Houjun Zhou, Peng Bai
Neurosurgery Department 2, The Affiliated Hospital of Kunming Medical University, Kunming, Yunnan, China.
Neurosurgery Department 2, The Affiliated Hospital of Kunming Medical University, Kunming, Yunnan, China. Email:
Cell J. 2023 Apr 1;25(4):264-272. doi: 10.22074/cellj.2022.557561.1074.
This study was conducted to clarify the expression characteristics of cell cycle exit and neuronal differentiation 1 (CEND1) in glioma and its effects on the proliferation, migration, invasion, and resistance to temozolomide (TMZ) of glioma cells.
In this experimental study, CEND1 expression in glioma tissues and its relationship with patients' survival were analyzed through bioinformatics. Quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry were performed to detect CEND1 expression in glioma tissues. The cell counting kit-8 (CCK-8) method was adopted to detect cell viability and the effects of different concentrations of TMZ on the inhibition rate of glioma cell proliferation, and the median inhibitory concentration of TMZ (IC value) was calculated. 5-Bromo- 2'-deoxyuridine (BrdU), wound healing and Transwell assays were performed to evaluate the impacts of CEND1 on glioma cell proliferation, migration, and invasion. Besides, the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, Gene Ontology (GO) analysis, and Gene Set Enrichment Analysis (GSEA) were applied to predict the pathways regulated by CEND1. Nuclear factor-kappa B p65 (NF-κB p65) and phospho-p65 (p-p65) expression were detected by Western blot.
CEND1 expression was reduced in glioma tissues and cells, and its low expression was significantly associated with the shorter survival of glioma patients. CEND1 knockdown promoted glioma cell growth, migration, and invasion, and increased the IC50 value of TMZ, whereas up-regulating CEND1 expression worked oppositely. Genes co-expressed with CEND1 were enriched in the NF-κB pathway, and knocking down CEND1 facilitated p-p65 expression, while CEND1 overexpression suppressed p-p65 expression.
CEND1 inhibits glioma cell proliferation, migration, invasion, and resistance to TMZ by inhibiting the NF- κB pathway.
本研究旨在阐明胶质瘤中细胞周期退出与神经元分化1（CEND1）的表达特征及其对胶质瘤细胞增殖、迁移、侵袭和对替莫唑胺（TMZ）耐药性的影响。
在本实验研究中，通过生物信息学分析胶质瘤组织中CEND1的表达及其与患者生存的关系。采用定量实时聚合酶链反应（qRT-PCR）和免疫组织化学检测胶质瘤组织中CEND1的表达。采用细胞计数试剂盒-8（CCK-8）法检测细胞活力以及不同浓度TMZ对胶质瘤细胞增殖抑制率的影响，并计算TMZ的半数抑制浓度（IC值）。进行5-溴-2'-脱氧尿苷（BrdU）、伤口愈合和Transwell实验，以评估CEND1对胶质瘤细胞增殖、迁移和侵袭的影响。此外，应用京都基因与基因组百科全书（KEGG）分析、基因本体论（GO）分析和基因集富集分析（GSEA）预测CEND1调控的通路。通过蛋白质免疫印迹法检测核因子-κB p65（NF-κB p65）和磷酸化p65（p-p65）的表达。
CEND1在胶质瘤组织和细胞中的表达降低，其低表达与胶质瘤患者较短的生存期显著相关。敲低CEND1可促进胶质瘤细胞生长、迁移和侵袭，并增加TMZ的IC50值，而上调CEND1表达则产生相反的作用。与CEND1共表达的基因在NF-κB通路中富集，敲低CEND1促进p-p65表达，而CEND1过表达则抑制p-p65表达。
CEND1通过抑制NF-κB通路抑制胶质瘤细胞增殖、迁移、侵袭及对TMZ的耐药性。
