Transforming Growth Factor-beta Type Research Articles

Identification of Phytochemical Inhibitors Targeting VEGFA and TGFBR2 of Cervical Cancer: Insights from Virtual Screening, ADMET and DFT Studies

AbstractThe vascular endothelial growth factor A (VEGFA) and Transforming Growth Factor Beta type II receptor (TGFBR2) serves as a promising candidate in the treatment of cervical cancer, playing a crucial role in angiogenesis and tumorigenesis. Its involvement in cervical cancer has been well documented. The pursuit of phytochemical inhibitors to counteract the cancer promoting effects of VEGFA and TGFBR2 has gained considerable momentum as very few drugs are available. In this study, we employed computational techniques to identify potential VEGFA and TGFBR2 inhibitors from 101 phytochemicals. The Spirosolane, Withaferin A & Silybin B for VEGFA and Ginkgetin, Hesperedin for TGFBR2 were identified as promising candidates as indicated by their favorable docking scores and robust energy profiles. Furthermore, Absorption, Distribution, Metabolism, Excretion, and Toxicity (ADMET) profile indicating that Spirosolane and Ginkgetin exhibit favorable drug‐like properties and medicinal chemistry characteristics, suggesting their potential as novel drug candidates. The Density Functional Theory (DFT) studies revealed that these compounds exhibited the highest electrophilicity index for occupied molecular orbitals, basicity, and dipole moment, all contributing to their stability and strong binding affinity at the active site. To advance this line of research, additional experimental validation is warranted to facilitate the development of highly selective and effective VEGFA and TGFBR2 inhibitors.

Decreased sympathetic nerve activity in young hypertensive rats reared by normotensive mothers

AimsEarly postnatal development can be significantly compromised by changes in factors provided by the mother, leading to increased vulnerability to hypertension in her offspring. TGR(mRen-2)27 (TGR) mothers, characterised by an overactivated renin-angiotensin system, exhibit altered ion composition in their breast milk. Therefore, we aimed to analyse the impact of cross-fostering on cardiovascular parameters in hypertensive TGR and normotensive Hannover Sprague-Dawley (HanSD) offspring. Materials and methodsWe measured cardiovascular parameters in 5- to 10-week-old male offspring by telemetry. The expression of proteins related to vascular function was assessed by western blotting in the aortic samples obtained from 6- to 12-week-old male offspring. Plasma renin activity and plasma angiotensin II (Ang II) levels were evaluated by radioimmunoassay (RIA). Key findingsThe development of hypertension was in TGR accompanied by increased low-to-high frequency ratio (LF/HF; a marker of sympathovagal balance; 0.51 ± 0.16 in week 10). Furthermore, TGR exhibited increased aortic expression of mineralocorticoid receptor (MR; p < 0.05) and transforming growth factor beta type 1 (TGF-β1; p = 0.002) compared to HanSD offspring. Fostering significantly decreased sympathovagal balance (0.23 ± 0.10 in week 10) and, transiently, plasma Ang II levels and MR expression in TGR offspring reared by HanSD mothers. SignificanceThese findings highlight the importance of understanding the complex interplay between early life experiences, maternal factors, and later cardiovascular function. Understanding the mechanisms behind the observed effects may help to identify potential interventions to prevent the development of hypertension later in life.

Activation of immune evasion machinery is a part of the process of malignant transformation of human cells

Malignant transformation of human cells is associated with their re-programming which results in uncontrolled proliferation and in the same time biochemical activation of immunosuppressive pathways which form cancer immune evasion machinery. However, there is no conceptual understanding of whether immune evasion machinery pathways and expression of immune checkpoint proteins form a part of the process of malignant transformation or if they are triggered by T lymphocytes and natural killers (NK) attempting to attack cells which are undergoing or already underwent malignant transformation. To address this fundamental question, we performed experimental malignant transformation of BEAS-2B human bronchial epithelium cells and RC-124 non-malignant human kidney epithelial cells using bracken extracts containing carcinogenic alkaloid called ptaquiloside. This transformation led to a significant upregulation of cell proliferation velocity and in the same time led to a significant upregulation in expression of key immune checkpoint proteins – galectin-9, programmed death ligand 1 (PD-L1), indoleamine 2,3-dioxygenase (IDO1). Their increased expression levels were in line with upregulation of the levels and activities of HIF-1 transcription complex and transforming growth factor beta type 1 (TGF-β)-Smad3 signalling pathway. When co-cultured with T cells, transformed epithelial cells displayed much higher and more efficient immune evasion activity compared to original non-transformed cells. Therefore, this work resolved a very important scientific and clinical question and suggested that cancer immune evasion machinery is activated during malignant transformation of human cells regardless the presence of immune cells in microenvironment.

