Department of Clinical Laboratory, Qilu Hospital of Shandong University, Jinan, China.
Department of Clinical Laboratory, The Second Hospital of Shandong University, Jinan, China.
Thorac Cancer. 2020 Sep;11(9):2506-2516. doi: 10.1111/1759-7714.13562. Epub 2020 Jul 9.
We have previously developed a unique metastasis-associated signature consisting of six long non-coding RNAs (lncRNAs), including a novel lncRNA, namely LINC02323. In the present study, we aimed to investigate the underlying roles of LINC02323 in the migration, invasion and TGF-β-induced epithelial-mesenchymal transition (EMT) of lung adenocarcinoma (LUAD) cells.
The distribution of LINC02323 was detected by the nuclear-plasma separation experiment. Cell proliferation was assessd by MTT assay, and cell migration and invation were detected by transwell assays. EMT was detected by RT-qPCR and western blotting. Interaction between miRNA and LINC02323 was predicted by starBase v2.0 and confirmed by the double luciferase reporting system.
LINC02323 was distributed in the cytoplasm and nucleus. The overexpression or deletion of LINC02323 did not affect the proliferation of LUAD cells, while significantly affected the migration and invasion of LUAD cells. TGF-β-induced EMT process was significantly affected by both RNA interference (RNAi) and overexpression of LINC02323. The predicted results showed that there were binding sites between LINC02323 and miR-1343-3p. The expression of LINC02323 was found to be negatively correlated with miR-1343-3p in LUAD by analyzing The Cancer Genome Atlas (TCGA) database. The double luciferase reporting system, RT-qPCR and western blotting experiments confirmed that LINC02323 could bind to miR-1343-3p, which bound to TGF-β receptor 1 (TGFBR1). Inhibition of miR-1343-3p reversed LINC02323 silencing-mediated suppression of migration, invasion and EMT.
LINC02323 acts as a competing endogenous RNA (ceRNA), which sponged miR-1343-3p to upregulate the TGFBR1 expression and promote the EMT and metastasis in LUAD.
SIGNIFICANT FINDINGS OF THE STUDY: LINC02323 promotes epithelial-mesenchymal transition and metastasis via sponging miR-1343-3p in lung adenocarcinoma.
LINC02323 is a key molecule in the process of invasion and metastasis of LUAD and might be used as a potential target in metastatic cancer.
我们之前开发了一个由六个长非编码 RNA（lncRNA）组成的独特转移相关特征，其中包括一个新型 lncRNA，即 LINC02323。在本研究中，我们旨在研究 LINC02323 在肺腺癌（LUAD）细胞迁移、侵袭和 TGF-β 诱导的上皮-间充质转化（EMT）中的潜在作用。
通过核-浆分离实验检测 LINC02323 的分布。通过 MTT 测定法评估细胞增殖，通过 Transwell 测定法检测细胞迁移和侵袭。通过 RT-qPCR 和 Western blot 检测 EMT。通过 starBase v2.0 预测 miRNA 与 LINC02323 之间的相互作用，并通过双荧光素酶报告系统进行验证。
LINC02323 分布在细胞质和细胞核中。LINC02323 的过表达或缺失均不影响 LUAD 细胞的增殖，但显著影响 LUAD 细胞的迁移和侵袭。RNA 干扰（RNAi）和 LINC02323 的过表达均显著影响 TGF-β 诱导的 EMT 过程。预测结果表明，LINC02323 与 miR-1343-3p 之间存在结合位点。通过分析癌症基因组图谱（TCGA）数据库，发现 LUAD 中 LINC02323 的表达与 miR-1343-3p 呈负相关。双荧光素酶报告系统、RT-qPCR 和 Western blot 实验证实，LINC02323 可与 miR-1343-3p 结合，miR-1343-3p 结合 TGF-β 受体 1（TGFBR1）。抑制 miR-1343-3p 可逆转 LINC02323 沉默介导的迁移、侵袭和 EMT 抑制作用。
LINC02323 作为竞争性内源 RNA（ceRNA），通过海绵吸附 miR-1343-3p 上调 TGFBR1 表达，促进 LUAD 的 EMT 和转移。
研究的重要发现：LINC02323 通过海绵吸附 miR-1343-3p 促进肺腺癌上皮-间充质转化和转移。
LINC02323 是 LUAD 侵袭和转移过程中的关键分子，可能作为转移性癌症的潜在靶点。
