microRNA-126-5p 通过靶向 EGFR 抑制肝癌细胞的增殖、侵袭和迁移。

MicroRNA-126-5p suppresses cell proliferation, invasion and migration by targeting EGFR in liver cancer.

机构信息

The First Affiliated Hospital of Zhejiang University of Traditional Chinese Medicine, NO.1 Building, Yunbin Bay, Jianggan District, Hangzhou 310000, Zhejiang, China.

出版信息

Clin Res Hepatol Gastroenterol. 2020 Nov;44(6):865-873. doi: 10.1016/j.clinre.2020.03.025. Epub 2020 May 12.

DOI:10.1016/j.clinre.2020.03.025

PMID:32409283

Abstract

BACKGROUND

The expression status and potential mechanistic involvement of microRNA (miR)-126-5p in hepatocellular carcinoma (HCC) are still elusive currently. Here we set out to address this issue both in cell lines and in patients' tissues.

METHODS

The relative expression levels of endogenous miR-126-5p and epidermal growth factor receptor (EGFR) were quantified by real-time polymerase chain reaction. Cell viability and proliferation were measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and clonogenic assays, respectively. Cell invasive and migrative capacities were determined by transwell and wound healing assays, respectively. The regulatory action of miR-126-5p on EGFR was interrogated by luciferase reporter assay. Translational level of EGFR was analyzed by Western blotting.

RESULTS

MiR-126-5p was significantly down-regulated in both HCC patients' tissues and cell lines. Forced expression of miR-126-5p greatly compromised cell viability, proliferation, invasion and migration, while miR-126-5p-specific inhibitor promoted these oncogenic phenotypes. MiR-126-5p mimics inhibited endogenous expression of EGFR and suppressed EGFR 3'-untranslated region-fused luciferase activity. Co-expression of EGFR in miR-126-5p-proficient cells completely restored cell migrative and invasive capacities, while co-transfection with EGFR siRNA significantly inhibited miR-126-5p inhibitor-induced cell invasion and migration.

CONCLUSION

MiR-126-5p was aberrantly decreased in HCC and subsequently relieved the suppression on EGFR expression, which consequently contributed to the tumor biology of HCC.

摘要

背景

目前，miR-126-5p 在肝细胞癌（HCC）中的表达状态及其潜在的机制尚不清楚。在这里，我们旨在通过细胞系和患者组织来解决这个问题。

方法

通过实时聚合酶链反应定量检测内源性 miR-126-5p 和表皮生长因子受体（EGFR）的相对表达水平。通过 3-(4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐和集落形成测定法分别测量细胞活力和增殖。通过 Transwell 和划痕愈合测定法分别测定细胞侵袭和迁移能力。通过荧光素酶报告基因测定法检测 miR-126-5p 对 EGFR 的调节作用。通过 Western blot 分析 EGFR 的翻译水平。

结果

miR-126-5p 在 HCC 患者组织和细胞系中均显著下调。miR-126-5p 的强制表达大大降低了细胞活力、增殖、侵袭和迁移能力，而 miR-126-5p 特异性抑制剂则促进了这些致癌表型。miR-126-5p 模拟物抑制内源性 EGFR 的表达，并抑制 EGFR 3'-非翻译区融合荧光素酶活性。在 miR-126-5p 丰富的细胞中共表达 EGFR 完全恢复了细胞迁移和侵袭能力，而共转染 EGFR siRNA 则显著抑制了 miR-126-5p 抑制剂诱导的细胞侵袭和迁移。