Inhibition of TGF-β2-induced migration and epithelial-mesenchymal transition in ARPE-19 by sulforaphane
Inhibition of TGF-β2-induced migration and epithelial-mesenchymal transition in ARPE-19 by sulforaphane
复制标题
DOI：
10.18240/ijo.2021.07.03
复制
发表时间：
2021-07-18
影响因子：
1.4
通讯作者：
Liu, Yang
中科院分区：
文献类型：
--
作者：
Huang, Yan-Bing;Liu, Ping-Ping;Liu, Yang
目的：目的：探讨莱菔硫烷（SFN）对转化生长因子β 2（TGF-β 2）刺激的ARPE-19细胞迁移和上皮-间质转化（EMT）的影响。通过进行乳酸脱氢酶测定（LDH）和3-（4，5-二甲基噻唑-2-基）-5-（3-羧基甲氧基苯基）-2-（4-磺基苯基）-2H-四唑（MTS）测定来评估SFN毒性，并且通过Transwell迁移测定来评估细胞迁移。使用免疫荧光分析测定ARPE-19细胞中肌动蛋白应力纤维的形成。免疫印迹分析法测定纤维连接蛋白和α-平滑肌肌动蛋白的表达沿着Smad和Akt磷酸化的程度。此外，SFN减弱了TGF-β 2诱导的肌动蛋白应力纤维的出现以及这些细胞中的纤连蛋白和α-平滑肌肌动蛋白表达。SFN还阻碍TGF-β 2刺激的Smad 2、Smad 3和Akt的磷酸化。结论：SFN抑制TGF-β 2诱导的ARPE-19细胞迁移和EMT，其机制可能与抑制肌动蛋白应力纤维的形成和Akt、Smad 2/3信号通路有关。
AIM: To investigate the effects of sulforaphane (SFN) on transforming growth factor (TGF)-beta 2 stimulated migration and epithelial-mesenchymal transition (EMT) in ARPE-19 cells.METHODS: ARPE-19 cells were cultured in the presence or absence of SFN or TGF-beta 2. SFN toxicity was assessed by performing a lactate dehydrogenase assay (LDH) and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4- sulfophenyl)-2H-tetrazolium (MTS) assays, and cell migration was evaluated by Transwell migration assay. Actin stress fiber formation in ARPE-19 cells was determined using immunofluorescence analysis. Immunoblotting analysis was used to determine fibronectin and alpha-smooth muscle actin expressions along with the degree of Smad and Akt phosphorylation.RESULTS: SFN inhibited ARPE-19 migration. Additionally, SFN attenuated TGF-beta 2-induced appearance of actin stress fibers as well as fibronectin and alpha-smooth muscle actin expressions in these cells. SFN also hindered the TGF-beta 2-stimulated phosphorylation of Smad2, Smad3, and Akt. SFN showed no cytotoxicity towards ARPE-19 cells.CONCLUSION: SFN inhibits TGF-beta 2-stimulated migration and EMT in ARPE-19 cells, probably by preventing the establishment of actin stress fibers and Akt and Smad2/3 signaling.