Department of Respiratory Medicine, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong 510120, PR China.
Zhongshan School of Medicine, Sun Yat-sen University-Michigan State University Joint Center of Vector Control for Tropical Diseases, Guangzhou, Guangdong 510080, PR China.
Gene. 2018 Sep 5;670:31-37. doi: 10.1016/j.gene.2018.05.100. Epub 2018 May 24.
MicroRNAs (miRNAs) play important roles in tumorigenesis and tumor progression. In this study, we investigated the role of miR-320a-3p in non-small cell lung cancer (NSCLC). Expressions of miR-320a-3p were firstly determined in 80 NSCLC patients' cancer tissues and adjacent normal lung tissues by qRT-PCR. Then MTT assay, cell migration and invasion assays were performed in vitro. Potential binding sites on target gene of miR-320a-3p were predicted and luciferase reporter assay was used to identify the potential binding sites. Tumorigenesis assay were performed in nude mice by injecting A549 cells which stably express miR-320a-3p. Results indicated that high expression of miR-320a-3p suppresses cell proliferation, migration and invasion through the inactivation of PI3K/Akt signaling pathway in NSCLC cells. Smaller tumor size and lighter weight were also found in nude mice which had miR-320a-3p higher expressed. Furthermore, data from luciferase reporter assay proved the direct binding of miR-320a-3p on the 3'UTR region of ELF3 mRNA, this could further decrease ELF3 expression transcriptionally. We provided evidence that miR-320a-3p might work as a tumor suppressor in NSCLC both in vivo and in vitro.
微小 RNA(miRNAs)在肿瘤发生和肿瘤进展中发挥重要作用。在本研究中,我们研究了 miR-320a-3p 在非小细胞肺癌(NSCLC)中的作用。首先通过 qRT-PCR 测定了 80 例 NSCLC 患者癌组织和相邻正常肺组织中 miR-320a-3p 的表达。然后进行了体外 MTT 测定、细胞迁移和侵袭测定。预测了 miR-320a-3p 靶基因的潜在结合位点,并使用荧光素酶报告基因测定来鉴定潜在的结合位点。通过向稳定表达 miR-320a-3p 的 A549 细胞中注射来在裸鼠中进行肿瘤发生测定。结果表明,miR-320a-3p 的高表达通过失活 PI3K/Akt 信号通路抑制 NSCLC 细胞的增殖、迁移和侵袭。在 miR-320a-3p 表达较高的裸鼠中,还发现肿瘤体积较小,重量较轻。此外,荧光素酶报告基因测定的数据证明了 miR-320a-3p 与 ELF3 mRNA 的 3'UTR 区域的直接结合,这可以进一步降低 ELF3 表达的转录。我们提供了证据表明,miR-320a-3p 可能在体内和体外均作为 NSCLC 的肿瘤抑制因子发挥作用。