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miR-550a-3/NFIC 在食管鳞癌细胞增殖和转移中发挥驱动作用,部分通过 EMT 过程。

miR-550a-3/NFIC plays a driving role in esophageal squamous cell cancer cells proliferation and metastasis partly through EMT process.

机构信息

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.

出版信息

Mol Cell Biochem. 2020 Sep;472(1-2):115-123. doi: 10.1007/s11010-020-03790-y. Epub 2020 Jun 21.

Abstract

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.

摘要

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。

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