Li Na, Zhu Dan
Department of Respiratory, The Second Affiliated Hospital of Dalian Medical University, Dalian, China.
Cell Cycle. 2021 May 4:1-13. doi: 10.1080/15384101.2021.1919829.
Circular RNAs (circRNAs) have been reported to exert vital roles in the tumorigenesis of non-small cell lung cancer (NSCLC). The study aimed to probe the function of circ_0017956 in NSCLC development. The expression of circ_0017956, microRNA (miR)-515-5p and integrin subunit beta 8 (ITGB8) was gauged by quantitative real-time polymerase chain reaction (qRT-PCR) or western blot. The proliferation detection was conducted employing Cell Counting Kit-8 (CCK-8) and colony formation assays. Transwell assay was performed to determine cell migratory and invasive abilities. Western blot was implemented for the measurement of related proteins. The targeted interactions among circ_0017956, miR-515-5p and ITGB8 were affirmed by dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay and RNA pull-down assay. The role of circ_0017956 in NSCLC tumor growth was studied by xenograft mice model. Circ_0017956 and ITGB8 abundances were overtly raised whereas miR-515-5p was low expressed in NSCLC tissues and cells. Circ_0017956 knockdown caused inhibitory effects on the proliferative and metastasizing capacities of NSCLC cells. Mechanistically, circ_0017956 could sponge miR-515-5p, and circ_0017956 depletion blocked NSCLC cell malignant behaviors by increasing miR-515-5p expression. Furthermore, miR-515-5p targeted ITGB8 and ITGB8 overexpression also neutralized the miR-515-5p-triggered inhibition effects on NSCLC cell progression. Moreover, circ_0017956 could regulate ITGB8 expression through sponging miR-515-5p. In addition, circ_0017956 knockdown repressed NSCLC tumorigenesis by targeting miR-515-5p/ITGB8 axis . Circ_0017956 could promote NSCLC carcinogenesis at least partly through sponging miR-515-5p and upregulating ITGB8 level, providing a novel theoretical basis for NSCLC treatment.
据报道，环状RNA（circRNAs）在非小细胞肺癌（NSCLC）的肿瘤发生中发挥着重要作用。本研究旨在探究circ_0017956在NSCLC发展中的功能。通过定量实时聚合酶链反应（qRT-PCR）或蛋白质免疫印迹法检测circ_0017956、微小RNA（miR）-515-5p和整合素β8亚基（ITGB8）的表达。采用细胞计数试剂盒-8（CCK-8）和集落形成试验进行增殖检测。进行Transwell试验以确定细胞的迁移和侵袭能力。采用蛋白质免疫印迹法检测相关蛋白。通过双荧光素酶报告基因试验、RNA免疫沉淀（RIP）试验和RNA下拉试验确定circ_0017956、miR-515-5p和ITGB8之间的靶向相互作用。通过异种移植小鼠模型研究circ_0017956在NSCLC肿瘤生长中的作用。在NSCLC组织和细胞中，circ_0017956和ITGB8的丰度明显升高，而miR-515-5p表达较低。敲低circ_0017956对NSCLC细胞的增殖和转移能力产生抑制作用。机制上，circ_0017956可以吸附miR-515-5p，circ_0017956缺失通过增加miR-515-5p表达来阻断NSCLC细胞的恶性行为。此外，miR-515-5p靶向ITGB8，ITGB8过表达也可抵消miR-515-5p对NSCLC细胞进展的抑制作用。此外，circ_0017956可以通过吸附miR-515-5p来调节ITGB8的表达。此外，敲低circ_0017956通过靶向miR-515-5p/ITGB8轴抑制NSCLC肿瘤发生。circ_0017956至少部分通过吸附miR-515-5p和上调ITGB8水平促进NSCLC癌变，为NSCLC治疗提供了新的理论依据。
