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核仁纺锤体相关蛋白 1(NUSAP1)通过调节细胞周期蛋白依赖性激酶(CDK1)和 DLGAP5 的表达抑制浸润性乳腺癌细胞增殖并增强对表柔比星的敏感性。

Nucleolar and Spindle Associated Protein 1 (NUSAP1) Inhibits Cell Proliferation and Enhances Susceptibility to Epirubicin In Invasive Breast Cancer Cells by Regulating Cyclin D Kinase (CDK1) and DLGAP5 Expression.

机构信息

Department of Oncology, The Third Xiangya Hospital of Central South University, Changsha, Hunan, China (mainland).

出版信息

Med Sci Monit. 2018 Nov 26;24:8553-8564. doi: 10.12659/MSM.910364.

Abstract

BACKGROUND Differentially expressed genes (DEGs) of IBC were selected from the Gene Expression Omnibus (GEO) chip data: GSE21422 and GSE21974. Network analysis of the DEGs and IBC-related genes was performed in STRING database to find the core gene. Thus, this study aimed to determine the role of NUSAP1 in invasive breast cancer (IBC) and to investigate its effect on drug susceptibility to epirubicin (E-ADM). MATERIAL AND METHODS The mRNA expression of NUSAP1 was determined by quantitative polymerase chain reaction (q-PCR). The protein expression was detected by Western blotting. Cell growth and growth cycle were detected by MTT assay and flow cytometry, respectively. Cell migration and invasion were tested by Transwell assay. RESULTS Through use of gene network analysis, we found that NUSAP1 interacts with IBC-related genes. NUSAP1 presented high expression in IBC tissue samples and MCF-7 cells. NUSAP1 overexpression promoted the growth, migration, and invasion of MCF-7 cells. While NUSAP1 gene silencing downregulated the expression of genes associated with cell cycle progression in G2/M phase, cyclin D kinase (CDK1) and DLGAP5 arrested cells in G2/M phase and significantly inhibited the growth, migration, and invasion of MCF-7 cells. si-NUSAP1 increased the susceptibility of MCF-7 cells to E-ADM-induced apoptosis. CONCLUSIONS Our study provides evidence that downregulation of NUSAP1 can inhibit the proliferation, migration, and invasion of IBC cells by regulating CDK1 and DLGAP5 expression and enhances the drug susceptibility to E-ADM.

摘要

背景

从基因表达综合数据库(GEO)芯片数据 GSE21422 和 GSE21974 中选择 IBC 的差异表达基因(DEGs)。在 STRING 数据库中对 DEGs 和与 IBC 相关的基因进行网络分析,以找到核心基因。因此,本研究旨在确定 NUSAP1 在浸润性乳腺癌(IBC)中的作用,并研究其对表阿霉素(E-ADM)敏感性的影响。

材料和方法

通过定量聚合酶链反应(q-PCR)测定 NUSAP1 的 mRNA 表达。通过 Western blot 检测蛋白表达。通过 MTT 测定和流式细胞术分别检测细胞生长和生长周期。通过 Transwell 测定检测细胞迁移和侵袭。

结果

通过使用基因网络分析,我们发现 NUSAP1 与 IBC 相关基因相互作用。NUSAP1 在 IBC 组织样本和 MCF-7 细胞中呈现高表达。NUSAP1 过表达促进 MCF-7 细胞的生长、迁移和侵袭。而 NUSAP1 基因沉默下调 G2/M 期细胞周期进展相关基因的表达,细胞周期蛋白依赖性激酶 1(CDK1)和 DLGAP5 使细胞停滞在 G2/M 期,并显著抑制 MCF-7 细胞的生长、迁移和侵袭。si-NUSAP1 增加了 MCF-7 细胞对 E-ADM 诱导凋亡的敏感性。

结论

我们的研究提供了证据,表明下调 NUSAP1 通过调节 CDK1 和 DLGAP5 的表达可以抑制 IBC 细胞的增殖、迁移和侵袭,并增强对 E-ADM 的药物敏感性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/334e/6278864/34bc91bac2f2/medscimonit-24-8553-g001.jpg

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