Galectin-3 regulates skeletal muscle loss via TGFβ1-Smad2/3 signaling activation in mice

IntroductionEndogenous glucocorticoids (GCs) played a pivotal role in the pathogenesis of skeletal muscle loss. However, to date, the underlying molecular mechanisms underlying are not yet fully understood. Galectin-3 (Gal-3) is a member of a beta-galactoside-binding animal lectins, consistently associated with inflammation and fibrosis in the pathogenesis of various disease states. The present study aimed to explore the role of Gal-3 in GC-induced skeletal muscle loss.Material and methodsMyogenic differentiation capacity was detected after in vitro Gal-3 knockdown (KD) or in vivo administration of Gal-3 inhibitor. The activation of transforming growth factor type beta 1 (TGFβ1) and Smad2/3 signaling pathways was determined by western blot, co-immunoprecipitation, and immunofluorescence.ResultsGal-3 was up-regulated during dexamethasone (Dex) administration in mice. In the established GC-induced muscle loss model, Gal-3 inhibition recovered grip strength and muscle mass. In vitro, Gal-3 KD promoted the myogenic differentiation capacity of C2C12 myoblasts, and prevented the reduction of fully differentiated myotubes. Gal-3 results in overexpression of TGFβ1 and TGFβR-I, which affects Smad2, 3 phosphorylation and subsequently mediates skeletal muscle reduction by activating the Smad2/3 signaling pathways.ConclusionsThe present study demonstrated that Dex elevated Gal-3 levels in skeletal muscle. Gal-3 facilitates the activation of TGFβ1-Smad2/3 signaling pathways in myoblasts, and contributes to myogenesis inhibition and skeletal muscle loss. This study raises awareness about the follow-up of patients receiving GC therapy. Further, inhibition of Gal-3 provides a possible therapeutic strategy for skeletal muscle loss.

49P Impact of cyclin D1/TβRII cells in the sensitivity to tamoxifen in hormone receptor-positive breast cancer

It is well established that cyclin D1 is an important component of estrogen regulation. We have previously reported that the transforming growth factor beta type I receptor (TβRI) is related to tamoxifen-sensitive tumors. In addition to TβRI, the type 2 receptor (TβRII) plays a key role in the regulation of TGF-β signaling.The aim of the present study was to explore the predictive value of cyclin D1/TβRII expression for the tamoxifen response in patients with hormone receptor-positive breast cancer.

Inhibiting TGF-β to increase response rates to chemoradiotherapy in rectal cancer

