项保利*, 苏菁, 刘洋
许多研究表明环状RNA (circular RNAs, circRNAs)在肿瘤的整个生物过程中发挥着关键的调控作用。本研究旨在探讨circ_0001666在非小细胞肺癌(non-small cell lung cancer, NSCLC)细胞中的生物学功能及其分子机制，以期为NSCLC的早期诊断和治疗提供新的靶点。在公共基因芯片数据库GEO (Gene Expression Omnibus )中下载GSE101586数据集，通过GEO2R分析得到差异基因。采用实时定量聚合酶链反应检测NSCLC细胞中circ_0001666的表达水平。采用细胞计数试剂盒-8和细胞凋亡检测试剂盒分别检测敲低circ_0001666对NSCLC细胞增殖和凋亡的作用。通过生物信息学预测、双荧光素酶报告实验、RNA结合蛋白免疫沉淀实验和RNA pull down实验研究微小RNA 330-5p (microRNA 330-5p, miR-330-5p)与circ_0001666、高迁移率蛋白A2 (high mobility group A2 protein, HMGA2)之间的靶向关系。结果显示：circ_0001666在NSCLC细胞系中的表达显著高于正常肺上皮细胞。敲低circ_0001666显著降低了NSCLC细胞的增殖活力，并促进了细胞凋亡，而转染miR-330-5p抑制剂可逆转此作用。miR-330-5p是circ_0001666的下游靶点，可以被circ_0001666吸附；HMGA2是miR-330-5p的靶基因，可以被circ_0001666间接正向调控。以上结果提示，circ_0001666通过靶向调控miR-330-5p/HMGA2轴促进NSCLC细胞的增殖并抑制凋亡。
rc_0001666 promotes the proliferation and inhibits apoptosis of non-small cell lung cancer cells by targeting the miR-330-5p/HMGA2 axis
XIANG Bao-Li*, SU Jing, LIU Yang
Department of Respiratory and Critical Care Medicine, the First Affiliated Hospital of Hebei North University, Zhangjiakou 075000, China
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.
通讯作者：项保利 E-mail: email@example.com
项保利, 苏菁, 刘洋. Circ_0001666通过靶向miR-330-5p/HMGA2轴促进非小细胞肺癌细胞的增殖并抑制凋亡[J]. 生理学报 2021; 73 (3): 491-500.
XIANG Bao-Li, SU Jing, LIU Yang. rc_0001666 promotes the proliferation and inhibits apoptosis of non-small cell lung cancer cells by targeting the miR-330-5p/HMGA2 axis. Acta Physiol Sin 2021; 73 (3): 491-500 (in Chinese with English abstract).