Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, School of Life Sciences, Jilin University, Changchun 130012, China.
Biomolecules. 2020 Mar 31;10(4):528. doi: 10.3390/biom10040528.
(1) Background: Epithelial-mesenchymal transition (EMT) is an essential step for cancer metastasis; targeting EMT is an important path for cancer treatment and drug development. NF-κB, an important transcription factor, has been shown to be responsible for cancer metastasis by enhancing the EMT process. Our previous studies showed that (20S)Ginsenoside Rh2 (G-Rh2) inhibits NF-κB activity by targeting Anxa2, but it is still not known whether this targeted inhibition of NF-κB can inhibit the EMT process. (2) Methods: In vivo (20S)G-Rh2-Anxa2 interaction was assessed by cellular thermal shift assay. Protein interaction was determined by immuno-precipitation analysis. NF-κB activity was determined by dual luciferase reporter assay. Gene expression was determined by RT-PCR and immuno-blot. EMT was evaluated by wound healing and Transwell assay and EMT regulating gene expression. (3) Results: Anxa2 interacted with the NF-κB p50 subunit, promoted NF-κB activation, then accelerated mesenchymal-like gene expression and enhanced cell motility; all these cellular processes were inhibited by (20S)G-Rh2. In contrast, these (20S)G-Rh2 effect were completely eliminated by overexpression of Anxa2-K301A, an (20S)G-Rh2-binding-deficient mutant of Anxa2. (4) Conclusion: (20S)G-Rh2 inhibited NF-κB activation and related EMT by targeting Anxa2 in MDA-MB-231 cells.
(1) 背景：上皮-间质转化（EMT）是癌症转移的重要步骤；靶向 EMT 是癌症治疗和药物开发的重要途径。NF-κB 是一种重要的转录因子，通过增强 EMT 过程被证明与癌症转移有关。我们之前的研究表明，（20S）人参皂苷 Rh2（G-Rh2）通过靶向 Anxa2 抑制 NF-κB 活性，但尚不清楚这种针对 NF-κB 的靶向抑制是否能抑制 EMT 过程。
(2) 方法：通过细胞热转移分析评估体内（20S）G-Rh2-Anxa2 相互作用。通过免疫沉淀分析确定蛋白相互作用。通过双荧光素酶报告基因检测法测定 NF-κB 活性。通过 RT-PCR 和免疫印迹测定基因表达。通过划痕愈合和 Transwell 测定以及 EMT 调节基因表达评估 EMT。
(3) 结果：Anxa2 与 NF-κB p50 亚基相互作用，促进 NF-κB 激活，然后加速间充质样基因表达并增强细胞迁移；所有这些细胞过程都被（20S）G-Rh2 抑制。相反，通过过表达 Anxa2-K301A（Anxa2 的一种与（20S）G-Rh2 结合缺陷的突变体），完全消除了（20S）G-Rh2 的这些作用。
(4) 结论：（20S）G-Rh2 通过在 MDA-MB-231 细胞中靶向 Anxa2 抑制 NF-κB 激活及其相关的 EMT。
