MMP-2 and MMP-9 mediate cigarette smoke extract-induced epithelial-mesenchymal transition in airway epithelial cells via EGFR/Akt/GSK3β/β-catenin pathway: Amelioration by fisetin
MMP-2 and MMP-9 mediate cigarette smoke extract-induced epithelial-mesenchymal transition in airway epithelial cells via EGFR/Akt/GSK3β/β-catenin pathway: Amelioration by fisetin
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DOI：
10.1016/j.cbi.2019.108846
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发表时间：
2019-12-01
影响因子：
5.1
通讯作者：
Yadav, Umesh C. S.
中科院分区：
文献类型：
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作者：
Agraval, Hina;Yadav, Umesh C. S.
基质金属蛋白酶(MMPs)已被认为与EMT有关，但其在香烟烟雾诱导的呼吸道上皮细胞EMT中的调控作用尚不清楚。因此，我们利用A549肺上皮细胞和人小气道上皮细胞(SAEC)研究了MMP2和-9在香烟烟雾提取物(CSE)诱导的EMT中的潜在作用。用不同浓度的CSE处理细胞，用四甲基偶氮唑蓝、台盼蓝比色法、吖啶橙-溴化乙锭比色法、明胶酶谱、Western blotting、免疫荧光、Boyden小室实验、创面愈合实验和气液界面培养等方法检测EMT后细胞和分子的变化。结果表明，CSE在增加细胞毒活性的同时，也增加了基质金属蛋白酶-2和-9的表达和活性。CSE进一步改变了EMT标志物如E-钙粘蛋白、N-钙粘蛋白、波形蛋白，以及EMT的分子调节物如β-连环蛋白和pGSK-3β。此外，CSE还上调了呼吸道上皮细胞中EGFR、AKT和ERK1/2的表达。已知的基质金属蛋白酶-2和-9的抑制剂SB-3CT改变和逆转了EMT和激酶的标志物的表达，证实了基质金属蛋白酶-2和-9在CSE诱导的EMT中的作用。Fisetin是一种植物来源的生物黄酮类化合物，它也以与SB-3CT类似的方式逆转了EMT标记和分子调节因子的表达。综上所述，本研究强调了MMP-2和-9在CSE诱导的EMT中的作用，并通过EGFR/AKT/ERK/β-catenin轴调控其分子级联反应，该信号通路可被基质金属蛋白酶-2和-9抑制剂和非赛汀恢复。非瑟素对CSE诱导的呼吸道上皮细胞MMPs活性的调节作用尚不清楚，我们的研究提示其在CSE诱导的肺上皮细胞EMT中具有潜在的治疗作用。
Matrix metalloproteinases (MMPs) have been implicated in EMT but their role in the regulation of cigarette smoke-induced EMT in airway epithelium is not clear. We have therefore investigated the potential role of MMP2 and -9 in cigarette smoke extract (CSE) induced EMT using A549 lung epithelial cells and human small airway epithelial cells (SAEC). The cells were treated with different concentration of CSE, and MTT and trypan blue assays, acridine orange-ethidium bromide assay, gelatin zymography, Western blotting, immunofluorescence studies, Boyden-chamber assay, wound healing assay and air-liquid interface (ALI) culture were used to assess different cellular and molecular changes associated with EMT. The results depict that CSE increased the cytotoxicity along with a concurrent increase in the expression and activity of MMP-2 and -9. CSE further altered EMT markers like E-cadherin, N-cadherin, vimentin, and the molecular modulators of EMT such as beta-catenin and pGSK-3 beta. Further, CSE also upregulated EGFR, AKT, and ERK1/2 in airway epithelial cells. SB-3CT, a known inhibitor of MMP-2 and -9, altered and reversed the expression of markers of EMT and kinases, validating the role of MMP-2 and -9 in CSE-induced EMT. Fisetin, a plant-derived bioflavonoid, also reversed the expression of EMT markers and molecular regulators in a similar fashion as SB-3CT. In summary, this study highlights the role of MMP-2 and -9 in CSE-induced EMT and curate its molecular cascade through EGFR/AKT/ERK/beta-catenin axis, which could be restored by MMP-2 and -9 inhibitor and fisetin. Fisetin is hitherto unknown to modulate CSE-induced MMPs activity in airway epithelial cells, and our study suggests its potential role as a therapeutic approach in CSE-induced EMT in lung epithelial cells.