1.Lanzhou University Second Hospital;2.School of life sciences, Northwestern Normal University
目的 探讨NOVA1对胶质母细胞瘤(Glioblastoma,GBM)增殖、迁移的调控作用。方法 我们使用生物信息学方法分析NOVA1敲减小鼠脑皮质和野生对照小鼠脑皮质的遗传数据,以筛选NOVA1基因缺失后的差异表达基因(DEGs)。从基因表达综合数据库(GEO)下载GSE69709数据集,并选择6个样本进行分析,共筛选出149个差异表达基因,其中筛选的差异表达基因Cdhr1、Pmfbp1分别参与细胞迁移能力、细胞骨架的改变的相关通路。通过构建NOVA1敲减慢病毒,转染GBM细胞系U87及U251 。WB验证转染效率后,进行CCK8、集落形成、划痕和transwell迁移试验,分析细胞增殖、迁移。基于脑立体定位技术,将胶质母细胞瘤C6显微注射入小鼠纹状体区,构建原位移植瘤模型,通过免疫荧光观察NOVA1在小鼠颅内分布及表达情况。结果 在小鼠颅内,NOVA1主要富集在神经纤维丰富的区域,中脑脑干等。NOVA1在胶质母细胞瘤组织中是广泛表达的。此外NOVA1敲减后显著抑制胶质瘤细胞的增殖、迁移。 结论：NOVA1基因编码的蛋白主要分布于小鼠中脑、脑干。NOVA1敲减后抑制胶质母细胞瘤的增殖、迁移能力,且有重塑部分GBM细胞骨架的能力。
the aim of this study was to investigate the regulation of proliferation and migration of glioblastoma (GBM) cells by nova1.SMethods We used a bioinformatics approach to analyze genetic data from the cerebral cortex of nova1 knockdown mice and the cerebral cortex of wild-type control mice to screen for differentially expressed genes (DEGs) following deletion of the nova1. The GSE69709 dataset was downloaded from the Gene Expression Omnibus (GEO) database, and six samples were selected for analysis, a total of 149 differentially expressed genes were selected, among which the screened differentially expressed genes cdhr1, pmfbp1 were involved in the related pathways of cell migration ability, cytoskeleton alteration, respectively. GBM cell lines U87 and U251 were transfected by construction of nova1 knockdown lentivirus. After verifying transfection efficiency, CCK8, colony formation, scratch and Transwell migration assays were performed to analyze cell proliferation and migration. Based on brain stereotaxic techniques, glioblastoma C6 was microinjected into the striatal region of mice to construct an orthotopic xenograft tumor model, and the intracranial distribution and expression of nova1 in mice were visualized by immunofluorescence. Results Within the mouse brain, nova1 is mainly enriched in areas rich in nerve fibers, midbrain brainstem, etc Nova1 is widely expressed in glioblastoma tissues. Furthermore, knockdown of nova1 significantly inhibited glioma cell proliferation and migration.SConclusion The protein encoded by the nova1 gene is mainly distributed in the midbrain and brainstem of mice. Knockdown of nova1 inhibits glioblastoma proliferation and migration and has the ability to remodel part of the GBM cytoskeleton.