Transforming Growth Factor Beta Protein Research Articles

Transforming Growth Factor-β/Smad Signaling Inhibits Melanoma Cancer Stem Cell Self-Renewal, Tumor Formation and Metastasis.

The secreted protein transforming growth factor-beta (TGFβ) plays essential roles, ranging from cell growth regulation and cell differentiation in both normal and cancer cells. In melanoma, TGFβ acts as a potent tumor suppressor in melanoma by blocking cell cycle progression and inducing apoptosis. In the present study, we found TGFβ to regulate cancer stemness in melanoma through the Smad signaling pathway. We discovered that TGFβ/Smad signaling inhibits melanosphere formation in multiple melanoma cell lines and reduces expression of the CD133+ cancer stem cell subpopulation in a Smad3-dependent manner. Using preclinical models of melanoma, we further showed that preventing Smad3/4 signaling, by means of CRISPR knockouts, promoted both tumorigenesis and lung metastasis in vivo. Collectively, our results define new functions for the TGFβ/Smad signaling axis in melanoma stem-cell maintenance and open avenues for new therapeutic approaches to this disease.

A-055 Clinical Performance Evaluation of the ADVIA Centaur AMH Assay

Abstract Background Anti-Müllerian hormone (AMH) is a transforming growth factor beta protein synthesized by granulosa cells of small growing follicles in ovaries in females and by Sertoli cells of the testes in males. AMH measurements are increasingly being used in the assessment of ovarian reserve, prediction of response to controlled ovarian stimulation in women undergoing assisted reproductive technologies, as an aid in the diagnostic evaluation of polycystic ovary syndrome, and prediction of menopause. The objective of this study was to evaluate the clinical performance of the ADVIA Centaur® AMH assay in the assessment of ovarian reserve and compare performance to the Beckman ACCESS 2 AMH assay. In addition, an exploratory analysis evaluated ADVIA Centaur AMH assay performance in distinguishing women with expected poor ovarian response per published criteria. Methods Clinical specimens were collected from 577 women at 11 U.S. fertility centers. Women enrolled were 22–45 years old and possessed both ovaries. They underwent blood draw and transvaginal ultrasound to measure the antral follicle count on days 2–4 of menstrual cycle. Women with BMI ≥40; pregnancy within 3 months; on contraceptives, hormonal medications; undergoing treatment for malignancy, ovarian abnormalities, surgery in the past 6 months; or with endocrine or metabolic disorders (including PCOS) were excluded. Results Results were considered positive (high ovarian reserve) if the AMH concentration was >1.77 ng/mL and AFC was >15 and negative (normal-to-diminished ovarian reserve) if the AMH concentration was ≤1.77 ng/mL and AFC was ≤15. The estimated sensitivity, specificity, and two-sided 95% Wilson score confidence intervals (CI) of each parameter were determined to be 90.76% (86.97–93.53) and 52.19% (46.29–58.03) using the ADVIA Centaur AMH assay, and 90.10% (86.22–92.98) and 52.19% (46.29–58.03) with the ACCESS 2 AMH assay, respectively.A secondary analysis was completed to investigate poor ovarian response (POR) according to the Bologna criteria, published by the ESHRE. These criteria define poor ovarian response as an AMH measurement of less than 1.1 ng/mL and AFC of less than 7. Of the 577 women included in the study, 32 women had an antral follicle count less than 7, of whom 26 also had a AMH concentration of less than 1.1 ng/mL. Of the 545 women with an antral follicle count greater than or equal to 7, 480 also had AMH measurements of greater than or equal to 1.1 ng/mL. Using the Bologna criteria, the ADVIA Centaur AMH assay performs with a sensitivity of 81.25% and specificity of 88.07%. Conclusions In this clinical population, the sensitivity and specificity of AMH assays on the ADVIA Centaur XP and Beckman ACCESS 2 systems were almost identical when using an AMH cutoff ≤1.77 ng/mL and AFC ≤15 to identify normal-to-diminished ovarian reserve. Using published criteria for POR resulted in a sensitivity and specificity of 81.25% and 88.07% for the ADVIA Centaur AMH assay. Not available for sale in the U.S. Future availability cannot be guaranteed. Product availability varies by county. HOOD#05162003331923

