Time course studies on the appearance of TGF-beta indicated that most of the active TGF-beta was generated within the first 12 h after the establishment of co-cultures. The generation of TGF-beta in co-cultures stimulated the production of the protease inhibitor plasminogen activator inhibitor-1 (PAI-1).
The target sequences were proven to be highly specific and effective for suppressing both TGF-β1 mRNA and protein expression in cells after infection with an adenovirus expressing TGF-β1 short hairpin RNA (shRNA). A single base pair change in the shRNA sequence completely abrogated the suppressive effect on TGF-β1.
Nat Commun PMC5945639 Nat Commun.2018; 9: 1845. Published online 2018 May 10. doi: 10.1038/s41467-018-03962-x PMCID: PMC5945639 PMID: 29748571 TGF-β induces miR-100 and miR-125b but blocks let-7a through LIN28B controlling PDAC progression
Total RNA from cultured cells was extracted using TRI Reagent (Sigma) following the manufacturer’s instructions including DNase I treatment. qRT-PCR of mature miRNAs was performed using TaqMan Small RNA Assays (Applied Biosystems) with assays for hsa-miR-100 (000437), hsa-miR-125b (000449), hsa-let-7a (000377), has-miR-200a (000502).
TGF-beta inhibits proliferation and induces apoptosis in various cell types, and accumulation of loss-of-function mutations in the TGF-beta receptor or Smad genes classify the pathway as a tumor suppressor in humans.
Transforming growth factor beta (TGF-β) is a multipotent growth factor affecting cell differentiation, proliferation, apoptosis and matrix production.
TGF-β induces the demethylation of H3K27me3 in Snail1 promoter and overexpresses Snail1, which cause EMT. Referring to these results, it's possible that the mechanism of TGF-β to induce EMT is correlated with Smad, which is the upstream molecule of Snail in the TGF-β signaling cascade21,22.
TGFb in the immune system TGFb1 is conventionally regarded as an anti-inflammatory agent, not least because of the severe immune pathology seen in TGFb1 knockout mice or in mice with impaired TGFb signalling in T cells [6–8].
Transforming growth factor β (TGF-β) is a highly pleiotropic cytokine that plays an important role in wound healing, angiogenesis, immunoregulation and cancer. The cells of the immune system produce the TGF-β1 isoform, which exerts powerful anti-inflammatory functions, and is a master regulator of the immune response.
TGF-β is secreted by many cell types, including macrophages, in a latent form in which it is complexed with two other polypeptides, latent TGF-beta binding protein (LTBP) and latency-associated peptide (LAP). Serum proteinases such as plasmin catalyze the release of active TGF-β from the complex.
Addition of TGF beta 1 produced a dramatic reorganization of apical F-actin and development of stress fibers, as well as the loss of normal cell border-associated ZO-1 distribution. The effects of TGF beta 1 were blocked by the neutralizing antibodies to TGF beta 1.
Results: TGF-beta treatment was associated with morphologic and phenotypic changes typical of epithelial-mesenchymal transition (EMT) including increased fibrogenesis in all renal cell types and decreased E-cadherin expression in tubular cells.
EMT has been shown to be induced by androgen deprivation therapy in metastatic prostate cancer. Activation of EMT programs via inhibition of the androgen axis provides a mechanism by which tumor cells can adapt to promote disease recurrence and progression.
What does it mean if your TGF-b1 result is too high? - TGF B-1 is often chronically over-expressed in disease states, including cancer, fibrosis and inflammation. - TGF B-1 is moderately to extremely high in Chronic Inflammatory Response Syndrome due to water-damaged buildings (CIRS).
One of a group of related proteins made by leukocytes (white blood cells) and other cells in the body. Interleukins regulate immune responses. Interleukins made in the laboratory are used as biological response modifiers to boost the immune system in cancer therapy.
TGF-β also plays a major role under inflammatory conditions. TGF-β in the presence of IL-6 drives the differentiation of T helper 17 (Th17) cells, which can promote further inflammation and augment autoimmune conditions [15].
Among the soluble factors that regulate skeletal muscle function, Transforming Growth Factor type Beta 1 (TGF-b1) is one of the most studied. This factor inhibits myogenesis and regeneration by regulating the activity and function of satellite cells (SCs). Indeed, TGF-b has a central role in muscle pathologies in which there is development of fibrosis and/or atrophy of skeletal muscle. Thus, in this review we present the critical and recent antecedents regarding the mechanisms and cellular targets involved in the effects of TGF-b1 in the muscle, in pathological processes such as the inhibition of regeneration, fibrosis and atrophy. In addition, an update on the development of new strategies with therapeutic potential to inhibit the deleterious actions of TGF-b in skeletal muscle is discussed.
... Transforming growth factor-β1 (TGF-β1) is a potent fibrogenic cytokine in many tissues and organs, including the lungs, kidneys, liver, heart, and skin [10] [11] [12] [13]. To date, it has been widely accepted that TGF-β1 is also associated with fibrosis in skeletal muscle [14] [15] [16]. Elevated TGF-β1 levels are observed in dystrophic skeletal muscle and shortly after skeletal muscle injury [14] [15] [16] [17]. Research has shown that elevated expression of TGF-β1 accounts for the initiation and maintenance of fibrosis in muscular dystrophies [14,15]. ...
