Nucleolar and spindle-associated protein 1 (NUSAP1) is a potential molecular marker and intervention target for glioblastoma (GBM). In this study, we aim to investigate upstream regulatory lncRNAs and miRNAs of NUSAP1 through both experimental and bioinformatic methods. We screened upstream lncRNAs and miRNAs of NUSAP1 through multiple databases based on ceRNA theory. Then, in vitro and in vivo experiments were performed to elucidate the relevant biological significance and regulatory mechanism among them. Finally, the potential downstream mechanism was discussed. LINC01393 and miR-128-3p were screened as upstream regulatory molecules of NUSAP1 by TCGA and ENCORI databases. The negative correlations among them were confirmed in clinical specimens. Biochemical studies revealed that overexpression or knockdown of LINC01393 respectively enhanced or inhibited malignant phenotype of GBM cells. MiR-128-3p inhibitor reversed LINC01393 knockdown-mediated impacts on GBM cells. Then, dual-luciferase reporter assay and RNA immunoprecipitation assay were conducted to validate LINC01393/miR-128-3p/NUSAP1 interactions. In vivo, LINC01393-knockdown decreased tumor growth and improved mice survival, while restoration of NUSAP1 partially reversed these effects. Additionally, enrichment analysis and western blot revealed that the roles of LINC01393 and NUSAP1 in GBM progression were associated with NF-κB activation. Our findings showed that LINC01393 sponged miR-128-3p to upregulate NUSAP1, thereby promoting GBM development and progression via activating NF-κB pathway. This work deepens understanding of GBM mechanisms and provides potential novel therapeutic targets for GBM.

中文翻译：

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LINC01393，一种新型长链非编码 RNA，通过 MiR-128-3p/NUSAP1 轴促进胶质母细胞瘤细胞增殖、迁移和侵袭

核仁和纺锤体相关蛋白 1 (NUSAP1) 是胶质母细胞瘤 (GBM) 的潜在分子标记和干预靶点。在本研究中，我们旨在通过实验和生物信息学方法研究 NUSAP1 的上游调控 lncRNA 和 miRNA。我们基于ceRNA理论通过多个数据库筛选了NUSAP1的上游lncRNA和miRNA。然后，通过体外和体内实验阐明了它们之间的相关生物学意义和调控机制。最后，讨论了潜在的下游机制。LINC01393和miR-128-3p被TCGA和ENCORI数据库筛选为NUSAP1的上游调控分子。它们之间的负相关在临床标本中得到证实。生化研究表明，LINC01393 的过表达或敲低分别增强或抑制 GBM 细胞的恶性表型。MiR-128-3p 抑制剂逆转了 LINC01393 敲低介导的对 GBM 细胞的影响。然后，进行双荧光素酶报告基因测定和 RNA 免疫沉淀测定以验证 LINC01393/miR-128-3p/NUSAP1 相互作用。在体内，LINC01393 敲低降低了肿瘤生长并提高了小鼠存活率，而 NUSAP1 的恢复部分逆转了这些影响。此外，富集分析和蛋白质印迹显示 LINC01393 和 NUSAP1 在 GBM 进展中的作用与 NF-κB 激活有关。我们的研究结果表明，LINC01393 吸收 miR-128-3p 以上调 NUSAP1，从而通过激活 NF-κB 通路促进 GBM 的发展和进展。