Relationship between TGF-Beta 1 rs1800469 Gene Polymorphism with Degree of Tumor Invasion of Colorectal Cancer at RSUP Prof. Dr. I.G.N.G. Ngoerah Denpasar Bali

The incidence of cancer, including colorectal cancer (CRC), increases with age. The Transforming Growth Factor-Beta 1 (TGF-B1) rs1800469 gene polymorphism is associated with the pathogenesis of CRC. The T allele is associated with increased transcriptional activity compared to the C allele, meaning that individuals with TT homozygotes will produce higher TGF-B1 protein concentrations. This study aimed to analyze the relationship between the TGF-B1 rs1800469 gene polymorphism and degree of tumor invasion of CRC in Bali, especially at the RSUP Prof. Dr. I.G.N.G. Ngoerah Denpasar. This hospital study was a cross-sectional study using formalin-fixed-paraffin embedded (FFPE) samples from CRC patients, stored in the Department of Pathology Prof. Dr. I.G.N.G. Ngoerah Denpasar Bali. Identification of TGF-B1 rs1800469 polymorphism gene was performed using polymerase chain reaction (PCR) and sequencing methods. Data on age, sex, histopathological grading, degree of tumor invasion, pathological staging N and tumor location were recorded from the patient's medical record, carried out descriptive analysis and hypothesis testing of bivariate Chi Square analysis. Out of 48 samples, majority was over 50 years old (83.3%), male (54.2%), low grade histopathology (93.8%), adenocarcinoma histological classification (97.9%), high degree of tumor invasion (81.2%), staging pathology N with regional lymph nodes cannot be assessed (93.7%), left tumor location (58.3%) with TGF-B1 rs1800469 gene polymorphism CT genotype of 23 samples (47.9%), followed by TT genotypes of 17 samples (35.4%) and CC genotypes of 8 samples (16.7%). The results of the Chi Square statistical analysis test found no association between the TGF-B1 rs1800469 gene polymorphism in any genotype and degree of tumor invasion of CRC (p=0.688). It can be concluded that there was no significant relationship between the TGF-B1 rs1800469 gene polymorphism and degree of tumor invasion of CRC patients at RSUP Prof. Dr. I.G.N.G. Ngoerah, Denpasar, Bali. Keywords: Colorectal Cancer, Gene Polymorphism, SNP, Tumor Invasion

The role of transforming growth factor beta-1 protein in Escherichia coli secondary infection induced by H9N2 avian influenza virus in chickens

The H9N2 subtype of avian influenza virus (AIV) is common in poultry production. It causes mild clinical signs but rarely leads to poultry mortalities. However, higher mortality can occur in chickens with co-infections, especially avian pathogenic Escherichia coli (APEC), which results in huge economic losses for the poultry industry. Unfortunately, the mechanism of co-infection remains unknown. Our previous studies screened several proteins associated with bacterial adhesion, including transforming growth factor beta-1 (TGF-β1), integrins, cortactin, E-cadherin, vinculin, and fibromodulin. Herein, we investigated the contribution of TGF-β1 to APEC adhesion after H9N2 infection. We first infected H9N2 and APEC in chicken, chicken embryo and DF-1cells, and demonstrated that H9N2 infection promotes APEC adhesion to hosts in vitro and in vivo by plate count method. Through real-time fluorescence quantification and enzyme-linked immunosorbent assay, it was demonstrated that H9N2 infection not only increases TGF-β1 expression but also its activity in a time-dependent manner. Then, through exogenous addition of TGF-β1 and overexpression, we further demonstrated that TGF-β1 can increase the adhesion of endothelial cells to DF-1cells. Furthermore, the capacity of APEC adhesion to DF-1cells was significantly decreased either by adding a TGF-β1 receptor inhibitor or using small interfering RNAs to interfere with the expression of TGF-β1. To sum up, H9N2 infection can promote the upregulation of TGF-β1 and then increase the adhesion ability of APEC. Targeting TGF-β1 and its associated pathway will provide valuable insights into the clinical treatment of E. coli secondary infection induced by H9N2 infection.

