AUTHOR=Li Changjian , Zou Xiaoming , Cai Qingxin , Li Jiacheng , Yang Shifeng , Zhang Ange , Chen Chongyan , Zhu Lei 

TITLE=Comprehensive Expression Profile Analysis of Neutrophil Extracellular Trap-Affected Genes in Gastric Cancer Cells and the Clinical Significance of lncRNA NEAT1-Related Signaling

JOURNAL=Frontiers in Oncology

VOLUME=12

YEAR=2022

URL=https://www.frontiersin.org/journals/oncology/articles/10.3389/fonc.2022.798531

DOI=10.3389/fonc.2022.798531

ISSN=2234-943X

ABSTRACT=<sec><title>Background</title><p>Gastric cancer (GC) is the fifth most common malignant tumor and the third leading cause of cancer-related deaths worldwide. Neutrophil extracellular traps (NETs) can enhance the invasion of GC cells and are associated with poor prognosis in patients. However, its mechanism of action is not completely understood.</p></sec><sec><title>Methods</title><p>The content of NETs in the peripheral blood of patients with GC was detected by enzyme-linked immunosorbent assay. GC AGS cells were treated with or without NETs for 24 h. High-throughput RNA sequencing was performed to screen differentially expressed long non-coding RNAs (lncRNAs), microRNAs (miRNAs), and messenger RNAs (mRNAs). Real-time polymerase chain reaction (PCR) was used to verify gene expression. A competing endogenous RNA (ceRNA) regulatory network was constructed. Modules were screened using the molecular complex detection (MCODE) plug-in. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed using the genes in the network. The role and clinical significance of the lncRNA <italic>NEAT1</italic>-related signaling pathway were validated.</p></sec><sec><title>Results</title><p>The content of NETs in the patients with GC was significantly higher than that in healthy controls and was also higher in patients with high-grade (stages III and IV) GC. NETs promoted the invasion of AGS cells. A total of 1,340 lncRNAs, 315 miRNAs, and 1,083 mRNAs were differentially expressed after NET treatment. The expression of five genes was validated using real-time PCR, which were in accordance with the RNA sequencing results. A ceRNA regulatory network was constructed with 1,239 lncRNAs, 310 miRNAs, and 1,009 mRNAs. Four genes (<italic>RAB3B</italic>, <italic>EPB41L4B</italic>, <italic>ABCB11</italic>, and <italic>CCDC88A</italic>) in the ceRNA network were associated with patient prognosis, with <italic>RAB3B</italic> being the most prominent and with signaling among the lncRNA <italic>NEAT1</italic>, the miRNA miR-3158-5p, and <italic>RAB3B</italic>. <italic>NEAT1</italic> was upregulated in AGS cells after NET treatment. RNA interference of <italic>NEAT1</italic> inhibited the invasion of AGS cells induced by NETs, inhibited miR-3158-5p expression, and promoted <italic>RAB3B</italic> expression. <italic>NEAT1</italic> and <italic>RAB3B</italic> expression were positively correlated in patients with GC. Furthermore, <italic>RAB3B</italic> was upregulated and miR-3158-5p was downregulated in GC tissues compared with adjacent normal tissues, which was also associated with cancer stage.</p></sec><sec><title>Conclusion</title><p>This study provides a comprehensive analysis of differentially expressed genes in NET-treated GC cells and validated the clinical significance of <italic>NEAT1</italic>-related signaling.</p></sec>