| circ_0039411通过miR-423-5p/IL-8促进中性粒细胞胞外诱捕网的形成以促进胃癌细胞的侵袭 |
| 投稿时间：2024-11-23 修订日期：2025-02-23 点此下载全文 |
| 引用本文：徐琦,倪娇娇,李晶晶,韦青.circ_0039411通过miR-423-5p/IL-8促进中性粒细胞胞外诱捕网的形成以促进胃癌细胞的侵袭[J].医学研究杂志,2025,54(7):66-71, 90 |
| DOI：
10.11969/j.issn.1673-548X.2025.07.013 |
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| 基金项目:浙江省医药卫生科技计划项目(2022KY666) |
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| 中文摘要:目的 探讨circ_0039411通过miR-423-5p/白介素-8(interleukin-8,IL-8)调控中性粒细胞胞外诱捕网(neutrophil extracellular traps,NETs)对胃癌细胞的影响。方法 采用定量实时反转录聚合酶链反应(quantitative real-time reverse transcription polymerase chain reaction,qRT-PCR)检测胃癌细胞株(HGC-27、SNU-16和AGS)和人胃黏膜细胞(GSE-1)中circ_0039411和miR-423-5p的表达水平。选取HGC-27建立circ_0039411低表达模型,设立实验组(转染circ_0039411的siRNA)和空白对照组(转染对照的siRNA)。在实验组细胞中建立miR-423-5p低表达模型,设立inhibitor组(转染miR-423-5p的抑制物)和NC组(转染空载)。使用佛波肉豆蔻醋酸(phorbol 12-myristate 13-acetate,PMA)诱导NETs的形成,设立PMA组(经PMA诱导)和对照组(未经PMA诱导)。构建胃癌细胞与中性粒细胞的共培养模型。采用CCK-8检测胃癌细胞的活性,采用Transwell和划痕检测胃癌细胞的迁移和侵袭能力,采用酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测IL-8表达水平,采用免疫荧光检测NETs的形成。结果 胃癌细胞HGC-27、SNU-16和AGS中circ_0039411和IL-8的表达水平高于人胃黏膜细胞GSE-1,miR-423-5p的表达水平低于人胃黏膜细胞GSE-1,差异有统计学意义(P<0.05)。相比于空白对照组,实验组细胞的活性、迁移能力、侵袭能力及分泌的IL-8水平均较低,差异有统计学意义(P<0.05),NETs的形成减少。相比于NC组,inhibitor组细胞的活性、迁移能力、侵袭能力及分泌的IL-8水平均较高,差异有统计学意义(P<0.05),NETs的形成增加。相比于对照组,PMA组胃癌细胞的活性、迁移能力和侵袭能力均较高,差异有统计学意义(P<0.05)。结论circ_0039411能够通过miR-423-5p/IL-8轴促进NETs的形成,并进一步促进胃癌细胞的迁移和侵袭。 |
| 中文关键词:胃癌 circ_0039411 miR-423-5p IL-8 中性粒细胞胞外诱捕网 |
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| Circ_0039411 Regulates Neutrophil Extracellular Traps Through MiR-423-5p/IL-8 to Promote the Invasion of Gastric Cancer Cells. |
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| Abstract:Objective To explore the effect of circ_0039411 on gastric cancer cells by regulating neutrophil extracellular traps(NETs) through miR-423-5p/interleukin-8(IL-8). Methods We used quantitative real-time reverse transcription polymerase chain reaction(qRT-PCR) to detect the expression levels of circ_0039411 and miR-423-5p in gastric cancer cell lines(HGC-27, SNU-16 and AGS) and human gastric mucosa cell(GSE-1). We selected HGC-27 to establish a circ_0039411 low expression model, and set up Experimental group(transfected circ_0039411siRNA) and Blank control group(transfected control siRNA). We selected the cell of Experimental group to establish a miR-423-5p low expression model, and set up inhibitor group(transfected miR-423-5p inhibitor) and NC group(transfected empty). We used phorbol 12-myristate 13-acetate(PMA) to induce the formation of NETs in vitro, and established a PMA group(induced by PMA) and a control group(not induced by PMA). We used Transwell chamber to construct a co-culture model of gastric cancer cells and neutrophils. We used CCK-8 to detect the activity of gastric cancer cells, scratch and Transwell to detect the migration and invasion ability of gastric cancer cells, enzyme linked immunosorbent assay(ELISA) to detect the expression level of IL-8, and immunofluorescence to detect the formation of NETs. Results The expression levels of circ_0039411 and IL-8 in gastric cancer cells HGC-27, SNU-16 and AGS were higher than those of human gastric mucosa cells GSE-1, and the expression level of miR-423-5p was lower than that of GSE-1, and the differences were statistically significant(P<0.05). Compared with the Blank control group, the activity, migration and invasion ability of cells and the secretion of IL-8 in the Experimental group were decreased, and the differences were statistically significant(P<0.05), and the formation of NETs was reduced. Compared with NC group, the cell activity, migration and invasion ability and secreted IL-8 of inhibitor group were increased, and the differences were statistically significant(P<0.05), and the formation of NETs was promoted. Compared with the control group, the PMA group exhibited significantly higher activity, migration ability, and invasion ability in gastric cancer cells, with statistically significant differences(P<0.05). Conclusion circ_0039411 can promote the formation of NETs through the miR-423-5p/IL-8 axis, and further promote the migration and invasion ability of gastric cancer cells. |
| keywords:Gastric cancer Circ_0039411 MiR-423-5p IL-8 Neutrophil extracellular traps |
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