Gastroenterology, The Second Hospital of Dalian Medical University, No.467, Zhongshan Road, Dalian, Liaoning, China.
Thoracic Surgery, Affiliated Zhongshan Hospital of Dalian University, Dalian, Liaoning, China.
Mol Cell Biochem. 2020 Sep;472(1-2):115-123. doi: 10.1007/s11010-020-03790-y. Epub 2020 Jun 21.
In this study, the functional role of miR-550a-3 and its direct target nuclear factor IC (NFIC) in esophageal squamous cell cancer (ESCC) cells were explored. Differential expression of miR-550a-3 in ESCC tissues was acquired from TCGA database, and Kaplan-Meier method was used to determine the relationship between miR-550a-3 expression and survival time of ESCC patients. Expression level of miR-550a-3 in several ESCC cell lines was measured by qRT-PCR. Two cell lines including Eca109 and JAR were used to perform proliferation, cloning, invasion and migration experiments. Targeted relationship between miR-550a-3 and NFIC was speculated by predication software and confirmed by dual luciferase assay. Additionally, potential relationship between miR-550a-3 and NFIC was analyzed by Spearman rank correlation analysis and western blot. Rescue assays were performed to explore the function of miR-550a-3/NFIC in ESCC cells biological behaviors. Expression levels of key proteins involved in epithelial-to-mesenchymal transition (EMT) process were determined by western blot. By consulting TCGA database, we found that high expression of miR-550a-3 was positively connected with the poor prognosis of patients with ESCC. In addition, overexpression of miR-550a-3 promoted the proliferation, colony formation and metastasis of ESCC cells. Moreover, rescue assays revealed that overexpression of NFIC attenuated the promoting effects of miR-550a-3 on ESCC cells malignant behaviors. While the promoting effects of miR-550a-3 on EMT process were inhibited by NFIC. Our results illustrate the importance of miR-550a-3/NFIC in regulation of ESCC cells growth and metastasis, which could contribute to developing novel target for early diagnosis or neoteric therapeutic target for ESCC.
在这项研究中，我们探讨了 miR-550a-3 及其直接靶标核因子 IC（NFIC）在食管鳞状细胞癌（ESCC）细胞中的功能作用。我们从 TCGA 数据库中获得了 miR-550a-3 在 ESCC 组织中的差异表达，并使用 Kaplan-Meier 方法确定 miR-550a-3 表达与 ESCC 患者生存时间的关系。通过 qRT-PCR 测量了几种 ESCC 细胞系中 miR-550a-3 的表达水平。使用 Eca109 和 JAR 两种细胞系进行增殖、克隆、侵袭和迁移实验。通过预测软件推测 miR-550a-3 和 NFIC 之间的靶向关系，并通过双荧光素酶报告基因实验进行验证。此外，通过 Spearman 秩相关分析和 Western blot 分析了 miR-550a-3 和 NFIC 之间的潜在关系。进行挽救实验以探讨 miR-550a-3/NFIC 在 ESCC 细胞生物学行为中的功能。通过 Western blot 测定 EMT 过程中关键蛋白的表达水平。通过查阅 TCGA 数据库，我们发现 miR-550a-3 的高表达与 ESCC 患者的不良预后呈正相关。此外，miR-550a-3 的过表达促进了 ESCC 细胞的增殖、集落形成和转移。此外，挽救实验表明 NFIC 的过表达减弱了 miR-550a-3 对 ESCC 细胞恶性行为的促进作用。而 miR-550a-3 对 EMT 过程的促进作用则被 NFIC 抑制。我们的研究结果表明，miR-550a-3/NFIC 在调节 ESCC 细胞生长和转移方面具有重要作用，这可能有助于开发 ESCC 的早期诊断新靶点或新型治疗靶点。
