Dataset Information

A83-01 inhibits TGF-?-induced upregulation of Wnt3 and epithelial to mesenchymal transition in HER2-overexpressing breast cancer cells.

ABSTRACT: The aim of this study is to investigate the mechanisms of interactions between TGF-? and Wnt/?-catenin pathways that induce and regulate EMT and promote breast cancer cells to become resistant to treatment.The effect of TGF-? on Wnt/?-catenin signaling pathway was examined by using a human Wnt/?-catenin-regulated cDNA plate array and western blot analysis. The interaction of Twist at promoter of Wnt3 was examined by chromatin immunoprecipitation (ChIP) assay. Secreted Wnt3 level was determined by ELISA assay.HER2-overexpressing breast cancer cells treated with TGF-? have a reduced response to trastuzumab and exhibited EMT-like phenotype. The TGF-?-induced EMT in HER2-cells was concordant with upregulation of Wnt3 and ?-catenin pathways. The TGF-?-induced induction of Wnt3 during EMT was found to be Smad3-dependent. ChIP analysis identified occupancy of Twist at promoter region of Wnt3. Knock-down of Twist by shRNA confirmed the significance of Twist in response to TGF-? regulating Wnt3 during EMT. Subsequently, TGF-?-induced matrix metalloproteinases, MMP1, MMP7, MMP9, MMP26, Vascular endothelial growth factors (VEGF), and activation of Wnt/?-catenin signaling were repressed by the shRNA treatment. TGF-?R1 ALK5 kinase inhibitor, A83-01 can effectively prevent the TGF-?-induced Twist and Wnt3. Co-treating A83-01 and trastuzumab inhibited TGF-?-induced cell invasion significantly in both trastuzumab responsive and resistant cells.Our data demonstrated an important interdependence between TGF-? and Wnt/?-catenin pathways inducing EMT in HER2-overexpressing breast cancer cells. Twist served as a linkage between the two pathways during TGF-?-induced EMT. A83-01 could inhibit the TGF-?-initiated pathway interactions and enhance HER2-cells response to trastuzumab treatment.

SUBMITTER: Wu Y

PROVIDER: S-EPMC5427117 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

A83-01 inhibits TGF-β-induced upregulation of Wnt3 and epithelial to mesenchymal transition in HER2-overexpressing breast cancer cells.

Wu Yanyuan Y Tran Trinh T Dwabe Sami S Sarkissyan Marianna M Kim Juri J Nava Miguel M Clayton Sheilah S Pietras Richard R Farias-Eisner Robin R Vadgama Jaydutt V JV

Breast cancer research and treatment 20170323 3

<h4>Purpose</h4>The aim of this study is to investigate the mechanisms of interactions between TGF-β and Wnt/β-catenin pathways that induce and regulate EMT and promote breast cancer cells to become resistant to treatment.<h4>Methods</h4>The effect of TGF-β on Wnt/β-catenin signaling pathway was examined by using a human Wnt/β-catenin-regulated cDNA plate array and western blot analysis. The interaction of Twist at promoter of Wnt3 was examined by chromatin immunoprecipitation (ChIP) assay. Secr ...[more]

PMID: 28337662

Similar Datasets

Project description:Inflammatory bowel disease (IBD), which consists of Crohn's disease (CD) and ulcerative colitis (UC), is a chronic, inflammatory disorder of the gastro-intestinal tract with unknown etiology. Current evidence suggests that intestinal epithelial cells (IECs) is prominently linked to the pathogenesis of IBD. Therefore, maintaining the intact of epithelium has potential roles in improving pathophysiology and clinical outcomes of IBD. MicroRNAs (miRNAs) act as post-transcriptional gene regulators and regulate many biological processes, including embryonal development, cell differentiation, apoptosis and proliferation. In this study, we found that miR-200b decreased significantly in inflamed mucosa of IBD, especially for UC, when compared with their adjacent normal tissue. Simultaneously, we also found that the genes of E-cadherin and cyclin D1 were reduced significantly and correlated positively to the miR-200b. In addition, the upregulation of transforming growth factor-beta 1 (TGF-?1) was inversely correlated to the miR-200b in IBD. To investigate the possible roles of miR-200b in IECs maintaining, we used TGF-?1 to induce epithelial-mesenchymal transition (EMT) in IEC-6 initially. After sustained over-expressing miR-200b in IEC-6, the EMT was inhibited significantly that was characterized by downregulation of vimentin and upregulation of E-cadherin. Furthermore, we found that miR-200b enhanced E-cadherin expression through targeting of ZEB1, which encode transcriptional repressors of E-cadherin. SMAD2 was found to act as a target of miR-200b with direct evidence that miR-200b binding to the 3' UTR of SAMD2 and the ability of miR-200b to repress SMAD2 protein expression. With SMAD2 depletion, the expression of vimentin decreased correspondingly, which suggested miR-200b might reduce vimentin through regulating the SMAD2. With endogenous over-expression of miR-200b, the proliferation of IEC-6 cells increased significantly by increasing S-phase entry and promoting expression of the protein cyclin D1. Summarily, our study suggested a potential role for mir-200b in maintaining intact of intestinal epithelium through inhibiting EMT and promoting proliferation of IECs.

Dataset Information

A83-01 inhibits TGF-?-induced upregulation of Wnt3 and epithelial to mesenchymal transition in HER2-overexpressing breast cancer cells.

Publications

A83-01 inhibits TGF-β-induced upregulation of Wnt3 and epithelial to mesenchymal transition in HER2-overexpressing breast cancer cells.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets