Fisetin Suppresses the Proliferation and Metastasis of Renal Cell Carcinoma through Upregulation of MEK/ERK-Targeting CTSS and ADAM9
Fisetin Suppresses the Proliferation and Metastasis of Renal Cell Carcinoma through Upregulation of MEK/ERK-Targeting CTSS and ADAM9
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DOI：
10.3390/cells8090948
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发表时间：
2019-09-01
期刊：
影响因子：
6
通讯作者：
Chang, Horng-Rong
中科院分区：
文献类型：
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作者：
Hsieh, Min-Hong;Tsai, Jen-Pi;Chang, Horng-Rong
非瑟酮是一种天然黄酮类化合物，已知对多种癌症具有抗癌作用，但其在介导肾细胞癌(RCC)进展中的作用尚未阐明。用3-(4，5-西甲基噻唑-2-基)-2，5-二苯基四氮唑溴化法和碘化丙啶染色流式细胞仪检测细胞存活率、细胞毒性和细胞周期分布。体外迁移和侵袭实验检测细胞在体内的迁移和侵袭。用细胞迁移/侵袭相关蛋白进行人蛋白水解酶抗体芯片分析。Western blotting和定量逆转录聚合酶链式反应检测与细胞周期、细胞侵袭和丝裂原活化蛋白激酶(MAPK)信号通路相关的蛋白表达。我们发现，除了下调细胞周期蛋白d1和上调p21/p27外，非瑟素还通过将细胞周期停滞在G2/M期而显著抑制细胞活力。非瑟素通过下调CTSS和去整合素及金属蛋白酶9(ADAM9)抑制人肾癌细胞的迁移和侵袭。非瑟素还上调了786-O和Caki-1细胞中ERK的磷酸化水平。此外，MEK抑制剂(UO126)可通过ERK/CTSS/ADAM9通路降低非瑟素对肾癌细胞转移的抑制作用。非瑟素通过MEK/ERK信号通路下调CTSS和ADAM9，从而抑制肾癌细胞的增殖和转移。这些结果表明，非瑟素是一种很有前途的抗肾癌药物。
Fisetin, a natural flavonoid, is known to have anticarcinogenic effects against several cancers, but its role in mediating renal cell carcinoma (RCC) progression has not been delineated. Cell viability, cytotoxicity, and cell cycle distribution were measured using the 3-(4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and propidium iodide staining with flow cytometry. The in vitro migration and invasion assay was used to examine in vivo cell migration and invasion. Human protease antibody array analysis was conducted with cell migration/invasion-related proteins. Western blotting and quantitative reverse transcription polymerase chain reaction were used for assessing protein expression related to the cell cycle, cell invasion, and mitogen-activated protein kinase (MAPK) signaling pathway. We found that fisetin significantly inhibited cell viability through cell cycle arrest in the G2/M phase, in addition to downregulating cyclin D1 and upregulating p21/p27. Fisetin inhibited the migration and invasion of human RCC cells through the downregulation of CTSS and a disintegrin and metalloproteinase 9 (ADAM9). Fisetin also upregulated ERK phosphorylation in 786-O and Caki-1 cells. Furthermore, treatment with a MEK inhibitor (UO126) reduced the inhibitory effects of fisetin on the metastasis of RCC cells through the ERK/CTSS/ADAM9 pathway. Fisetin inhibits proliferation and metastasis of RCC cells by downregulating CTSS and ADAM9 through the MEK/ERK signaling pathway. These findings indicate that fisetin is a promising antitumor agent against RCC.