With the rapid increase in the incidence of rectal cancer in young adults, 1 Bailey CE Hu CY You YN et al. Increasing disparities in the age-related incidences of colon and rectal cancers in the United States, 1975–2010. JAMA Surg. 2015; 150: 17-22 Crossref PubMed Scopus (528) Google Scholar interest in organ preservation as an oncologically safe 2 Garcia-Aguilar J Patil S Gollub MJ et al. Organ preservation in patients with rectal adenocarcinoma treated with total neoadjuvant therapy. J Clin Oncol. 2022; (published online April 28.)https://doi.org/10.1200/JCO.22.00032 Crossref PubMed Scopus (28) Google Scholar , 3 Smith JJ Strombom P Chow OS et al. Assessment of a watch-and-wait strategy for rectal cancer in patients with a complete response after neoadjuvant therapy. JAMA Oncol. 2019; 5: e185896 Crossref PubMed Scopus (208) Google Scholar and cost-effective strategy to improve patients' quality of life has also been growing. 4 Miller JA Wang H Chang DT Pollom EL Cost-effectiveness and quality-adjusted survival of watch and wait after complete response to chemoradiotherapy for rectal cancer. J Natl Cancer Inst. 2020; 112: 792-801 Crossref PubMed Scopus (12) Google Scholar Studies evaluating novel radiosensitisers in rectal cancer are receiving attention as a consequence of the need to develop personalised treatment strategies to increase response rates while also facilitating organ preservation. The TGF-β pathway has intrigued scientists and clinicians for decades, 5 Barnard JA Beauchamp RD Coffey RJ Moses HL Regulation of intestinal epithelial cell growth by transforming growth factor type beta. Proc Natl Acad Sci USA. 1989; 86: 1578-1582 Crossref PubMed Scopus (316) Google Scholar and targeting this complex signalling network has now been implicated as a potential way to improve response to radiotherapy. 6 Batlle E Massagué J Transforming growth factor-β signaling in immunity and cancer. Immunity. 2019; 50: 924-940 Summary Full Text Full Text PDF PubMed Scopus (781) Google Scholar However, decades of research into TGF-β inhibitors—both as a monotherapy and in combination with radiotherapy—have shown mixed clinical efficacy, probably due to the complex biology of TGF-β and the context-dependent effects of its inhibition. 6 Batlle E Massagué J Transforming growth factor-β signaling in immunity and cancer. Immunity. 2019; 50: 924-940 Summary Full Text Full Text PDF PubMed Scopus (781) Google Scholar , 7 Andarawewa KL Paupert J Pal A Barcellos-Hoff MH New rationales for using TGFbeta inhibitors in radiotherapy. Int J Radiat Biol. 2007; 83: 803-811 Crossref PubMed Scopus (58) Google Scholar Chest CT scan plus x-ray versus chest x-ray for the follow-up of completely resected non-small-cell lung cancer (IFCT-0302): a multicentre, open-label, randomised, phase 3 trialThe addition of thoracic CT scans during follow-up, which included clinic visits and chest x-rays after surgery, did not result in longer survival among patients with NSCLC. However, it did enable the detection of more cases of early recurrence and second primary lung cancer, which are more amenable to curative-intent treatment, supporting the use of CT-based follow-up, especially in countries where lung cancer screening is already implemented, alongside with other supportive measures. Full-Text PDF

"Management of hospitalised adults with coronavirus disease 2019 (COVID-19): a European Respiratory Society living guideline." James D. Chalmers, Megan L. Crichton, Pieter C. Goeminne, et al. Eur Respir J 2021; 57: 2100048.

<h3>Background</h3> Cancers affect a large number of people worldwide and sometimes have mortality rates of over 90% (pancreatic ductal adenocarcinoma (PDAC), high grade glioblastomas). Currently, immunotherapeutic strategies fail in number of cancer cases due to a lack of understanding of the molecular mechanisms operated by these cancers to evade immune surveillance. Recent evidence demonstrated that the protein galectin-9 is highly expressed in majority of cancer cells. Importantly, this protein is a crucial part of the immune evasion machinery operated by human malignant tumours. Understanding the biochemistry of this immune evasion machinery is required for designing highly efficient and affordable personalised targeted immunotherapy of human cancers. <h3>Materials and Methods</h3> We used, glioblastoma, PDAC, breast and renal cancer cell lines as cancer cell models. Jurkat T lymphocytes, TALL-104 cytotoxic T cells and primary human and mouse CD3-positive T lymphocytes were employed for immune evasion studies. C57 BL16 mice were used for <i>in vivo</i> studies. Western blot analysis, on-cell and in-cell Western, ELISA, qRT-PCR, chromatin immunoprecipitation (ChIP) assays as well as a variety of biochemical tests were used to conduct the studies. <h3>Results</h3> We found that T cells trigger galectin-9 expression and secretion (in most cases requires Tim-3 protein (T cell immunoglobulin and mucine domain containing protein 3)) in a variety of human cancer cells including but not limited to glioblastoma, PDAC, breast and renal cancers. Galectin-9 triggers exhaustion and often death of cytotoxic T cells. Other immune checkpoint proteins including VISTA (V-domain Ig-containing suppressor of T cell activation) and PD-L1 (ligand of programmed cell death protein 1) promote apoptotic death of cytotoxic T cells induced by galectin-9. Activity of T helpers, required to induce cytotoxic T lymphocytes is also suppressed by galectin-9 in cooperation with VISTA and PD-L1. We discovered that expression of all these immune checkpoint proteins is differentially controlled by the autocrine transforming growth factor beta type 1 (TGF-β)-Smad3 pathway as well as several types of interferons. <h3>Conclusions</h3> The immune evasion machinery triggered by TGF-β-Smad3 and interferon pathways and operated by galectin-9, Tim-3, VISTA and PD-L1 should be considered as a potential target for personalised immunotherapy of cancer. Importantly, blood and tumour sample tests could help mapping the exact immune evasion networks employed by each tumour and the pathways responsible to produce respective immune checkpoint proteins. As such, personalised targeted immunotherapy could be adapted. Further investigations required to map the immune evasion machinery for each type of cancer and design the best targets for personalised immunotherapy of human malignancies. <h3>Disclosure Information</h3> <b>V.V. Sumbayev:</b> None. <b>I. Yasinska:</b> None. <b>S. Schlichtner:</b> None. <b>G.S. Lall:</b> None. <b>E. Fasler-Kan:</b> None. <b>B.F. Gibbs:</b> None.