Microvascular significance of TGF-β axis activation in COVID-19.

As 2023 approaches, the COVID-19 pandemic has killed millions. While vaccines have been a crucial intervention, only a few effective medications exist for prevention and treatment of COVID-19 in breakthrough cases or in unvaccinated or immunocompromised patients. SARS-CoV-2 displays early and unusual features of micro-thrombosis and immune dysregulation that target endothelial beds of the lungs, skin, and other organs. Notably, anticoagulation improves outcomes in some COVID-19 patients. The protein transforming growth factor-beta (TGF-β1) has constitutive roles in maintaining a healthy microvasculature through its roles in regulating inflammation, clotting, and wound healing. However, after infection (including viral infection) TGF-β1 activation may augment coagulation, cause immune dysregulation, and direct a path toward tissue fibrosis. Dysregulation of TGF-β signaling in immune cells and its localization in areas of microvascular injury are now well-described in COVID-19, and such events may contribute to the acute respiratory distress syndrome and skin micro-thrombosis outcomes frequently seen in severe COVID-19. The high concentration of TGF-β in platelets and in other cells within microvascular thrombi, its ability to activate the clotting cascade and dysregulate immune pathways, and its pro-fibrotic properties all contribute to a unique milieu in the COVID-19 microvasculature. This unique environment allows for propagation of microvascular clotting and immune dysregulation. In this review we summarize the physiological functions of TGF-β and detail the evidence for its effects on the microvasculature in COVID-19. In addition, we explore the potential role of existing TGF-β inhibitors for the prevention and treatment of COVID-19 associated microvascular thrombosis and immune dysregulation.

M2 macrophages mediate fibrotic scar formation in the early stages after cerebral ischemia in rats.

In the central nervous system, the formation of fibrotic scar after injury inhibits axon regeneration and promotes repair. However, the mechanism underlying fibrotic scar formation and regulation remains poorly understood. M2 macrophages regulate fibrotic scar formation after injury to the heart, lung, kidney, and central nervous system. However, it remains to be clarified whether and how M2 macrophages regulate fibrotic scar formation after cerebral ischemia injury. In this study, we found that, in a rat model of cerebral ischemia induced by middle cerebral artery occlusion/reperfusion, fibrosis and macrophage infiltration were apparent in the ischemic core in the early stage of injury (within 14 days of injury). The number of infiltrated macrophages was positively correlated with fibronectin expression. Depletion of circulating monocyte-derived macrophages attenuated fibrotic scar formation. Interleukin 4 (IL4) expression was strongly enhanced in the ischemic cerebral tissues, and IL4-induced M2 macrophage polarization promoted fibrotic scar formation in the ischemic core. In addition, macrophage-conditioned medium directly promoted fibroblast proliferation and the production of extracellular matrix proteins in vitro. Further pharmacological and genetic analyses showed that sonic hedgehog secreted by M2 macrophages promoted fibrogenesis in vitro and in vivo, and that this process was mediated by secretion of the key fibrosis-associated regulatory proteins transforming growth factor beta 1 and matrix metalloproteinase 9. Furthermore, IL4-afforded functional restoration on angiogenesis, cell apoptosis, and infarct volume in the ischemic core of cerebral ischemia rats were markedly impaired by treatment with an sonic hedgehog signaling inhibitor, paralleling the extent of fibrosis. Taken together, our findings show that IL4/sonic hedgehog/transforming growth factor beta 1 signaling targeting macrophages regulates the formation of fibrotic scar and is a potential therapeutic target for ischemic stroke.

