Berberine Suppresses EMT in Liver and Gastric Carcinoma Cells through Combination with TGFβR Regulating TGF‐β/Smad Pathway

Abstract

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.

Berberine Suppressed the Progression of Human Glioma Cells by Inhibiting the TGF-β1/SMAD2/3 Signaling Pathway

“… 20 Berberine also suppressed the EMT in hepatocellular carcinoma and gastric carcinoma cell lines by upregulating p-SMAD2 and downregulating p-SMAD3. 21 However, the relationship between berberine and the TGF-β/SMAD signaling pathway in glioma is still unclear.…”

The Core-Targeted RRM2 Gene of Berberine Hydrochloride Promotes Breast Cancer Cell Migration and Invasion via the Epithelial–Mesenchymal Transition

“…As a natural molecule, BBR has been demonstrated in the bit proliferation and metastasis of various cancers [ 7 , 8 , 9 , 10 , 27 ]. However, previous research on BBR in BRCA cells was limited to verifying known oncogenic pathways and targets, and the critical information for drug development was missing.…”

“…Moreover, BBR showed good antitumor activity. Several types of cancer cells are directly affected by it [ 7 , 8 , 9 , 10 ]. However, as with many natural therapeutic products, there is a lack of high-quality scientific evidence regarding the core pathways and targets of BBR.…”

High-content screening of active components of Traditional Chinese Medicine inhibiting TGF-β-induced cell EMT

“…Further experiments showed that CPT, DMC, ART, SNP and BER increased E-cadherin expression, decreased expression of Vimentin and α-SMA, enhanced cell adhesion and weakened cell migration. CPT, ART and BER have been previously reported to inhibit the EMT [ 33 , 34 , 35 ]. Identification of the inhibitory actions of DMC and SNP on the TGF-β1-induced EMT in A549 cells is a novel finding.…”