Study of the negative regulation of transforming growth factor beta type II receptor to inhibit the occurrence and development of liver fibrosis with miR-217

Objective: To observe the effect of miR-217 on angiotensin II (AngII)-induced hepatic stellate cells (HSCs) activation, and carbon tetrachloride (CCl4)-induced overexpression in mice, so as to clarify miR-217 role in liver fibrosis. Methods: HSCs were stimulated with AngⅡ and the changes condition in the expression level of miR-217 were detected. HSCs were divided into control group, AngII-treated group and AngⅡ+miR-217-treated group. The expression levels of alpha-smooth muscle actin, fibroblast-specific protein 1 and collagen Ⅰ (Collagen Ⅰ) in each group were detected. The target gene of mir-217 was screened and verified by Targetscan and Dual luciferase gene reporter assay. Real-time quantitative PCR and Western blot were used to detect the effect of miR-217 on the expression level of transforming growth factor beta type Ⅱ receptor (TGFBR2). A CCl4-induced mouse liver fibrosis model was constructed. Masson staining and Sirius red staining were used to detect the effect of miR-217 overexpression on the progression of liver fibrosis in CCl4 mice. Data of two groups were compared using t-test. Data of multiple groups were statistically analyzed with one-way ANOVA. Results: The expression level of miR-217 was downregulated by AngⅡ-stimulated HSC cells. The expression levels of α-smooth muscle actin, fibroblast-specific protein 1 and Collagen Ⅰ induced by AngⅡ was inhibited by miR-217 mimics transfection. The 3'-UTR of TGFBR2 had specifically bind miR-217. The mRNA and protein expression levels of TGFBR2 was inhibited with miR-217 mimics transfection in HSCs. The overexpression of miR-217 had inhibited the expression levels of Collagen Ⅰ and Ⅲ in CCl4 mice and alleviated the progression of liver fibrosis . Conclusion: miR-217 regulates liver fibrosis by targeting TGFBR2, inhibits AngII-induced HSC activation, and slows down the process of liver fibrosis in CCl4 mice, suggesting that miR-217 may have an inhibitory effect on liver fibrosis.

Design, synthesis, in silico and antibacterial evaluation of curcumin derivatives loaded nanofiber as potential wound healing agents

ObjectiveDrug resistance can result in mortality and morbidity due to treatment failures. Therefore, researchers are actively involved in the urgent development of new antibacterial agents which are more effective against drug-resistant bacteria. This study is aimed to prepare curcumin derivatives (CD) nanofiber and test its antibacterial activity by disk diffusion test and evaluate their role in wound healing when potentially binds with transforming growth factor-beta type 1 kinase domain (TGF-β) and Glycogen synthase kinase-3 beta (GSK3-β). MethodMolecular docking and molecular dynamic have been used to select the most potent dual-action molecules on both TGF-β and GSK3-β, most active 2 compounds have been synthesized and characterized by 1H and 13C (NMR), IR spectroscopy, Electrospinning of CD in ethanol were studied by adding Poly vinyl alcohol (PVA) as a polymer. The properties of the CD\\PVA solutions measured, morphology of electrospun CD\\PVA was tested by Scanning Electrode Microscope (SEM), antibacterial activity of electrospun nanofiber against Staphylococcus aureus (S. aureus) was tested by disk diffusion method.Results and Conclusions: CD nanofiber were synthesized successfully. The results showed that CD were successfully synthesized with good yield (72% and 78%) for compound1 and 2 respectively, Disc diffusion antibacterial test demonstrated an increased inhibition zone from 0.5 to 5.0 mg loaded CD with the largest inhibition zone of 8.5 ± 0.71 and 9.67 ± 0.29 mm for compound1 and 2 respectively. Hence, it can be concluded that these CD\\PVA could be a new source in designing new antibacterial wound dressing for diabetic patients.