Protein signatures of heart failure mortality in the community

Abstract Background Heart failure (HF) is a complex clinical syndrome that affects over 60 million persons worldwide.1 The 5-year survival rate of HF is less than 50% irrespective of ejection fraction (EF) and the limitations of EF in sufficiently phenotyping HF are recognized.2,3 High throughput proteomics assays are an attractive option for precision phenotyping.4 Purpose To explore the proteomic profiles and biomarker pathways associated with death in an HF community cohort. Methods We measured proteomics in plasma specimens collected from an HF cohort (2003–2012) of patients predominantly of European Ancestry (92%). The relative concentrations of circulating plasma proteins were measured using an aptamer-based assay containing 7335 human protein targets. Proteins significantly associated with mortality were selected while adjusting for age, sex, and eGFR. These proteins were employed in clustering analysis and subjected to 10-fold cross-validation. We quantified and corrected the clustering bias via bootstrapping and subsequently validated the results via semi-supervised clustering. Multivariable Cox regression was implemented for the cluster variable while adjusting for the Meta-analysis Global Group in Chronic Heart Failure (MAGGIC) risk score. Additionally, we performed enrichment and pathway analysis of the outcome-associated proteins considering significance thresholds of −log10(P-value) >2, FDR corrected P<0.05 (regulator analysis) and Z-scores of >2 and <−2 as thresholds for activation and inhibition of proteins/pathways. Results From a total of 1,388 patients (mean age 75±13; 52% male), 2 distinct clusters, (HR: 2.04; 95% CI [1.78–2.34]; p-value: <0.0001 independent of MAGGIC) were identified, cluster 1; n=722, mean age 72±14; NYHA class III/IV: 41%/26%; EF: 47%±17 and cluster 2; n=666, mean age 79±11; NYHA class III/IV: 41%/31%, EF: 48%±16 based on the 447 proteins [354 positively and 93 negatively] significantly associated with mortality (1,158 events). Gene-set enrichment analysis of all 447 proteins revealed cell adhesion, extracellular matrix organization, enzyme-linked protein/tyrosine kinase signaling pathways, humoral immune response, tissue development, growth factor signaling, post-translational protein modification, and platelet degranulation as the dominant biological themes. Finally, we found 18 upstream regulators predicted to be driving the expressions of the proteins in the dataset including a predicted outcome-associated protein-Transforming Growth Factor Beta 1 – TGFB1 (p=7.46E-09), Interleukin 1B, IL1B (p=9.40E-08), and oncostatin M, OSM (p=6.61E-05) as the top 3 regulators (predicted activation) based on FDR-corrected statistical significance of P<0.05 (Figure 1) Conclusion We identified distinct clusters based on proteomic profiles associated with HF mortality independently of clinical prognostic makers. Further, we delineated protein regulators driving the major pathways associated with HF survival. Funding Acknowledgement Type of funding sources: Public Institution(s). Main funding source(s): Department of Intramural Research, NIH/NHLBI

Blocking of CA-MSC-induced desmoplasia reprograms the tumor immune microenvironment and enhances cancer immunotherapy

Abstract Cancer associated MSCs (CA-MSCs) are multipotent stromal cells that generate tumor desmoplasia and promote tumor growth. Here, we investigated the impact of CA-MSCs on the ovarian tumor immune microenvironment. In vitro and in vivo analyses confirmed that CA-MSCs secrete numerous cytokines and chemokines, including Ccl2, Cx3cl1 and Tgfb1, that are known to induce the recruitment of monocytes into the tumor and promote macrophage immunosuppressive phenotype. Using a syngeneic and immune responsive murine ovarian tumor model, we found that CA-MSCs restrict the intra-tumoral trafficking of CD8+ T cells and inhibit the response to immune checkpoint inhibitor therapy. Similarly, in patient samples the presence of CA-MSC is inversely correlated with CD8+ T cell infiltration into tumor islets. Single cell RNAseq analyses revealed that tumoral monocytes and macrophages of a-PD-L1 treated-CA-MSCs enriched tumors express high levels of the protein transforming growth factor beta induced (Tgfbi). In ovarian cancer patient samples TGFBI is primarily expressed in the desmoplastic stroma and correlated with poor prognosis. Importantly, we found that hedgehog signaling inhibitor (HHi) therapy reversed CA-MSC driven tumor desmoplasia, reduced the number of tumor-associated myeloid cells and decreased their expression of Tgfbi. These events restored the infiltration and activation of CD8+ T cells into the tumor islets and response to a-PDL1 therapy. We are currently evaluating the role of Tgfbi in the regulation of innate and adaptive immune response. Our findings demonstrated CA-MSCs critically regulate anti-tumor immunity and support HHi as an adjunct to anti-PDL1 therapy to increase repose to immune therapy in ovarian cancer.