... Fibrosis is defined by the enhanced deposition of extracellular matrix proteins, mainly collagens, in the basement membrane and interstitial tissue. Additionally, it has been reported that fibrogenesis is mainly mediated by TGF-β signaling pathway (Chen et al., 2005; Salvadori et al., 2005), which promotes fibrosis via activation of the small body size (SMA)-and mothers against decapentaplegic (MAD)-related protein signaling pathway ( Gosselin et al., 2004; Decologne et al., 2007 ) to induce the expression of fibrogenic genes (Kennedy et al., 2008) such as procollagen as well as the enzymes that catalyze the collagen crosslinking (Massague and Chen, 2000). In this study there was a greater expression of TGF-β in ON compared to a CTL fetal muscle, which is consistent with a greater expression of collagen III also observed in ON compared to CTL fetal muscle. ...
Sulforaphane (SFN), an activator of NF-E2-related factor 2 (Nrf2), has been found to have anti-fibrotic effect on liver and lung. However, its effects on dystrophic muscle fibrosis remain unknown. This work was undertaken to evaluate the effects of SFN-mediated activation of Nrf2 on dystrophic muscle fibrosis . 3-month-old male mdx mice were treated with SFN by gavage (2 mg/kg body weight per day) for 3 months. The experimental results demonstrated that SFN remarkably attenuated skeletal and cardiac muscle fibrosis as indicated by reduced Sirius red staining and immuno-staining of ECM. Moreover, SFN significantly inhibited TGF-β/Smad signaling pathway and ameliorated pro-fibrogenic gene and protein expression, such as α-SMA, fibronectin, collagen I, PAI-1 and TIMP-1 in Nrf2 dependent manner. Further, SFN significantly decreased the expression of inflammatory-cytokines CD45, TNF-α and IL-6 in mdx mice. In conclusion, these results show that SFN can attenuate dystrophic muscle fibrosis by Nrf2-mediated inhibition of TGF-β/Smad signaling pathway, which indicate Nrf2 may represent a new target for dystrophic muscle fibrosis.
The mdx mouse is a good genetic and molecular murine model for Duchenne Muscular Dystrophy (DMD), a progressive and devastating muscle disease. However, this model is inappropriate for testing new therapies due to its mild phenotype. Here, we transferred the mdx mutation to the 129/Sv strain with the aim to create a more severe model for DMD. Unexpectedly, functional analysis of the first three generations of mdx129 showed a progressive amelioration of the phenotype, associated to less connective tissue replacement, and more regeneration than the original mdxC57BL. Transcriptome comparative analysis was performed to identify what is protecting this new model from the dystrophic characteristics. The mdxC57BL presents three times more differentially expressed genes (DEGs) than the mdx129 (371 and 137 DEGs respectively). However, both models present more overexpressed genes than underexpressed, indicating that the dystrophic and regenerative alterations are associated with the activation rather than repression of genes. As to functional categories, the DEGs of both mdx models showed a predominance of immune system genes. Excluding this category, the mdx129 model showed a decreased participation of the endo/exocytic pathway and homeostasis categories, and an increased participation of the extracellular matrix and enzymatic activity categories. Spp1 gene overexpression was the most significant DEG exclusively expressed in the mdx129 strain. This was confirmed through relative mRNA analysis and osteopontin protein quantification. The amount of the 66 kDa band of the protein, representing the post-translational product of the gene, was about 4,8 times higher on western blotting. Spp1 is a known DMD prognostic biomarker, and our data indicate that its upregulation can benefit phenotype. Modeling the expression of the DEGs involved in the mdx mutation with a benign course should be tested as a possible therapeutic target for the dystrophic process.
Transforming growth factor (TGF)-β regulates the expression of matrix metalloproteinases (MMPs) and components of the extracellular matrix, thereby profoundly affecting the microenvironment of cells including cancerous ones.
Since its discovery as a cytokine secreted from transformed fibroblasts with the ability to transform normal fibroblasts in vitro, transforming growth factor (TGF)-β has been intensively studied as a cytokine closely related to tumourigenesis.
Transforming growth factor-β signalling targets expression of MMP-10. As shown in Figures 1a and b, MMP-10 was induced by TGF-β at mRNA and protein levels in both mouse and human mammary epithelial cell lines, NMuMG and MCF10A.
Expression of most MMPs is normally low in tissues and is induced where and when the extracellular matrix is actively remodelled.
Mammary epithelial cell lines were obtained from the American Type Cell Collection. NMuMG mouse mammary epithelial cells were maintained as described previously ( Kanome et al., 2007 ).
We thank Dr Eric N Olson (University of Texas Southwestern Medical School, Dallas, TX, USA) for his generous gift of the MMP-10 promoter luciferase constructs. We also thank Ms E Kaneko and Ms T Kaneko for contributing to this work as a project for their bachelor's degree.
Supplementary Information accompanies the paper on the Oncogene website ( http://www.nature.com/onc)
The competitive endogenous RNA (ceRNA) hypothesis suggests an intrinsic mechanism to regulate biological processes. However, whether the dynamic changes of ceRNAs can modulate miRNA activities remains controversial. Here, we examine the dynamics of ceRNAs during TGF-β-induced epithelial-to-mesenchymal transition (EMT).
MicroRNAs (miRNAs) are ubiquitous post-transcriptional regulators that impact RNA stability and the rate of translation by pairing to complementary sites (referred to as miRNA response elements [MREs]) within target RNAs 1, 2, 3.
In a previous study, we profiled the transcriptional dynamics of TGF-β-induced EMT in A549 cells 22.
Here, we showed that a single mRNA dynamically induced during EMT could represent the vast majority of induced MREs and regulate EMT through a ceRNA effect.
A549 cells and MCF10A cells were obtained from cell bank of Chinese Academy of Sciences and have been test to be free of mycoplasma contamination. The identities of cells have been confirmed by STR profiling.
The miRNA-seq data reported in this study have been deposited in GEO under accession number GSE87358. All other data are available from the authors upon reasonable request.
The ODEs describing the EMT model with FN1 are provided in Supplemental Note.