Department of Gastroenterology and Hepatology, University Hospital Essen, Essen, Germany.
Mol Cancer. 2010 Aug 27;9:227. doi: 10.1186/1476-4598-9-227.
Micro-ribonucleic acid (miRNA)-199a-5p has been reported to be decreased in hepatocellular carcinoma (HCC) compared to normal tissue. Discoidin domain receptor-1 (DDR1) tyrosine kinase, involved in cell invasion-related signaling pathway, was predicted to be a potential target of miR-199a-5p by the use of miRNA target prediction algorithms. The aim of this study was to investigate the role of miR-199a-5p and DDR1 in HCC invasion.
Mature miR-199a-5p and DDR1 expression were evaluated in tumor and adjacent non-tumor liver tissues from 23 patients with HCC undergoing liver resection and five hepatoma cell lines by the use of real-time quantitative RT-PCR (qRT-PCR) analysis. The effect of aberrant miR-199a-5p expression on cell invasion was assessed in vitro using HepG2 and SNU-182 hepatoma cell lines. Luciferase reporter assay was employed to validate DDR1 as a putative miR-199a-5p target gene. Regulation of DDR1 expression by miR-199a-5p was assessed by the use qRT-PCR and western blotting analysis.
A significant down-regulation of miR-199a-5p was observed in 65.2% of HCC tissues and in four of five cell lines. In contrast, DDR1 expression was significantly increased in 52.2% of HCC samples and in two of five cell lines. Increased DDR1 expression in HCC was associated with advanced tumor stage. DDR1 was shown to be a direct target of miR-199a-5p by luciferase reporter assay. Transfection of miR-199a-5p inhibited invasion of HepG2 but not SNU-182 hepatoma cells.
Decreased expression of miR-199a-5p contributes to increased cell invasion by functional deregulation of DDR1 activity in HCC. However, the effect of miR-199a-5p on DDR1 varies among individuals and hepatoma cell lines. These findings may have significant translational relevance for development of new targeted therapies as well as prognostic prediction for patients with HCC.
与正常组织相比，微小 RNA-199a-5p（miRNA-199a-5p）在肝细胞癌（HCC）中表达降低。通过使用 miRNA 靶标预测算法，发现 discoidin 结构域受体-1（DDR1）酪氨酸激酶参与细胞侵袭相关信号通路，可能是 miR-199a-5p 的潜在靶标。本研究旨在探讨 miR-199a-5p 和 DDR1 在 HCC 侵袭中的作用。
采用实时定量 RT-PCR（qRT-PCR）分析 23 例接受肝切除术的 HCC 患者肿瘤和相邻非肿瘤肝组织以及 5 种肝癌细胞系中成熟 miR-199a-5p 和 DDR1 的表达。使用 HepG2 和 SNU-182 肝癌细胞系评估异常 miR-199a-5p 表达对细胞侵袭的影响。荧光素酶报告实验验证 DDR1 是 miR-199a-5p 的潜在靶基因。采用 qRT-PCR 和 Western blot 分析评估 miR-199a-5p 对 DDR1 表达的调控。
在 65.2%的 HCC 组织和 5 种细胞系中的 4 种中观察到 miR-199a-5p 的显著下调。相反，在 52.2%的 HCC 样本和 5 种细胞系中的 2 种中，DDR1 表达显著增加。HCC 中 DDR1 的增加与肿瘤分期较晚有关。荧光素酶报告实验表明 DDR1 是 miR-199a-5p 的直接靶基因。miR-199a-5p 的转染抑制了 HepG2 但不抑制 SNU-182 肝癌细胞的侵袭。
miR-199a-5p 的表达降低通过功能性下调 DDR1 活性导致 HCC 细胞侵袭增加。然而，miR-199a-5p 对 DDR1 的影响在个体和肝癌细胞系之间存在差异。这些发现可能对开发新的靶向治疗方法以及预测 HCC 患者的预后具有重要的转化意义。
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