Osthole inhibited TGF β-induced epithelial-mesenchymal transition (EMT) by suppressing NF-κB mediated Snail activation in lung cancer A549 cells
Osthole inhibited TGF β-induced epithelial-mesenchymal transition (EMT) by suppressing NF-κB mediated Snail activation in lung cancer A549 cells
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DOI：
10.1080/19336918.2016.1259058
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发表时间：
2017-01-01
影响因子：
3.2
通讯作者：
Chen, Xiuping
中科院分区：
文献类型：
--
作者：
Feng, Haitao;Lu, Jin-Jian;Chen, Xiuping
上皮间质转化（EMT），即上皮细胞向间质细胞的转分化，与转移有关，并为癌症治疗提供了新的策略。蛇床子素 (OST) 是中药蛇床子的主要活性成分，据报道可抑制癌症转移，但其机制尚不清楚。在这里，我们研究了OST对TGF-β1诱导的A549细胞EMT的抑制作用和机制。在不存在和存在 OST 的情况下用 TGF-β1 处理细胞。用显微镜观察形态变化。通过Western blotting和实时PCR测定蛋白质和mRNA的表达。通过免疫荧光检测蛋白质定位。通过基质胶、伤口愈合和Transwell实验测定粘附、迁移和侵袭。 TGF-β 1 处理诱导细胞发生纺锤形改变，N-钙粘蛋白、波形蛋白、NF-κ B p65 上调，以及 E-钙粘蛋白下调。 TGF-β1处理后观察到E-钙粘蛋白和N-钙粘蛋白的膜表达和mRNA表达失调。 TGF-β 1 增加迁移和侵袭的能力并触发 NF-κ B p65 的核转位。这些改变被 OST 显着抑制。此外，PDTC（一种 NF-κ B 抑制剂）也表现出类似的效果。此外，OST 显着抑制 TGF-β 1 诱导的 Snail 表达，沉默 Snail 部分逆转 TGF-β 1 诱导的 EMT 生物标志物，而不影响 NF-kappa B p-65。总之，OST 通过灭活 A549 细胞中的 NF-κ B-Snail 通路来抑制 TGF-β 1 诱导的 EMT、粘附、迁移和侵袭。这项研究为OST的抗转移作用提供了新的分子机制。
Epithelial-mesenchymal transition (EMT), the transdifferentiation of epithelial cells into mesenchymal cells, has been implicated in the metastasis and provides novel strategies for cancer therapy. Osthole (OST), a dominant active constituent of Chinese herb Cnidium monnieri, has been reported to inhibit cancer metastasis while the mechanisms remains unclear. Here, we studied the inhibitory effect and mechanisms of OST on TGF-beta 1-induced EMT in A549 cells. Cells were treated with TGF-beta 1 in the absence and presence of OST. The morphological alterations were observed with a microscopy. The protein and mRNA expressions were determined by Western blotting and real-time PCR. The protein localization was detected with immunofluorescence. The adhesion, migration, and invasion were determined by Matrigel, wound-healing, and Transwell assays. TGF-beta 1 treatment induced spindle-shaped alterations of cells, upregulation of N-cadherin, Vimentin, NF-kappa B p65, and downregulation of E-cadherin. Dysregulated membrane expression and mRNA expression of E-cadherin and N-cadherin were observed after TGF-beta 1 treatment. TGF-beta 1 increased abilities of migration and invasion and triggered the nuclear translocation of NF-kappa B p65. These alterations were dramatically inhibited by OST. Furthermore, PDTC, a NF-kappa B inhibitor, showed similar effects. In addition, TGF-beta 1-induced expression of Snail was significantly inhibited by OST and silenced Snail partially reversed TGF-beta 1-induced EMT biomarkers without affecting NF-kappa B p-65. In conclusion, OST inhibited TGF-beta 1-induced EMT, adhesion, migration, and invasion through inactivation of NF-kappa B-Snail pathways in A549 cells. This study provides novel molecular mechanisms for the anti-metastatic effect of OST.