Expression of the Immune Checkpoint Protein VISTA Is Differentially Regulated by the TGF-β1 - Smad3 Signaling Pathway in Rapidly Proliferating Human Cells and T Lymphocytes.

Immune checkpoint proteins play crucial roles in human embryonic development but are also used by cancer cells to escape immune surveillance. These proteins and biochemical pathways associated with them form a complex machinery capable of blocking the ability of cytotoxic immune lymphoid cells to attack cancer cells and, ultimately, to fully suppress anti-tumor immunity. One of the more recently discovered immune checkpoint proteins is V-domain Ig-containing suppressor of T cell activation (VISTA), which plays a crucial role in anti-cancer immune evasion pathways. The biochemical mechanisms underlying regulation of VISTA expression remain unknown. Here, we report for the first time that VISTA expression is controlled by the transforming growth factor beta type 1 (TGF-β)-Smad3 signaling pathway. However, in T lymphocytes, we found that VISTA expression was differentially regulated by TGF-β depending on their immune profile. Taken together, our results demonstrate the differential biochemical control of VISTA expression in human T cells and various types of rapidly proliferating cells, including cancer cells, fetal cells and keratinocytes.

MicroRNA-210–5p alleviates cardiac fibrosis via targeting transforming growth factor-beta type I receptor in rats on high sodium chloride (NaCl)-based diet

The present study was designed to explore whether high sodium chloride (NaCl)-based diet (HSD) caused cardiac fibrosis regardless of blood pressure in Sprague-Dawley (SD) rats, and to further determine the effects and the underlying mechanisms of microRNA (miR)-210–5p on HSD-induced cardiac fibrosis in rats or NaCl-induced cardiac fibroblast activation in neonatal rat cardiac fibroblasts (NRCFs). The SD rats received 8% HSD, and NRCFs were treated with NaCl. The levels of collagen I, alpha-smooth muscle actin (α-SMA) and transforming growth factor-beta 1 (TGF-β1) were increased in the heart of hypertension (HTN), hypertension-prone (HP) and hypertension-resistant (HR) rats on HSD in vivo. NaCl increased the levels of collagen I, α-SMA and TGF-β1 in NRCFs in vitro. The level of miR-210–5p was reduced in both NBD-induced rats’ hearts and NaCl-treated NRCFs, which was consistent with the results of miR high-throughput sequencing in NRCFs. The HSD or NaCl-induced increases of collagen I, α-SMA and TGF-β1 were inhibited by miR-210–5p agomiR in vitro and in vivo, respectively. miR-210–5p antagomiR could mimic the pathological effects of NaCl in NRCFS. Bioinformatics analysis and luciferase reporter assays demonstrated that TGF-β type I receptor (TGFBR1) was a direct target gene of miR-210–5p. These results indicated that HSD resulted in cardiac fibrosis regardless of blood pressure. The upregulation of miR-210–5p could attenuate cardiac fibroblast activation in NRCFS via targeting TGFBR1. Thus, upregulating miR-210–5p might be a strategy for the treatment of cardiac fibrosis.

High Mobility Group Box 1 (HMGB1) Induces Toll-Like Receptor 4-Mediated Production of the Immunosuppressive Protein Galectin-9 in Human Cancer Cells.

High mobility group box 1 (HMGB1) is a non-histone protein which is predominantly localised in the cell nucleus. However, stressed, dying, injured or dead cells can release this protein into the extracellular matrix passively. In addition, HMGB1 release was observed in cancer and immune cells where this process can be triggered by various endogenous as well as exogenous stimuli. Importantly, released HMGB1 acts as a so-called “danger signal” and could impact on the ability of cancer cells to escape host immune surveillance. However, the molecular mechanisms underlying the functional role of HMGB1 in determining the capability of human cancer cells to evade immune attack remain unclear. Here we report that the involvement of HMGB1 in anti-cancer immune evasion is determined by Toll-like receptor (TLR) 4, which recognises HMGB1 as a ligand. We found that HGMB1 induces TLR4-mediated production of transforming growth factor beta type 1 (TGF-β), displaying autocrine/paracrine activities. TGF-β induces production of the immunosuppressive protein galectin-9 in cancer cells. In TLR4-positive cancer cells, HMGB1 triggers the formation of an autocrine loop which induces galectin-9 expression. In malignant cells lacking TLR4, the same effect could be triggered by HMGB1 indirectly through TLR4-expressing myeloid cells present in the tumour microenvironment (e. g. tumour-associated macrophages).

