Aims

Angelol-A (Ang-A), a kind of coumarins, is isolated from the roots of Angelica pubescens f. biserrata. However, AA exerts antitumor effects and molecular mechanism on cervical cancer cells is unknown.

Main methods

Cell viability was determined using the MTT assay, and the cell cycle phase was assessed by PI staining with flow cytometry. Ang-A-treated cells with/without Antago-miR-29a-3p (miR-29a-3p inhibitor) or U0126 (MEK inhibitor) were assessed for the expression of miR-29a-3p, in vitro migration/invasion, and angiogenesis using qRT-PCR, a chemotaxis assay, and tube formation assay, respectively. The expression of mitogen-activated protein kinases/MMP2/MMP9/VEGFA was determined by western blot analysis with applicable antibodies.

Key findings

Ang-A significantly inhibited MMP2 and VEGFA expression, cell migration, and invasive motility in human cervical cancer cells. Conditioned medium inhibited tube formation in HUVECs. Ang-A principally inhibited invasive motility and angiogenesis by upregulating the expression of miR-29a-3p that targets the VEGFA-3′ UTR. The role of miR-29a-3p was confirmed using Antago-miR-29a-3p, which reversed the Ang-A-inhibited expression of MMP2 and VEGFA, invasive motility, and angiogenesis in human cervical cancer cells. The ERK pathway was implicated in mediating the metastatic and angiogenic action of Ang-A. Combined treatment with Ang-A treated and U0126 exerted a synergistic inhibitory effect on the expression of MMP2 and VEGFA and the metastatic and angiogenic properties of human cervical cancer cells.

Significance

These findings are the first to indicate that in human cervical cancer cells, Ang-A exerts anti-metastatic and anti-angiogenic effects via targeting the miR-29a-3p/MMP2/VEGFA axis, mediated through the ERK pathway.

中文翻译：

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Angelol-A 通过调节靶向 MMP2/VEGFA 轴的磷酸化-ERK/miR-29a-3p 对人宫颈癌细胞发挥抗转移和抗血管生成作用

 目标

Angelol-A (Ang-A) 是一种香豆素类化合物，是从Angelica pubescens f. 的根中分离出来的。双锯齿。然而，AA对宫颈癌细胞的抗肿瘤作用及其分子机制尚不清楚。

 主要方法

使用MTT测定法测定细胞活力，并通过流式细胞术PI染色评估细胞周期阶段。评估使用/不使用 Antago-miR-29a-3p（miR-29a-3p 抑制剂）或 U0126（MEK 抑制剂）处理的 Ang-A 处理的细胞的 miR-29a-3p 表达、体外迁移/侵袭和血管生成分别使用 qRT-PCR、趋化性测定和管形成测定。使用适用的抗体通过蛋白质印迹分析测定丝裂原激活蛋白激酶/MMP2/MMP9/VEGFA的表达。

 主要发现

Ang-A 显着抑制人宫颈癌细胞中 MMP2 和 VEGFA 的表达、细胞迁移和侵袭运动。条件培养基抑制 HUVEC 中的管形成。 Ang-A 主要通过上调针对 VEGFA-3' UTR 的 miR-29a-3p 的表达来抑制侵袭性运动和血管生成。使用 Antago-miR-29a-3p 证实了 miR-29a-3p 的作用，它逆转了人宫颈癌细胞中 Ang-A 抑制的 MMP2 和 VEGFA 表达、侵袭性运动和血管生成。 ERK 通路参与介导 Ang-A 的转移和血管生成作用。 Ang-A处理和U0126联合治疗对MMP2和VEGFA的表达以及人宫颈癌细胞的转移和血管生成特性产生协同抑制作用。

 意义

这些发现首次表明，在人宫颈癌细胞中，Ang-A 通过靶向 miR-29a-3p/MMP2/VEGFA 轴（通过 ERK 通路介导）发挥抗转移和抗血管生成作用。