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lncRNA CASC2在儿童哮喘血清中的差异表达及其在气道平滑肌细胞增殖和迁移中的作用

Differential Expression of lncRNA CASC2 in the Serum of Childhood Asthma and Its Role in Airway Smooth Muscle Cells Proliferation and Migration.

作者信息

Yang Yane, Sun Zhihong, Ren Tingting, Lei Wei

机构信息

Department of Pediatrics, The Second Affiliated Hospital of Xi'an Medical University, Xi'an, Shaanxi, People's Republic of China.

Department of Pediatrics, Chang 'an District Hospital, Xi'an, Shaanxi, People's Republic of China.

出版信息

J Asthma Allergy. 2022 Feb 11;15:197-207. doi: 10.2147/JAA.S337236. eCollection 2022.

Abstract

PURPOSE

Asthma is a common chronic disease in children. Abnormal expression of lncRNAs can be used as biomarkers for early diagnosis of asthma. The present study aimed to explore the expression change and clinical value of lncRNA CASC2 in asthma, and further investigate its potential mechanism.

PATIENTS AND METHODS

Seventy asthma children and 66 healthy controls were recruited. Levels of mRNAs were detected using qRT-PCR. Receiver operating characteristic (ROC) curves were drawn for diagnostic value evaluation. Asthma cell models were established using PDGF-BB in Human airway smooth muscle cells (ASMCs). Levels of Th1/Th2 related cytokines were detected using ELISA. Lipofectamine 3000 was used for cell transfection. The target relationship was verified using luciferase activity assay.

RESULTS

CASC2 was at a low level in asthma children in comparison with the healthy controls. Serum CASC2 can distinguish healthy individuals from asthma children. Overexpression of CASC2 inhibited PDGF-BB induced cell proliferation and migration. CASC2 upregulation inhibited the release of Th2 related cytokines (IL-4 and IL-10), but promoted the release of Th1-related cytokine (IFN-γ). In PDGF-BB treated ASMCs, the reduced expression of contractile phenotype marker (α-SMA) was detected, but the trend was reversed by CASC2 upregulation. LncRNA CASC2 serves as a ceRNA of miR-31-5p, overexpression of miR-31-5p reversed the influence of CASC2 on asthma in vitro.

CONCLUSION

Serum CASC2 can distinguish healthy individuals from asthma children. CASC2 may be involved in childhood asthma through inhibiting ASMCs proliferation, migration and inflammation via sponging miR-31-5p.

摘要

目的

哮喘是儿童常见的慢性疾病。长链非编码RNA(lncRNA)的异常表达可作为哮喘早期诊断的生物标志物。本研究旨在探讨lncRNA CASC2在哮喘中的表达变化及临床价值,并进一步研究其潜在机制。

患者与方法

招募70例哮喘儿童和66例健康对照。采用qRT-PCR检测mRNA水平。绘制受试者工作特征(ROC)曲线进行诊断价值评估。使用血小板衍生生长因子-BB(PDGF-BB)在人气道平滑肌细胞(ASMCs)中建立哮喘细胞模型。采用酶联免疫吸附测定(ELISA)检测Th1/Th2相关细胞因子水平。使用Lipofectamine 3000进行细胞转染。通过荧光素酶活性测定验证靶标关系。

结果

与健康对照相比,哮喘儿童中CASC2水平较低。血清CASC2可区分健康个体与哮喘儿童。CASC2的过表达抑制了PDGF-BB诱导的细胞增殖和迁移。CASC2上调抑制了Th2相关细胞因子(IL-4和IL-10)的释放,但促进了Th1相关细胞因子(IFN-γ)的释放。在PDGF-BB处理的ASMCs中,检测到收缩表型标志物(α-SMA)的表达降低,但CASC2上调可逆转这一趋势。lncRNA CASC2作为miR-31-5p的竞争性内源RNA(ceRNA),miR-31-5p的过表达逆转了CASC2在体外对哮喘的影响。

结论

血清CASC2可区分健康个体与哮喘儿童。CASC2可能通过海绵化miR-31-5p抑制ASMCs增殖、迁移和炎症反应,从而参与儿童哮喘的发生。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/66e5/8847142/57d97a6f1b70/JAA-15-197-g0001.jpg

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