Ectopic FOXP3 Expression in Combination with TGF-β1 and IL-2 Stimulation Generates Limited Suppressive Function in Human Primary Activated Thymocytes Ex Vivo.

Regulatory T cells (Tregs), which are characterized by the expression of the transcription factor forkhead box P3 (FOXP3), are the main immune cells that induce tolerance and are regulators of immune homeostasis. Natural Treg cells (nTregs), described as CD4+CD25+FOXP3+, are generated in the thymus via activation and cytokine signaling. Transforming growth factor beta type 1 (TGF-β1) is pivotal to the generation of the nTreg lineage, its maintenance in the thymus, and to generating induced Treg cells (iTregs) in the periphery or in vitro arising from conventional T cells (Tconvs). Here, we tested whether TGF-β1 treatment, associated with interleukin-2 (IL-2) and CD3/CD28 stimulation, could generate functional Treg-like cells from human thymocytes in vitro, as it does from Tconvs. Additionally, we genetically manipulated the cells for ectopic FOXP3 expression, along with the TGF-β1 treatment. We demonstrated that TGF-β1 and ectopic FOXP3, combined with IL-2 and through CD3/CD28 activation, transformed human thymocytes into cells that expressed high levels of Treg-associated markers. However, these cells also presented a lack of homogeneous suppressive function and an unstable proinflammatory cytokine profile. Therefore, thymocyte-derived cells, activated with the same stimuli as Tconvs, were not an appropriate alternative for inducing cells with a Treg-like phenotype and function.

MicroRNA-34a Promotes EMT and Liver Fibrosis in Primary Biliary Cholangitis by Regulating TGF-β1/smad Pathway.

Background and Aims Primary biliary cholangitis (PBC) is an autoimmune cholestatic liver disease. We found microRNA-34a (miR-34a), as the downstream gene of p53, was overexpressed in some of fibrogenic diseases. In this study, we sought to explore whether miR-34a plays a role in the fibrosis of PBC. Methods The peripheral blood of PBC patients and controls was collected to analyze the level of miR-34a. Human intrahepatic biliary epithelial cells (HIBEC) were cultured. The expression of miR-34a was regulated by miR-34a mimics and inhibitor. The biomarkers of epithelium-mesenchymal transition (EMT), fibrogenesis, inflammation, and transforming growth factor- (TGF-) β1/smad pathway were analyzed. Results We found that miR-34a was overexpressed in the peripheral blood in PBC patients. In vitro, overexpressed miR-34a increased the EMT and fibrogenesis activity of HIBEC. Transforming growth factor-beta type 1 receptor (TβR1), TGF-β1, and p-smad2/3 were upregulated by miR-34a. Inflammatory factors such as IL-6 and IL-17 were also upregulated. Finally, we showed that miR-34a promoted EMT and liver fibrosis in PBC by targeting the TGF-β1/smad pathway antagonist transforming growth factor-beta-induced factor homeobox 2 (TGIF2). Conclusions Our findings show that miR-34a plays an important role in the EMT and fibrosis of PBC through the TGF-β1/smad pathway by targeting TGIF2. This study suggests that miR-34a may be a new marker of fibrogenesis in PBC. Inhibition of miR-34a may be a promising strategy in treating PBC and improving the prognosis of the disease.

