Berberine Suppresses EMT in Liver and Gastric Carcinoma Cells through Combination with TGFβR Regulating TGF-β/Smad Pathway.
Berberine Suppresses EMT in Liver and Gastric Carcinoma Cells through Combination with TGFβR Regulating TGF-β/Smad Pathway.
复制标题
小檗碱通过与调节 TGF-β/Smad 通路的 TGFβR 组合抑制肝癌和胃癌细胞中的 EMT。
DOI：
10.1155/2021/2337818
复制
发表时间：
2021
影响因子：
--
通讯作者：
Tan Y
中科院分区：
文献类型：
--
作者：
Du H;Gu J;Peng Q;Wang X;Liu L;Shu X;He Q;Tan Y
小檗碱（Berberine，BBR）是从黄连属植物中提取的一种天然生物碱，具有抗癌活性.有研究发现它能抑制黑色素瘤、神经母细胞瘤等肿瘤细胞的上皮间质转化（EMT）。然而，BBR是否能逆转肝细胞癌（HCC）和胃癌（GC）的EMT尚不清楚。在我们的研究中，BBR抑制HepG 2，MGC 803和SGC 7901细胞的迁移和侵袭，并呈剂量依赖性。转录测序分析表明，波形蛋白，MMP和Smad 3下调，但Smad 2，Smad 6，TAB 2，ZO-1和claudin 7上调时，与BBR处理。GO Enrichment分析KEGG信号通路显示，BBR在12 h显著抑制TGF-β/Smad信号通路，24 h抑制PI 3 K/Akt和Wnt/β-catenin信号通路，这两条信号通路与细胞增殖、迁移和EMT密切相关。通过Western Blot验证转录组测序分析的结果。结果表明，BBR处理后，上皮细胞标志物E-cadherin和ZO-1的表达显著增加，间充质细胞标志物N-cadherin和Vimentin的表达显著下降，转录因子Snail和Slug的表达显著下降。BBR的作用与PI 3 K抑制剂LY 294002和TGF-β受体抑制剂SB 431542的作用相似。BBR处理后，β-catenin和AKT、Smad 2和Smad 3的磷酸化水平均呈剂量依赖性变化，其中p-Smad 2上调，其余均下调。BBR分别与LY或SB联用可增强两种抑制剂的抑制作用。IGF-1和TGF-β分别是PI 3 K/AKT和TGF-β/Smad的激活剂，可逆转BBR的抗EMT作用。分子对接结果表明，BBR与TGF-β受体I（TGFβR1）具有较高的亲和力，结合能为-7.5 kcal/mol，优于TGFβR1的配体。虽然BBR与TGF-β受体II（TGFβR2）的亲和力低于TGFβR2的原始配体，但获得了更可观的负结合能（−8.54 kcal/mol）。BBR上调了p-Smad 2的表达，这与其他报道不同，表明Smad 2的功能相对复杂。BBR与SB联合应用可增强抑制剂对EMT的抑制作用，说明BBR与SB结合TGF β R的氨基酸位点不同，两者不存在竞争性结合。我们的实验证明BBR通过与TGFβR1和TGβ FR 2结合，抑制TGF-β/Smad，进而抑制PI 3 K/AKT等信号通路，增加p-Smad 2，降低p-Smad 3，从而抑制肿瘤细胞的EMT、转移和侵袭。BBR对三种肿瘤细胞的作用相似。
Berberine (BBR), a natural alkaloid derived from Coptis, has anticancer activity. Some researchers have found that it could restrain epithelial-mesenchymal transition (EMT) of melanoma, neuroblastoma, and other tumor cells. However, it is unclear whether BBR can reverse EMT in hepatocellular carcinoma (HCC) and gastric carcinoma (GC). In our study, BBR inhibited the migration and invasion of HepG2, MGC803, and SGC7901 cells in a dose-dependent manner. Transcription sequencing assays showed that Vimentin, MMP, and Smad3 were downregulated, but Smad2, Smad6, TAB2, ZO-1, and claudin 7 were upregulated when treated with BBR. GO Enrichment analysis of KEGG pathway showed that BBR significantly inhibited TGF-β/Smad at 12 h, then, PI3K/Akt and Wnt/β-catenin signaling pathways at 24 h, which were closely related to the proliferation, migration, and EMT. The results of the transcriptome sequencing analysis were verified by Western Blot. It showed that the expression of epithelial marker E-cadherin and ZO-1 remarkably augmented with BBR treatment, as well as declined mesenchymal markers, including N-cadherin and Vimentin, decreased transcription factor Snail and Slug. The effects of BBR were similar to those of the PI3K inhibitor LY294002 and TGF-β receptor inhibitor SB431542. Furthermore, β-catenin and phosphorylation of AKT, Smad2, and Smad3 were changed dose-dependently by BBR treatment, which upregulated p-Smad2 and downregulated the others. Combined with LY or SB, respectively, BBR could enhance the effects of the two inhibitors. Simultaneously, IGF-1 and TGF-β, which is the activator of PI3K/AKT and TGF-β/Smad, respectively, could reverse the anti-EMT effect of BBR. The Molecular Docking results showed BBR had a high affinity with the TGF-β receptor I (TGFβR1), and the binding energy was -7.5 kcal/mol, which is better than the original ligand of TGFβR1. Although the affinity of BBR with TGF-β receptor II (TGFβR2) was lower than the original ligand of TGFβR2, the more considerable negative binding energy (−8.54 kcal/mol) was obtained. BBR upregulated p-Smad2, which was different from other reports, indicating that the function of Smad2 was relatively complex. Combination BBR with SB could enhance the effect of the inhibitor on EMT, and the results indicated that BBR binding to TGFβR was not competitive with SB to TGFβR since different binding amino acid sites. Our experiments demonstrated BBR increased p-Smad2 and decreased p-Smad3 by binding to TGFβR1 and TGβFR2 inhibiting TGF-β/Smad, then, PI3K/AKT and other signaling pathways to restrain EMT, metastasis, and invasion in tumor cells. The effect of BBR was similar on the three tumor cells.
登录
查看更多内容
影响因子：
3.6
作者：
Chu, Shu-Chen;Yu, Cheng-Chia;Chen, Pei-Ni
通讯作者：
Chen, Pei-Ni
影响因子：
4.2
作者：
Huang, Chao;Liu, Hong;Wen, Bin
通讯作者：
Wen, Bin
影响因子：
5.6
作者：
Huang, Chao;Tao, Liang;Pang, Zuoliang
通讯作者：
Pang, Zuoliang
cPLA2α在肝癌细胞上皮间质转化过程中激活PI3K/AKT并抑制Smad2/3
影响因子：
9.7
作者：
Fu, Hui;He, Yuchao;Guo, Hua
通讯作者：
Guo, Hua
影响因子：
--
作者：
Kim, Sangmin;Lee, Jeongmin;Lee, Jeong Eon
通讯作者：
Lee, Jeong Eon