Effects of Angiotensin Converting Enzyme 2 and Angiotensin Converting Enzyme Genes on Cardiac Remodeling in Rats with Heart Failure

To investigate the effects of angiotensin converting enzyme 2 and angiotensin converting enzyme genes on the cardiac remodeling process in heart failure rats. 38 rats were randomly divided into sham-operated group (S group), heart failure group (H group) and heart failure exercise group (HE group). An incremental load exercise test was used to determine the maximum running speed of the HE group, while the S and H groups maintained their daily activities. The cardiac structure and function and the pathological changes were compared. Compared with S group, body weight, left ventricular internal diameter at diastole, fractional shortening, left ventricular ejection fraction, myocytes and angiotensin converting enzyme/ angiotensin converting enzyme 2 ratio were decreased while left ventricular weight, left ventricular mass index, ventricular anterior wall diameter at diastole, left ventricular anterior wall diameter at systole, left ventricular posterior wall diameter at diastole, left ventricular posterior wall diameter at systole, atrial natriuretic factor, angiotensin-I, angiotensin-II, messenger RNA and angiotensin converting enzyme, angiotensin converting enzyme 2, transforming growth factor beta protein levels and collagen fibers (i.e. collagen volume fraction) were increased in H group (p<0.05). Compared with H group, left ventricular weight, left ventricular mass index, left ventricular internal diameter at diastole, fractional shortening, left ventricular ejection fraction, myocytes were increased while collagen volume fraction, atrial natriuretic factor, angiotensin-I, angiotensin-II, messenger RNA and angiotensin converting enzyme, angiotensin converting enzyme 2, transforming growth factor beta protein levels were reduced and angiotensin converting enzyme/angiotensin converting enzyme 2 ratio were decreased (p<0.05). Long-term aerobic exercise attenuated myocardial fibrosis and improved cardiac function by inhibiting collagen synthesis and promoting its degradation and cardiac remodeling was inhibited and improved by a mechanism that may be related to partial restoration of angiotensin converting enzyme 2/angiotensin converting enzyme signalling function.

TGFBI modulates tumour hypoxia and promotes breast cancer metastasis.

Breast cancer metastasis is a complex process that depends not only on intrinsic characteristics of metastatic stem cells, but also on the particular microenvironment that supports their growth and modulates the plasticity of the system. In search for microenvironmental factors supporting cancer stem cell (CSC) growth and tumour progression to metastasis, we here investigated the role of the matricellular protein transforming growth factor beta induced (TGFBI) in breast cancer. We crossed the MMTV-PyMT model of mammary gland tumorigenesis with a TgfbiΔ/Δ mouse and studied the CSC content of the tumours. We performed RNAseq on wt and ko tumours, and analysed the tumour vasculature and the immune compartment by IHC and FACS. The source of TGFBI expression was determined by qPCR and by bone marrow transplantation experiments. Finally, we performed insilico analyses using the METABRIC cohort to assess the potential prognostic value of TGFBI. We observed that deletion of Tgfbi led to a dramatic decrease in CSC content and lung metastasis. Our results show that lack of TGFBI resulted in tumour vessel normalisation, with improved vessel perfusion and decreased hypoxia, a major factor controlling CSCs and metastasis. Furthermore, human data mining in a cohort of breast cancer patients showed that higher expression of TGFBI correlates with poor prognosis and is associated with the more aggressive subtypes of breast cancer. Overall, these data reveal a novel biological mechanism controlling metastasis that could potentially be exploited to improve the efficacy and delivery of chemotherapeutic agents in breast cancer.