Suppressing long noncoding RNA OGRU ameliorates diabetic retinopathy by inhibition of oxidative stress and inflammation via miR-320/USP14 axis

Long noncoding RNAs (lncRNAs) are important regulators in various diseases including diabetic retinopathy (DR). In this study, DR patients exhibited significantly increased expression of serum LncRNA-OGRU compared with normal individuals. Streptozotocin (STZ)-challenged rats with DR also had higher OGRU expression in retinas than that of the control group, which was confirmed in Müller cells upon high glucose (HG) stimulation. OGRU knockdown remarkably decreased vascular endothelial growth factor (VEGF) and transforming growth factor-β1 (TGF-β1) expression in HG-incubated Müller cells. HG-induced inflammatory response and oxidative stress in vitro were markedly mitigated by OGRU knockdown through restraining IκBɑ/nuclear factor kappa beta (NF-κB) and improving nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathways, respectively. Further studies indicated that OGRU suppression greatly restored miR-320 expression, and a negative correlation between them was detected in DR patients. We also found that miR-320 over-expression considerably restrained TGF-β1 signaling, and hindered inflammation and reactive oxygen species (ROS) production in HG-stimulated Müller cells. Additionally, OGRU knockdown or miR-320 over-expression could dramatically down-regulate ubiquitin-specific peptidase 14 (USP14) expression levels in HG-incubated Müller cells, and miR-320 could directly target USP14. Notably, OGRU/miR-320 axis-mediated TGF-β1 signaling, inflammation and ROS were largely dependent on USP14. Intriguingly, our results showed that USP14 directly interacted with transforming growth factor-beta type 1 receptor (TβR1), and impeded TβR1 ubiquitination and degradation. Furthermore, USP14 could also facilitate IκBɑ deubiquitination and degradation, exacerbating IκBɑ phosphorylation and NF-κB activation. Finally, our in vivo studies confirmed that OGRU knockdown considerably ameliorated DR progression in STZ-challenged rats through mediating the mechanisms observed in vitro. Collectively, these findings implicated that LncRNA-OGRU mediated DR progression through competing for miR-320 to regulate USP14 expression, and thus LncRNA-OGRU/miR-320/USP14 axis may be considered as a therapeutic target for DR treatment.

Piperlongumine inhibits the growth of non-small cell lung cancer cells via the miR-34b-3p/TGFBR1 pathway

BackgroundNon-small cell lung cancer is a common type of lung cancer. Piperlongumine (PL), which is extracted from the roots of piperaceae plant, long pepper, and peppercorn, is an alkaloid amide that inhibits tumor growth and metastasis. However, whether it affects lung cancer cells remains unclear.MethodsWe assessed the effects of PL on the proliferation and apoptosis of A549 and H1299 NSCLC cell lines.ResultsPL was mildly toxic to normal human bronchial epithelial cells and significantly suppressed growth and facilitated apoptosis of A549 and H1299 cells. It also upregulated microRNA (miR)-34b-3p and downregulated the transforming growth factor beta type I receptor (TGFBR1). The dual-luciferase reporter assay showed that TGFBR1 is a target gene of miR-34b-3p. Silencing of miR-34b-3p or overexpression of TGFBR1 partially attenuated the effects of PL on A549 and H1299 cells.ConclusionsPL inhibits proliferation and induces apoptosis of A549 and H1299 cells by upregulating miR-34b-3p and modulating TGFBR1 signaling pathway.

Transforming growth factor beta type 1 (TGF-β) and hypoxia-inducible factor 1 (HIF-1) transcription complex as master regulators of the immunosuppressive protein galectin-9 expression in human cancer and embryonic cells.

Galectin-9 is one of the key proteins employed by a variety of human malignancies to suppress anti-cancer activities of cytotoxic lymphoid cells and thus escape immune surveillance. Human cancer cells in most cases express higher levels of galectin-9 compared to non-transformed cells. However, the biochemical mechanisms underlying this phenomenon remain unclear.Here we report for the first time that in human cancer as well as embryonic cells, the transcription factors hypoxia-inducible factor 1 (HIF-1) and activator protein 1 (AP-1) are involved in upregulation of transforming growth factor beta 1 (TGF-β1) expression, leading to activation of the transcription factor Smad3 through autocrine action. This process triggers upregulation of galectin-9 expression in both malignant (mainly in breast and colorectal cancer as well as acute myeloid leukaemia (AML)) and embryonic cells. The effect, however, was not observed in mature non-transformed human cells. TGF-β1-activated Smad3 therefore displays differential behaviour in human cancer and embryonic vs non-malignant cells. This study uncovered a self-supporting biochemical mechanism underlying high levels of galectin-9 expression operated by the human cancer and embryonic cells employed in our investigations. Our results suggest the possibility of using the TGF-β1 signalling pathway as a potential highly efficient target for cancer immunotherapy.

