以通过NLGN3/mTOR/MMP-9信号通路增强U251细胞的迁移能力,这一途径可能是胶质瘤细胞转移的机制之一。
Objective:To investigate the effect of neurons on the migration ability of glioma cells in tumor microenvironment.Methods:Embryonic 18 d(E18)rat neurons were isolated and cultured,and infected with Lv-NLGN3-shRNA.The expression of NLGN3 was detected by real time RT-PCR and enzymed-linked immunosorenda assay(ELISA).Neuron culture supernatant was collected and co-cultured with glioma cell line U251 cells.The migration ability of U251 cells was detected by Transwell assay.The expressions of mammalian target of rapamycin(mTOR)and matrix metalloproteinase-9(MMP-9)in U251 cells were detected by Western Blot.Results:Infection with Lv-NLGN3-shRNA can reduce NLGN3 mRNA and protein expression in primary rat neurons,and co-culture of neuron culture supernatant with U251 cells can increase the migration ability and up-regulate the expression of mTOR and MMP-9.When the NLGN3 expression was inhibited by Lv-NLGN3-shRNA,the migration ability of U251 cells was decreased,and the expressions of mTOR and MMP-9 were down-regulated.Conclusion:The NLGN3/mTOR/MMP-9 signaling pathway of primary rat neurons can enhance the migration ability of U251 cells,which may be one of the mechanisms of glioma cell metastasis.
作者
雷伟
王诗邈
谢奥坦
朴浩哲
LEI Wei;WANG Shimiao;XIE Aotan;PIAO Haozhe(Graduate School of Dalian Medical University,Dalian 116051;Department of Neurosurgery,Institute of Neuroscience,General Hospital of Northern Theater Command Base,Shenyang 110016;Liaoning cancer hospital&Institute,Shenyang 110801,China)
出处
《神经解剖学杂志》
CAS
CSCD
2023年第1期97-102,共6页
Chinese Journal of Neuroanatomy
基金
辽宁省“兴辽英才计划”(XLYC1902023)。