Comparison of the cardiac effects of electronic cigarette aerosol exposure with waterpipe and combustible cigarette smoke exposure in rats

AimsElectronic cigarette (ECIG) has been used as an alternative to tobacco smoking as it lacks the majority of toxicants found in tobacco smoke. However, the effect of ECIG aerosol inhalation on cardiac health are not well studied. The present study aimed to compare the effects of ECIGs with that of combustible tobacco cigarette (T-Cigs) and waterpipe (WP) smoke on cardiac biomarkers of oxidative stress, inflammation, and fibrosis. Main methodsRats were randomized into control (fresh air, n = 12), ECIG aerosol (n = 12), T-Cig smoke (n = 15), or WP (n = 13) smoke conditions in which they were exposed 1 h/daily, 6 day/week for 4 weeks. Cardiac biomarkers of oxidative stress, inflammation, and remodeling were assessed. Key findingsRelative to control, significant increase in heart to body weight ratio was observed in all exposed groups. Cardiac endothelin-1 and myeloperoxidase were increased for ECIG and T-Cig. Cardiac nitrite and TBARS were increased in all exposed groups, but activity of superoxide dismutase was increased for ECIG and T-Cig only while glutathione levels increased for ECIG only. No changes were observed for cardiac C-reactive protein and catalase activity. Cardiac fibrosis was observed in all exposed groups coupled with an increase in the transforming growth factor beta protein that was significant for ECIG only. SignificanceECIG aerosol may promote cardiac alterations in similar manner to tobacco smoke by promoting myocardial oxidative stress and inflammation leading to fibrosis. With regard to cardiac health, exposure to ECIG aerosol and combustible T-Cig smoke may lead to similar adverse outcomes.

Recombinant human PRG4 (rhPRG4) suppresses breast cancer cell invasion by inhibiting TGFβ-Hyaluronan-CD44 signalling pathway.

Metastasis is the major cause of cancer-related morbidity and mortality. The ability of cancer cells to become invasive and migratory contribute significantly to metastatic growth, which necessitates the identification of novel anti-migratory and anti-invasive therapeutic approaches. Proteoglycan 4 (PRG4), a mucin-like glycoprotein, contributes to joint synovial homeostasis through its friction-reducing and anti-adhesive properties. Adhesion to surrounding extracellular matrix (ECM) components is critical for cancer cells to invade the ECM and eventually become metastatic, raising the question whether PRG4 has an anti-invasive effect on cancer cells. Here, we report that a full-length recombinant human PRG4 (rhPRG4) suppresses the ability of the secreted protein transforming growth factor beta (TGFβ) to induce phenotypic disruption of three-dimensional human breast cancer cell-derived organoids by reducing ligand-induced cell invasion. In mechanistic studies, we find that rhPRG4 suppresses TGFβ-induced invasiveness of cancer cells by inhibiting the downstream hyaluronan (HA)-cell surface cluster of differentiation 44 (CD44) signalling axis. Furthermore, we find that rhPRG4 represses TGFβ-dependent increase in the protein abundance of CD44 and of the enzyme HAS2, which is involved in HA biosynthesis. It is widely accepted that TGFβ has both tumor suppressing and tumor promoting roles in cancer. The novel finding that rhPRG4 opposes HAS2 and CD44 induction by TGFβ has implications for downregulating the tumor promoting roles, while maintaining the tumor suppressive aspects of TGFβ actions. Finally, these findings point to rhPRG4’s potential clinical utility as a therapeutic treatment for invasive and metastatic breast cancer.

The SUMO System and TGFβ Signaling Interplay in Regulation of Epithelial-Mesenchymal Transition: Implications for Cancer Progression.

Protein post-translational modification by the small ubiquitin-like modifier (SUMO), or SUMOylation, can regulate the stability, subcellular localization or interactome of a protein substrate with key consequences for cellular processes including the Epithelial-Mesenchymal Transition (EMT). The secreted protein Transforming Growth Factor beta (TGFβ) is a potent inducer of EMT in development and homeostasis. Importantly, the ability of TGFβ to induce EMT has been implicated in promoting cancer invasion and metastasis, resistance to chemo/radio therapy, and maintenance of cancer stem cells. Interestingly, TGFβ-induced EMT and the SUMO system intersect with important implications for cancer formation and progression, and novel therapeutics identification.