Based on G-Series Mouse TH17 Array Study the Effect of Fluoride on C2C12 Cells Cytokines Expression.

C2C12 cells were cultured on medium containing fluoride (0, 1, and 2.5 mmol/L) for 48 h to investigate the effect of excessive fluoride on T helper 17 (Th17)-related cytokine expression profile in skeletal muscle cells, and the culture supernatant was collected and subjected for the detection of 18 cytokines via Th17 array. Results showed that compared with the control group, no differential expression proteins (DEPs) were found in the 1 mmol/L fluoride group; however, eight DEPs were upregulated in the 2.5 mmol/L fluoride group, including macrophage inflammatory protein-3α (MIP-3α), interleukin-21 (IL-21), IL-13, IL-17F, IL-28A, transforming growth factor type beta 1 (TGF-β1), IL-23, and IL-17A. In addition, five DEPs (MIP-3α, IL-13, IL-21, TGF-β1, and IL-17F) were upregulated in the 2.5 mmol/L fluoride group compared with the 1 mmol/L fluoride group. Gene ontology analysis revealed that the positive regulation of cytokine production, cytokine activity, receptor ligand activity, and cytokine receptor binding accounted for high percent of DEPs present. Kyoto Encyclopedia of Genes and Genomes analysis showed that these DEPs primarily involved 12 pathways enriched in the cytokine-cytokine receptor interaction and IL-17 signaling pathway after 2.5 mmol/L fluoride treatment. The results indicated that fluoride might induce cytotoxicity by disturbing Th17-related cytokine expression.

Transcriptomic approach predicts a major role for transforming growth factor beta type 1 pathway in L-Dopa-induced dyskinesia in parkinsonian rats.

Dyskinesia induced by long-term L-Dopa (LID) therapy in Parkinson disease is associated with altered striatal function whose molecular bases remain unclear. Here, a transcriptomic approach was applied for comprehensive analysis of distinctively regulated genes in striatal tissue, their specific pathways, and functional- and disease-associated networks in a rodent model of LID. This approach has identified transforming growth factor beta type 1 (TGFβ1) as a highly upregulated gene in dyskinetic animals. TGFβ1 pathway is a top aberrantly regulated pathway in the striatum following LID development based on differentially expressed genes (> 1.5 fold change and P < 0.05). The induction of TGFβ1 pathway specific genes, TGFβ1, INHBA, AMHR2 and PMEPA1 was also associated with regulation of NPTX2, PDP1, SCG2, SYNPR, TAC1, TH, TNNT1 genes. Transcriptional network and upstream regulator analyses have identified AKT-centered functional and ERK-centered disease networks revealing the association of TGFβ1, IL-1β and TNFα with LID development. Therefore, results support that TGFβ1 pathway is a major contributor to the pathogenic mechanisms of LID.

Docking-based virtual screening of T\u03b2R1 inhibitors: evaluation of pose prediction and scoring functions

To improve the reliability of virtual screening for transforming growth factor-beta type 1 receptor (TβR1) inhibitors, 2 docking methods and 11 scoring functions in Discovery Studio software were evaluated and validated in this study. LibDock and CDOCKER protocols were performed on a test set of 24 TβR1 protein–ligand complexes. Based on the root-mean-square deviation (RMSD) values (in Å) between the docking poses and co-crystal conformations, the CDOCKER protocol can be efficiently applied to obtain more accurate dockings in medium-size virtual screening experiments of TβR1, with a successful docking rate of 95%. A dataset including 281 known active and 8677 inactive ligands was used to determine the best scoring function. The receiver operating characteristic (ROC) curves were used to compare the performance of scoring functions in attributing best scores to active than inactive ligands. The results show that Ludi 1, PMF, Ludi 2, Ludi 3, PMF04, PLP1, PLP2, LigScore2, Jain and LigScore1 are better scoring functions than the random distribution model, with AUC of 0.864, 0.856, 0.842, 0.812, 0.776, 0.774, 0.769, 0.762, 0.697 and 0.660, respectively. Based on the pairwise comparison of ROC curves, Ludi 1 and PMF were chosen as the best scoring functions for virtual screening of TβR1 inhibitors. Further enrichment factors (EF) analysis also supports PMF and Ludi 1 as the top two scoring functions.