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[环状RNA_0001666通过靶向miR-330-5p/HMGA2轴促进非小细胞肺癌细胞增殖并抑制其凋亡]

[Circ_0001666 promotes the proliferation and inhibits apoptosis of non-small cell lung cancer cells by targeting the miR-330-5p/HMGA2 axis].

作者信息

Xiang Bao-Li, Su Jing, Liu Yang

机构信息

Department of Respiratory and Critical Care Medicine, the First Affiliated Hospital of Hebei North University, Zhangjiakou 075000, China.

出版信息

Sheng Li Xue Bao. 2021 Jun 25;73(3):491-500.

Abstract

Many studies have shown that circular RNAs (circRNAs) play a key regulatory role in the whole biological process of tumors. The purpose of this study was to explore the biological function and molecular mechanism of circ_0001666 in non-small cell lung cancer (NSCLC), so as to provide new targets for the diagnosis and treatment of NSCLC. Gene expression profiles were downloaded from Gene Expression Omnibus (GEO, GSE101586) and the differential genes were obtained by using GEO2R analysis. The quantitative real time polymerase chain reaction (qRT-PCR) was used to detect the expression level of circ_0001666 in NSCLC cells. Cell counting kit-8 (CCK-8) and Annexin V-FITC apoptosis detection kit were respectively used to assess the cell proliferation and apoptosis, where circ_0001666 was knockdown in NSCLC cells. The targeted relationship among mircoRNA 330-5p (miR-330-5p), circ_0001666, and high mobility group A2 protein (HMGA2) was verified by bioinformatics prediction, dual-luciferase reporter gene, RNA immunoprecipitation (RIP) and RNA pull down assay. The results showed that the expression of circ_0001666 in NSCLC cells was significantly up-regulated than that in normal lung epithelial cells. Circ_0001666 knockdown reduced the cell viability and promoted the apoptosis of NSCLC cells, which could be reversed by miR-330-5p inhibitors. MiR-330-5p is the downstream target of circ_0001666 and can be adsorbed by circ_0001666. HMGA2 is a target gene of miR-330-5p, which can be indirectly regulated by circ_0001666. The results suggest that circ_0001666 promotes the proliferation and inhibits apoptosis of NSCLC cells via miR-330-5p/HMGA2 axis.

摘要

许多研究表明,环状RNA(circRNAs)在肿瘤的整个生物学过程中发挥着关键的调控作用。本研究的目的是探讨circ_0001666在非小细胞肺癌(NSCLC)中的生物学功能和分子机制,从而为NSCLC的诊断和治疗提供新的靶点。从基因表达综合数据库(GEO,GSE101586)下载基因表达谱,并使用GEO2R分析获得差异基因。采用定量实时聚合酶链反应(qRT-PCR)检测circ_0001666在NSCLC细胞中的表达水平。分别使用细胞计数试剂盒-8(CCK-8)和膜联蛋白V-FITC凋亡检测试剂盒评估NSCLC细胞中circ_0001666敲低后的细胞增殖和凋亡情况。通过生物信息学预测、双荧光素酶报告基因、RNA免疫沉淀(RIP)和RNA下拉实验验证了微小RNA 330-5p(miR-330-5p)、circ_0001666和高迁移率族蛋白A2(HMGA2)之间的靶向关系。结果显示,circ_0001666在NSCLC细胞中的表达明显高于正常肺上皮细胞。敲低circ_0001666可降低NSCLC细胞的活力并促进其凋亡,而miR-330-5p抑制剂可逆转这种作用。miR-330-5p是circ_0001666的下游靶点,可被circ_0001666吸附。HMGA2是miR-330-5p的靶基因,可被circ_0001666间接调控。结果表明,circ_0001666通过miR-330-5p/HMGA2轴促进NSCLC细胞的增殖并抑制其凋亡。

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