Berberine Suppresses Cell Motility Through Downregulation of TGF-β1 in Triple Negative Breast Cancer Cells

Background/Aims: Transforming growth factor-beta proteins (TGF-βs) are multifunctional growth factors and powerful modulators of the epithelial-mesenchymal transition (EMT) in a variety of cancer types including breast and lung cancer cells. Here, we demonstrated the inhibitory effect of berberine (BBR) on tumor growth and metastasis of triple negative breast cancer (TNBC) cells via suppression of TGF-β1 expression. Methods: The levels of mRNA expression were analyzed by real-time PCR. The levels of MMP-2, MMP-9 and TGF-β1 protein expression were analyzed by zymography and confocal microscopy, respectively. Cell migration was analyzed by wound healing assay. Tumorigenicity of TNBC cells such as tumor growth and metastasis was analyzed using xenograft models. Results: In a clinical data set, aberrant TGF-β1 expression was associated with poor prognosis of breast cancer patients. Our in vitro results using TNBC cells showed that the expression levels of matrix metalloproteinase (MMP)-2 and MMP-9 and the capacity for cell migration were increased by TGF-β1 treatment. In contrast, basal levels of MMP-2 and MMP-9 were suppressed by a specific TGF-β receptor I inhibitor, SB431542. In addition, TGF-β1–induced MMP-2 and MMP-9 expression and cell migration were decreased by SB431542. Interestingly, we showed for the first time that BBR decreased the level of TGF-β1, but not TGF-β2, in TNBC cells. Furthermore, BBR significantly decreased the level of MMP-2 expression as well as the capacity for cell migration in TNBC cells. Finally, we examined the effect of BBR on in vivo tumor growth and lung metastasis in MDA-MB231 and 4T1 breast cancer xenograft models and showed that both were significantly decreased following BBR treatment. Conclusion: BBR suppresses tumorigenicity of TNBC cells through inhibition of TGF-β1 expression. Therefore, we demonstrate that BBR could be a promising drug for treatment of TNBC.

Collagen affinity coating for surface binding of decorin and other biomolecules: Surface characterization.

The development of biomaterials that promote tissue reconstruction and regeneration can reduce the low level, chronic inflammation and encapsulation that impact the performance of today's medical devices. Specifically, in the case of implantable sensors, the host response often leads to poor device performance that discourages permanent implantation. Our goal is to present on medical implants bioactive molecules that can promote healing rather than scarring. Localized delivery of these molecules would also minimize the possibility of adverse tissue reactions elsewhere in the body. Toward this end, the authors have developed a collagen affinity coating that binds a number of potential healing molecules and can be attached to the surface of an implanted biomaterial. This allows the creation of a wide variety of natural surface coatings that can be evaluated and tailored to promote the desired healing response. To demonstrate the efficacy of this collagen affinity coating to biospecifically bind promising healing molecules to type I collagen in vivo, the antifibrotic proteoglycan decorin was utilized. Decorin binds and renders ineffective the protein transforming growth factor beta (TGFβ) that induces collagen scar production. Thus, an assembled, supramolecular structure of biomaterial-collagen-decorin-TGFβ is formed. A decorin surface coating was created and characterized, illustrating the potential of this type I collagen affinity coating for widespread use with a variety of promising healing molecules. Future studies will test the implant efficacy of this type of coating.

Clinical significance of high levels of survivin and transforming growth factor beta-1 proteins in aqueous humor and serum of retinoblastoma patients

To evaluate the diagnostic and prognostic values of survivin and transforming growth factor beta-1 (TGF-B1) expression in aqueous humor and serum of retinoblastoma (RB) in comparison to the conventional RB marker lactate dehydrogenase (LDH) and to elucidate a possible correlation between them and the clinicopathological features of the disease. This prospective, comparative study included 88 newly diagnosed children with RB and 80 age-matched controls with ophthalmic conditions other than tumors prepared for intraocular surgeries. Concentrations of survivin, TGF-B1, and LDH were measured in serum and aqueous humor before and 6months after completion of therapy. High serum and aqueous humor concentrations of the three proteins were detected in RB patients before treatment compared to the control group (P<0.01), with a significant reduction of serum concentrations after treatment (P<0.01). For the highest sensitivity and specificity, the optimal cutoff values of serum and aqueous survivin were 12.9 pg/ml and 25.2 pg/mg, with a significant positive correlation between aqueous survivin and RB staging and presence of optic nerve infiltration (r=0.43, P=0.04); the best cutoff values of serum and aqueous TGF-B1, 370.7 pg/ml and 39.8 pg/mg, with a significant positive correlation between aqueous TGF-B1 and poor differentiation of the tumor (r=0.69, P=0.001). The high sensitivity, specificity, and accuracy of serum and aqueous humor survivin and TGF-B1 proteins make them promising markers for early detection and follow-up of RB patients.

