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罗哌卡因通过circSCAF11/miR-145-5p轴抑制胶质瘤细胞的增殖、迁移和侵袭,同时诱导氧化应激和细胞凋亡。

Ropivacaine Inhibits Cell Proliferation, Migration and Invasion, Whereas Induces Oxidative Stress and Cell Apoptosis by circSCAF11/miR-145-5p Axis in Glioma.

作者信息

Yin Danqin, Liu Li, Shi Zhengyuan, Zhang Lihui, Yang Yan

机构信息

Department of Anesthesiology, Danyang People's Hospital of Jiangsu, Danyang City, Jiangsu Province, People's Republic of China.

Department of Anesthesiology, Tianjin Fourth Central Hospital, Tianjin City, People's Republic of China.

出版信息

Cancer Manag Res. 2020 Nov 3;12:11145-11155. doi: 10.2147/CMAR.S274975. eCollection 2020.

Abstract

BACKGROUND

Glioma is a heterogeneous aggressive tumor. Ropivacaine, a widely used anesthetic, has been shown to repress the progression of multiple cancers, including glioma. In this study, the effects of ropivacaine on cell proliferation, migration, invasion and apoptosis in glioma were revealed.

METHODS

The expression levels of circSCAF11 and miR-145-5p were detected by quantitative real-time polymerase chain reaction (qRT-PCR) in glioma tissues and cells. The expression levels of epithelial-mesenchymal transition (EMT)-related proteins were determined by Western blot. Oxidative stress was evaluated by the measurement of reactive oxygen species (ROS) and determination of mitochondrial 8-hydroxy-2-deoxyguanosine (8-OHdG) assay in glioma cells. Cell proliferation was determined by cell counting kit-8 (CCK-8) assay and cell colony formation assay. Cell apoptosis and metastasis were detected by flow cytometry analysis and transwell assay, respectively. The binding relationship between circSCAF11 and miR-145-5p was predicted by circular RNA Interactome and identified by dual-luciferase reporter assay and RNA immunoprecipitation assay. In vivo tumor formation assay was performed to reveal the effects between ropivacaine and circSCAF11 overexpression on tumorigenesis in vivo.

RESULTS

CircSCAF11 expression was obviously upregulated and miR-145-5p was significantly downregulated in glioma tissues and cells compared with control groups. Ropivacaine treatment upregulated E-cadherin protein expression and repressed the protein expression of Vimentin. Functionally, ropivacaine exposure promoted ROS and 8-OHdG production and cell apoptosis, whereas inhibited cell proliferation, migration and invasion; however, these effects were hindered by circSCAF11 overexpression. Mechanistically, circSCAF11 was a sponge of miR-145-5p. In addition, ropivacaine was revealed to inhibit tumor growth in vivo by regulating circSCAF11 and miR-145-5p expression.

CONCLUSION

Ropivacaine suppressed glioma progression by regulating circSCAF11 and miR-145-5p, which might provide a theoretical foundation in glioma treatment.

摘要

背景

胶质瘤是一种异质性侵袭性肿瘤。罗哌卡因是一种广泛使用的麻醉剂,已被证明可抑制包括胶质瘤在内的多种癌症的进展。在本研究中,揭示了罗哌卡因对胶质瘤细胞增殖、迁移、侵袭和凋亡的影响。

方法

采用定量实时聚合酶链反应(qRT-PCR)检测胶质瘤组织和细胞中circSCAF11和miR-145-5p的表达水平。通过蛋白质印迹法测定上皮-间质转化(EMT)相关蛋白的表达水平。通过测量胶质瘤细胞中的活性氧(ROS)和测定线粒体8-羟基-2'-脱氧鸟苷(8-OHdG)来评估氧化应激。通过细胞计数试剂盒-8(CCK-8)测定法和细胞集落形成测定法测定细胞增殖。分别通过流式细胞术分析和Transwell测定法检测细胞凋亡和转移。通过环状RNA相互作用组预测circSCAF11与miR-145-5p之间的结合关系,并通过双荧光素酶报告基因测定法和RNA免疫沉淀测定法进行鉴定。进行体内肿瘤形成试验以揭示罗哌卡因与circSCAF11过表达对体内肿瘤发生的影响。

结果

与对照组相比,胶质瘤组织和细胞中circSCAF11表达明显上调,miR-145-5p明显下调。罗哌卡因处理上调E-钙黏蛋白表达并抑制波形蛋白的蛋白表达。在功能上,罗哌卡因暴露促进ROS和8-OHdG产生以及细胞凋亡,而抑制细胞增殖、迁移和侵袭;然而,circSCAF11过表达阻碍了这些作用。机制上,circSCAF11是miR-145-5p的海绵。此外,揭示罗哌卡因通过调节circSCAF11和miR-145-5p表达在体内抑制肿瘤生长。

结论

罗哌卡因通过调节circSCAF11和miR-145-5p抑制胶质瘤进展,这可能为胶质瘤治疗提供理论基础。

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