Novel-miR-4885 Promotes Migration and Invasion of Esophageal Cancer Cells Through Targeting CTNNA2
Novel-miR-4885 Promotes Migration and Invasion of Esophageal Cancer Cells Through Targeting CTNNA2
复制标题
Novel-miR-4885通过靶向CTNNA2促进食管癌细胞的迁移和侵袭
DOI：
10.1089/dna.2018.4377
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发表时间：
2018-12-08
影响因子：
3.1
通讯作者：
Liu, Ran
中科院分区：
文献类型：
--
作者：
Song, Jing;Zhang, Peng;Liu, Ran
食管癌是全世界最常见的癌症之一。找到食管癌的早期诊断生物标志物至关重要。 MicroRNA（miRNA）在食管癌的发生、发展中发挥着重要的调控作用，具有诊断和预后价值。该研究的目的是评估新型 miRNA 的潜在诊断价值。使用 miRDeep2 软件预测新的 miRNA，并通过定量逆转录 PCR (qRT-PCR) 和 PCR 产物的 TA 克隆测序进行验证。还通过 qRT-PCR 分析了食管癌组织和癌旁组织中新型 miRNA 的表达。采用EdU染色和Transwell法检测细胞增殖、迁移、侵袭能力。此外，通过qRT-PCR、蛋白质印迹、荧光素酶报告基因测定和RNA免疫沉淀验证了novel-miR-4885的靶基因CTNNA2。我们鉴定了新型-miR-4885，并通过qRT-PCR证实了食管癌细胞中的表达水平。 TA克隆测序结果与预测一致，pre-miRNA具有标准的发夹茎环结构。此外，与癌旁组织相比，食管癌组织中novel-miR-4885的表达上调（p < 0.05）。此外，分析表明，过表达novel-miR-4885可以改善细胞增殖、迁移和侵袭，平均倍数变化分别为1.19、1.59和2.34。 Novel-miR-4885可以与CTNNA2的3'非翻译区结合，减少细胞粘附并促进食管癌细胞的上皮间质转化。通过蛋白质印迹，N-钙粘蛋白、β-连环蛋白、波形蛋白和α-平滑肌肌动蛋白的表达上调，而E-钙粘蛋白和ZO-1蛋白的表达下调。食管癌细胞中存在的新 miRNA 得到验证，补充了 miRNA 数据库。同时探讨了其可能的功能机制，结果表明新的miRNA可能作为食管癌诊断的潜在生物标志物。
Esophageal cancer is one of the most common cancers worldwide. It is critical to find early diagnostic biomarkers for esophageal cancer. MicroRNAs (miRNAs) play important regulatory roles in occurrence and development of esophageal cancer, which has the diagnostic and prognostic values. The aim of the study was to evaluate the potential diagnostic value of the novel miRNA. The novel miRNA was predicted using miRDeep2 software and validated by quantitative reverse transcription PCR (qRT-PCR) and the TA-cloning sequencing of the PCR products. The expression of the novel miRNA in esophageal cancer tissues and adjacent tissues was also analyzed by qRT-PCR. The EdU staining and transwell method were used to detect the capacity of cell proliferation, migration, and invasion. Besides, the target gene CTNNA2 of novel-miR-4885 was verified via qRT-PCR, western blot, luciferase reporter assay, and RNA immunoprecipitation. We identified the novel-miR-4885, the expression level was confirmed by qRT-PCR in the esophageal cancer cells. The result of TA-cloning sequencing was consistent with the prediction and the pre-miRNA had a standard hairpin stem-loop structure. In addition, the expression of novel-miR-4885 was upregulated in esophageal cancer tissue compared with that in adjacent tissue (p < 0.05). Further, the assays showed that overexpression novel-miR-4885 could improve the cell proliferation, migration, and invasion with an average fold change of 1.19, 1.59, and 2.34, respectively. Novel-miR-4885 can bind to 3 ' untranslated region of CTNNA2 to reduce cell adhesion and promote epithelial-mesenchymal transition in esophageal cancer cell. The expression of N-cadherin, beta-catenin, Vimentin, and alpha-smooth muscle actin was upregulated, while that of E-cadherin and ZO-1 proteins was downregulated through western blot. Novel miRNA present in esophageal cancer cells was validated, supplementing the miRNA database. Meantime, the possible functional mechanisms were explored, and the results showed that the novel miRNA may serve as potential biomarker for the diagnosis of esophageal cancer.