Ding Wei, Shi Ying, Zhang Hong
Department of Thyroid Surgery, The Second Hospital of Jilin University Changchun, Jilin Province, China.
Am J Transl Res. 2021 Jun 15;13(6):5911-5927. eCollection 2021.
Circular RNAs (circRNA) are found to be closely associated with cancers as their possibility as "sponges" to miRNAs, thus liberating the downstream target mRNA. However, deep research is still needed to study the function of circRNA in papillary thyroid carcinoma (PTC). Here, we sought to explore new circRNA which could play an important role in the development of PTC. We filtered candidate circRNAs based on microarray data from public database and verified the result using qRT-PCR. We performed CCK8 assay, colony formation assay, apoptosis assay, transwell assays, and xenograft experiments to explore the function of selected circRNA on PTC. We predicted the miRNA targets of the circRNA and the target gene of miRNA through bioinformatic analysis and validated the target by mutant experiments. And by the use of overexpression experiments, knockdown experiments, and the functional assays mentioned above, we figured out the pathway behind the selected circRNA. Based on our data, we found that circNEURL4 was significantly decreased in the PTC samples and lower expression of circNEURL4 was closely associated with a poor prognosis of patients with PTC. Then, we proved that circNEURL4 could inhibit cell proliferation and invasion of PTC in vivo and in vitro. Furthermore, we demonstrated that circNEURL4 may binding to miR-1278 and thus indirectly improving the expression of LATS1. Our findings revealed that circNEURL4 may probably serve as a diagnostic marker to predict PTC patients' prognosis and a possible therapeutic target to PTC via miR-1278/LATS1 axis.
环状RNA(circRNA)被发现与癌症密切相关,因为它们有可能作为微小RNA(miRNA)的“海绵”,从而释放下游靶标信使核糖核酸(mRNA)。然而,仍需要深入研究来探究circRNA在甲状腺乳头状癌(PTC)中的功能。在此,我们试图探索在PTC发生发展中起重要作用的新circRNA。我们基于公共数据库的微阵列数据筛选候选circRNA,并使用实时定量逆转录聚合酶链反应(qRT-PCR)验证结果。我们进行了细胞计数试剂盒-8(CCK8)检测、集落形成检测、凋亡检测、Transwell检测和异种移植实验,以探究所选circRNA对PTC的功能。我们通过生物信息学分析预测circRNA的miRNA靶标以及miRNA的靶基因,并通过突变实验验证靶标。通过使用过表达实验、敲低实验以及上述功能检测,我们弄清楚了所选circRNA背后的信号通路。基于我们的数据,我们发现circNEURL4在PTC样本中显著降低,且circNEURL4的低表达与PTC患者的不良预后密切相关。然后,我们证明circNEURL4在体内和体外均可抑制PTC的细胞增殖和侵袭。此外,我们证明circNEURL4可能与miR-1278结合,从而间接提高大肿瘤抑制激酶1(LATS1)的表达。我们的研究结果表明,circNEURL4可能作为预测PTC患者预后的诊断标志物,并可能通过miR-1278/LATS1轴成为PTC的潜在治疗靶点。