Shi Jiang, Wang Huan, Feng Wanlu, Huang Siyuan, An Jinlu, Wang Lifang, Jiang Junguang
Department of Respiratory, The First Affiliated Hospital of Zhengzhou University, No.1 Jianshe East Road, Zhengzhou, 450052, Henan, China.
Department of Oncology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan, China.
Hum Cell. 2021 Sep;34(5):1490-1503. doi: 10.1007/s13577-021-00573-5. Epub 2021 Jul 6.
Circular RNAs (circRNAs) play a significant role in the progression of diverse malignancies. Here, we aimed to probe the function and mechanism of circ_0069244 in non-small cell lung cancer (NSCLC). In the present study, circ_0069244 was selected from the circRNA microarray datasets (GSE112214). Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to examine circ_0069244, miR-346 and XPC complex subunit, DNA damage recognition and repair factor (XPC) expression levels. Kaplan-Meier curve was employed to analyze the association between circ_0069244 expression and overall survival of NSCLC patients. Cell counting kit-8 (CCK-8) and 5-Bromo-2'-deoxyuridine (BrdU) experiments were utilized to examine the proliferation of NSCLC cells. Scratch healing and Transwell experiments were executed to examine the migration of NSCLC cells. Western blot was conducted to detect XPC expression at protein level in NSCLC cells. Bioinformatics analysis, dual-luciferase reporter gene and RNA immunoprecipitation (RIP) experiments predicted and validated the targeting relationships of circ_0069244 and miR-346, as well as miR-346 and 3'untranslated region (UTR) of XPC mRNA, respectively. We reported that circ_0069244 was remarkably down modulated in NSCLC and was linked to shorter survival and poor tumor histological grade in NSCLC patients. Functionally, circ_0069244 repressed NSCLC cell proliferation and migration. Furthermore, miR-346-5p was unveiled to be a downstream target of circ_0069244, and miR-346-5p specifically modulated XPC expression. Rescue experiments indicated that the inhibitory effect of circ_0069244 was abolished by co-expression of miR-346-5p mimics. Taken together, circ_0069244 restrained NSCLC progression by modulating the miR-346-5p/XPC axis.
环状RNA（circRNAs）在多种恶性肿瘤的进展中发挥着重要作用。在此，我们旨在探究circ_0069244在非小细胞肺癌（NSCLC）中的功能及机制。在本研究中，circ_0069244是从环状RNA微阵列数据集（GSE112214）中筛选出来的。采用定量实时聚合酶链反应（qRT-PCR）检测circ_0069244、miR-346以及XPC复合物亚基、DNA损伤识别与修复因子（XPC）的表达水平。运用Kaplan-Meier曲线分析circ_0069244表达与NSCLC患者总生存期之间的关联。利用细胞计数试剂盒-8（CCK-8）和5-溴-2'-脱氧尿苷（BrdU）实验检测NSCLC细胞的增殖情况。进行划痕愈合实验和Transwell实验检测NSCLC细胞的迁移情况。通过蛋白质印迹法检测NSCLC细胞中XPC蛋白水平的表达。生物信息学分析、双荧光素酶报告基因实验以及RNA免疫沉淀（RIP）实验分别预测并验证了circ_0069244与miR-346以及miR-346与XPC mRNA 3'非翻译区（UTR）之间的靶向关系。我们发现circ_0069244在NSCLC中显著下调，且与NSCLC患者较短的生存期及不良的肿瘤组织学分级相关。在功能上，circ_0069244抑制NSCLC细胞的增殖和迁移。此外，miR-346-5p被揭示为circ_0069244的下游靶点，且miR-346-5p特异性调节XPC的表达。挽救实验表明，共表达miR-346-5p模拟物可消除circ_0069244的抑制作用。综上所述，circ_0069244通过调节miR-346-5p/XPC轴抑制NSCLC的进展。
