Ren Tiejun, Liu Chang, Hou Jianfeng, Shan Fengxiao
Department of Oncology, Luoyang Central Hospital Affiliated to Zhengzhou University, Luoyang, People's Republic of China.
Onco Targets Ther. 2020 May 7;13:3867-3880. doi: 10.2147/OTT.S235231. eCollection 2020.
Non-small cell lung cancer (NSCLC) is the largest type of lung cancer (LC) with a higher mortality rate. Circular RNAs (circRNAs) have been shown to play an important role in cancer progression. Therefore, this study was to explore the function of hsa_circ_0043265 in NSCLC.
The expression levels of hsa_circ_0043265, microRNA-25-3p (miR-25-3p) and forkhead box P2 (FOXP2) were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Ribonuclease R (RNase R) and Actinomycin D (ActD) were used to verify the authenticity and stability of hsa_circ_0043265. Cell counting kit-8 (CCK-8), flow cytometry and transwell assays were used to evaluate the abilities of proliferation, apoptosis, migration and invasion of NSCLC cells. Also, Western blot (WB) analysis was performed to assess the levels of apoptosis, epithelial-mesenchymal transition (EMT) and proliferation-related proteins and FOXP2 protein. RNA immunoprecipitation (RIP) and dual-luciferase reporter assays were used to verify the interaction between miR-25-3p and hsa_circ_0043265 or FOXP2. Besides, mice xenograft models were constructed to confirm the effect of hsa_circ_0043265 on NSCLC tumor growth in vivo.
Hsa_circ_0043265 was lowly expressed in NSCLC tissues and cells, and its overexpression inhibited the proliferation, migration, invasion and EMT process, while improved the apoptosis of NSCLC cells. MiR-25-3p could be sponged by hsa_circ_0043265, and its overexpression could invert the suppression effect of overexpressed-hsa_circ_0043265 on NSCLC progression. Moreover, FOXP2 was a target of miR-25-3p, and its silencing also could reverse the inhibition effect of overexpressed-hsa_circ_0043265 on NSCLC progression. In addition, hsa_circ_0043265 overexpression reduced the tumor growth of NSCLC in vivo.
Hsa_circ_0043265 could sponge miR-25-3p to improve FOXP2 expression, thereby inhibiting NSCLC progression. This study showed that hsa_circ_0043265 could be a potential biomarker for early diagnosis of NSCLC.
非小细胞肺癌（NSCLC）是肺癌（LC）中最常见的类型，死亡率较高。环状RNA（circRNAs）已被证明在癌症进展中起重要作用。因此，本研究旨在探讨hsa_circ_0043265在NSCLC中的功能。
采用定量实时聚合酶链反应（qRT-PCR）检测hsa_circ_0043265、微小RNA-25-3p（miR-25-3p）和叉头框P2（FOXP2）的表达水平。用核糖核酸酶R（RNase R）和放线菌素D（ActD）验证hsa_circ_0043265的真实性和稳定性。使用细胞计数试剂盒-8（CCK-8）、流式细胞术和Transwell实验评估NSCLC细胞的增殖、凋亡、迁移和侵袭能力。此外，进行蛋白质免疫印迹（WB）分析以评估凋亡、上皮-间质转化（EMT）和增殖相关蛋白以及FOXP2蛋白的水平。采用RNA免疫沉淀（RIP）和双荧光素酶报告基因实验验证miR-25-3p与hsa_circ_0043265或FOXP2之间的相互作用。此外，构建小鼠异种移植模型以证实hsa_circ_0043265对NSCLC肿瘤体内生长的影响。
hsa_circ_0043265在NSCLC组织和细胞中低表达，其过表达抑制了NSCLC细胞的增殖、迁移、侵袭和EMT过程，同时促进了细胞凋亡。hsa_circ_0043265可以吸附miR-25-3p，miR-25-3p过表达可逆转过表达hsa_circ_0043265对NSCLC进展的抑制作用。此外，FOXP2是miR-25-3p的靶标，其沉默也可以逆转过表达hsa_circ_0043265对NSCLC进展的抑制作用。此外，hsa_circ_0043265过表达降低了NSCLC在体内的肿瘤生长。
hsa_circ_0043265可以吸附miR-25-3p以提高FOXP2表达，从而抑制NSCLC进展。本研究表明hsa_circ_0043265可能是NSCLC早期诊断的潜在生物标志物。
