miR-150-5p Inhibits Hepatoma Cell Migration and Invasion by Targeting MMP14

<div><p>Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related mortality worldwide. Despite progress in diagnostics and treatment of HCC, its prognosis remains poor because the molecular mechanisms underlying hepatocarcinogenesis are not well understood. In the study, we focused on identifying the role of miRNAs in HCC progression. miRNA microarray was used to analyze the differentially expressed miRNAs, and the results were validated by qPCR. We found that the miR-150-5p expression is down-regulated in HCC tissues compared with pair non-tumor tissues. miR-150-5p expression is also decreased in metastatic cancer tissues compared with pair primary tissues, indicating that miR-150-5p may be involved in HCC metastasis. Functionally, miR-150-5p inhibition significantly promotes hepatoma cell migration and invasion, whereas miR-150-5p overexpression suppresses cancer cell migration and invasion <i>in</i><i>vitro</i>. The matrix metalloproteinase 14 (MMP14) is identified as a new target gene of miR-150-5p. miR-150-5p markedly inhibits MMP14 expression in hepatoma cells, and miR-150-5p expression is negative correlation with MMP14 expression <i>in</i><i>vivo</i>. More important, re-expression of MMP14 in hepatoma cells partially reverses the effect of miR-150-5p in inhibiting cell invasion.</p></div

The characteristics of patients with HCC.

<p>Cell invasion assay of Huh7 cells (A) or HepG2 cells (B) after miR-150-5p overexpression or miR-150-5p plus MMP14 overexpression. Data are shown as the mean ± SD based on at least three independent experiments (C). *<i>p</i><0.05.</p><p>The characteristics of patients with HCC.</p

The characteristics of patients with HCC.

<p>Cell invasion assay of Huh7 cells (A) or HepG2 cells (B) after miR-150-5p overexpression or miR-150-5p plus MMP14 overexpression. Data are shown as the mean ± SD based on at least three independent experiments (C). *<i>p</i><0.05.</p><p>The characteristics of patients with HCC.</p

miR-150-5p directly targets MMP14.

<p>(A) Schematic representation of the miR-150-5p site in MMP14 3′-UTR. (B) The 3′UTR reporter assay was carried out in HepG2 cells overexpressed with miR-150-5p. pGL3-MMP14-3′-UTR-WT or pGL3-MMP14-3′-UTR-Mutation was co-transfected with pRL-TK. Luciferase assays were performed 48 h after transfection. Firefly luciferase activity was standardized to Renilla luciferase control. *<i>p</i><0.05. (C and D) Western blot analysis for endogenous MMP14 protein level after miR-150-5p overexpression in hepatoma cells. (E) Western blot analysis for endogenous MMP14 protein level after miR-150-5p inhibition in hepatoma cells. miR-150-5p-inh, miR-150-5p inhibitor. (F) A significant negative correlation between miR-150-5p and MMP14 expression <i>in</i><i>vivo</i> (<i>r</i>² = 0.15389 <i>p</i> = 0.0019).</p

miR-150-5p knockdown promotes hepatoma cell migration and invasion.

<p>(A) Quantitative RT-PCR analysis of miR-150-5p expression after miR-150-5p inhibitor treatment in Huh7 and SMMC 7721 cells. * <i>p</i><0.05. (B and C) Cell migration assay of Huh7 cells (B) or HepG2 cell (C) after miR-150-5p knockdown for 48 h. (D and E) Cell invasion assay of Huh7 cells after miR-150-5p knockdown. Data are shown as the mean ± SD based on at least three independent experiments. *<i>p</i><0.05. (F) Incidence and number of visible metastases per lung in each cohort following subcutaneous inoculation. *<i>p</i><0.05.</p