Wu Chunli, Dong Bo, Huang Lan, Liu Yafei, Ye Guanchao, Li Shihao, Qi Yu
Department of Thoracic Surgery, The First Affiliated Hospital of Zhengzhou University, Henan, China.
Biotherapy Center, The First Affiliated Hospital of Zhengzhou University, Henan, China.
Front Oncol. 2021 Oct 20;11:754290. doi: 10.3389/fonc.2021.754290. eCollection 2021.
The roles played by β-III-spectrin, also known as spectrin beta, non-erythrocytic 2 (), in the occurrence and development of lung adenocarcinoma (LUAD) have not been previously examined. Our study aimed to reveal the relationship between the expression and LUAD.
Twenty pairs of LUAD tissues and adjacent tissues were collected from patients diagnosed and treated at the Thoracic Surgery Department of The First Affiliated Hospital of Zhengzhou University from July 2019 to September 2020. RNA sequencing (RNA-seq) analysis determined that the expression of was higher in LUAD samples than in adjacent normal tissues. The expression levels of were examined in various databases, including the Cancer Cell Line Encyclopedia (CCLE), Gene Expression Omnibus (GEO), and Human Protein Atlas (HPA). The Search Tool for the Retrieval of Interacting Genes (STRING) online website was used to examine protein-protein interactions involving , and the results were visualized by Cytoscape software. The Molecular Complex Detection (MCODE) plug-in for Cytoscape software was used to identify functional modules of the obtained protein-protein interaction (PPI) network. Gene enrichment analysis was performed, and survival analysis was conducted using the Kaplan-Meier plotter. The online prediction website TargetScan was used to predict SPTBN2-targeted miRNA sequences by searching for sequences. Finally, we verified the expression of in the obtained tissue samples using real-time fluorescence quantitative polymerase chain reaction (RT-qPCR). The human lung cancer cell lines A549 and H1299 were selected for the transfection of small interfering RNA (siRNA) targeting (si-SPTBN2), and the knockdown efficiency was evaluated by RT-qPCR. The cellular proliferation, migration, and invasion capacities of A549 and H1299 cells were determined using the cell counting kit-8 (CCK-8) proliferation assay; the wound-healing assay and the Transwell migration assay; and the Matrigel invasion assay, respectively.
The expression of in non-small cell lung cancer (NSCLC) ranked 13th among cancer cell lines based on the CCLE database. At the mRNA and protein levels, the expression levels of were higher in LUAD tissues than in normal lung tissues. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that proteins related to were enriched in apoptotic and phagosomal pathways. Kaplan-Meier survival analysis revealed that expression was significantly related to the prognosis of patients with LUAD. The TargetScan database verified that miR-16 was a negative regulator of mRNA expression. The results of the CCK-8 cell proliferation assay revealed that knockdown significantly inhibited the cell proliferation abilities of A549 and H1299 cells. The wound-healing assay indicated that knockdown resulted in reduced migration after 48 h compared with the control group. The Transwell migration and invasion test revealed that the migration and invasion abilities were greatly decreased by knockdown compared with control conditions.
We uncovered a novel gene, , that was significantly upregulated in LUAD tissues relative to normal tissue expression. is highly expressed in LUAD, positively correlated with poor prognosis, and can promote the proliferation, migration, and invasion of LUAD cells.
β-III-血影蛋白（也称为血影蛋白β，非红细胞型2，即SPTBN2）在肺腺癌（LUAD）发生发展中的作用此前尚未得到研究。本研究旨在揭示SPTBN2表达与LUAD之间的关系。
收集2019年7月至2020年9月在郑州大学第一附属医院胸外科确诊并接受治疗的患者的20对LUAD组织及癌旁组织。RNA测序（RNA-seq）分析确定，SPTBN2在LUAD样本中的表达高于癌旁正常组织。在多个数据库中检测SPTBN2的表达水平，包括癌症细胞系百科全书（CCLE）、基因表达综合数据库（GEO）和人类蛋白质图谱（HPA）。利用在线网站搜索相互作用基因的工具（STRING）检测涉及SPTBN2的蛋白质-蛋白质相互作用，并通过Cytoscape软件将结果可视化。使用Cytoscape软件的分子复合物检测（MCODE）插件识别所得蛋白质-蛋白质相互作用（PPI）网络的功能模块。进行基因富集分析，并使用Kaplan-Meier生存分析绘图仪进行生存分析。通过搜索SPTBN2序列，利用在线预测网站TargetScan预测靶向SPTBN2的miRNA序列。最后，我们使用实时荧光定量聚合酶链反应（RT-qPCR）验证所得组织样本中SPTBN2的表达。选择人肺癌细胞系A549和H1299进行靶向SPTBN2的小干扰RNA（siRNA，即si-SPTBN2）转染，并通过RT-qPCR评估敲低效率。分别使用细胞计数试剂盒-8（CCK-8）增殖试验、伤口愈合试验和Transwell迁移试验以及基质胶侵袭试验测定A549和H1299细胞的细胞增殖、迁移和侵袭能力。
基于CCLE数据库，SPTBN2在非小细胞肺癌（NSCLC）细胞系中的表达排名第13位。在mRNA和蛋白质水平上，SPTBN2在LUAD组织中的表达水平高于正常肺组织。京都基因与基因组百科全书（KEGG）通路分析显示，与SPTBN2相关的蛋白质在凋亡和吞噬体通路中富集。Kaplan-Meier生存分析显示，SPTBN2表达与LUAD患者的预后显著相关。TargetScan数据库验证miR-16是SPTBN2 mRNA表达的负调节因子。CCK-8细胞增殖试验结果显示，敲低SPTBN2显著抑制A549和H1299细胞的细胞增殖能力。伤口愈合试验表明，与对照组相比，敲低SPTBN2导致48小时后迁移减少。Transwell迁移和侵袭试验显示，与对照条件相比，敲低SPTBN2使迁移和侵袭能力大大降低。
我们发现了一个新基因SPTBN2，其在LUAD组织中的表达相对于正常组织表达显著上调。SPTBN2在LUAD中高表达，与不良预后呈正相关，并且可促进LUAD细胞的增殖、迁移和侵袭。
