Xu Mengzhen, Cui Qinghua, Su Wen, Zhang Dan, Pan Jiaxu, Liu Xiangqi, Pang Zheng, Zhu Qingjun
College of Pharmaceutical Science, Shandong University of Traditional Chinese Medicine, Jinan 250355, China.
Innovative Institute of Chinese Medicine and Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan 250355, China.
Heliyon. 2022 Aug 15;8(8):e10238. doi: 10.1016/j.heliyon.2022.e10238. eCollection 2022 Aug.
The epithelial mesenchymal transition (EMT) has roles in metastasis and invasion during fibrotic diseases and cancer progression. Some Traditional Chinese Medicines (TCMs) have shown inhibitory effects with respect to the EMT. The current study attempted to establish a multiparametric high-content method to screen for active monomeric compounds in TCM with the ability to target cellular EMT by assessing phenotypic changes. A total of 306 monomeric compounds from the MedChemExpress (MCE) compound library were screened by the high-content screening (HCS) system and 5 compounds with anti-EMT activity, including camptothecin (CPT), dimethyl curcumin (DMC), artesunate (ART), sinapine (SNP) and berberine (BER) were identified. To confirm anti-EMT activity, expression of EMT markers was assessed by qRT-PCR and Western blotting, and cell adhesion and migration measured by cell function assays. The results revealed that CPT, DMC, ART, SNP and BER inhibited transforming growth factor-β1 (TGF-β1)-induced expression of vimentin and α-SMA, upregulated expression of E-cadherin, increased cell adhesion and reduced cell migration. In summary, by quantifying the cell morphological changes during TGF-β1-induced EMT through multi-parametric analysis, TCM compounds with anti-EMT activity were successfully screened using the HCS system, a faster and more economical approach than conventional methods.
上皮-间质转化（EMT）在纤维化疾病和癌症进展过程中的转移和侵袭中发挥作用。一些中药已显示出对EMT的抑制作用。本研究试图建立一种多参数高内涵方法，通过评估表型变化来筛选具有靶向细胞EMT能力的中药活性单体化合物。利用高内涵筛选（HCS）系统从MedChemExpress（MCE）化合物库中筛选了306种单体化合物，鉴定出5种具有抗EMT活性的化合物，包括喜树碱（CPT）、二甲基姜黄素（DMC）、青蒿琥酯（ART）、芥子碱（SNP）和小檗碱（BER）。为了确认抗EMT活性，通过qRT-PCR和蛋白质印迹法评估EMT标志物的表达，并通过细胞功能试验测量细胞黏附和迁移。结果显示，CPT、DMC、ART、SNP和BER抑制了转化生长因子-β1（TGF-β1）诱导的波形蛋白和α-平滑肌肌动蛋白（α-SMA）的表达，上调了E-钙黏蛋白的表达，增加了细胞黏附并减少了细胞迁移。总之，通过多参数分析定量TGF-β1诱导的EMT过程中的细胞形态变化，利用HCS系统成功筛选出具有抗EMT活性的中药化合物，这是一种比传统方法更快、更经济的方法。