Prophylactic effects of Clausena excavata Burum. f. leaf extract in ethanol-induced gastric ulcers.

Clausena excavata is a natural herb with both antioxidant and anti-inflammatory properties. It has been used for decades in folkloric practice for the amelioration of various ailments. In this study, the gastroprotective activity of methanolic extract of C. excavata leaves (MECE) was determined in the Sprague Dawley rat ethanol-induced gastric ulcer model. Rats were pretreated with a single dose of vehicle (5% Tween 20), 20 mg/mL omeprazole, 400 and 200 mg/mL of MECE dissolved in 5% Tween 20. Ulcer was induced with 5 mL/kg of ethanol and stomach tissue was obtained after 1 hour. Histological examination was done on hematoxylin and eosin, periodic acid-Schiff, and immunochemically stained gastric mucosal tissues. Prostaglandin E2, superoxide dismutase, catalase, glutathione peroxidase, and lipid peroxidation levels of the gastric tissue homogenates were also determined. Significantly (P<0.05) smaller ulcer areas, less intense edema, and fewer leukocytes’ infiltration were observed in MECE- and omeprazole-treated than in untreated gastric mucosa with ulcer. The gastric pH, mucus production, superoxide dismutase, catalase, and glutathione peroxidase contents increased, while the lipid peroxidation content decreased as a result of MECE treatment. Bcl-2-associated X protein was underexpressed, while heat shock protein 70 and transforming growth factor-beta protein were overexpressed in the ulcerated gastric mucosa tissues treated with omeprazole and MECE. Similarly, there was a reduction in the levels of tumor necrotic factor-alpha and interleukin-6, while the level of interleukin-10 was increased. This study showed that the gastroprotective effect of MECE is achieved through inhibition of gastric juice secretion and ulcer lesion development, stimulation of mucus secretion, elevation of gastric pH, reduction of reactive oxygen species production, inhibition of apoptosis in the gastric mucosa, and modulation of inflammatory cytokines.

Oral Presentation VIII

ABSTRACT BACKGROUND AND AIMS Activin, a member of the transforming growth factor beta protein family, was originally isolated from gonadal fluids and stimulates the release of pituitary follicle-stimulating hormone (FSH). Activin has numerous functions in both normal and neoplastic cells. Although it was suggested that gonadal tissue is the primary site of activin production, several extragonadal sources have subsequently been identified, including human thyrocytes. METHODS In our study, serum levels of activin A were measured by chemiluminescence, and the expression of receptors of activin A in thyroid tissue was detected by immunohistochemistry in female patients with thyroid papillary cancer [stage I (n = 60), stage II (n = 60), stage III (n = 60), stage IV (n = 60)] and in normal controls (n = 60). RESULTS The serum levels of activin A were no significantly different between patients with thyroid papillary cancer and normal controls as well as different stages. But the expressions of receptors of activin A were greater in patients with thyroid papillary cancer than in normal controls. And the positive rates of receptor expression were significantly more in stage III and IV than in stage I and II. CONCLUSION In conclusion, this study demonstrates that serum levels of activin A undergo no significant changes when thyroid papillary cancer occurs. The thyroid gland is not the predominant source of activin A. The expressions of receptors of activin A were significantly greater in patients with thyroid papillary cancer than in normal controls. And the positive rates of receptor expressions were associated with the severity of cancer. Because activin A may exert negative action on thyrocyte proliferation, it is conceivable that the increase in the receptor of activin A in thyroid papillary cancer might represent a counteracting mechanism.

Decorin gene expression in the differentiation of the skeletal connective tissues of the developing limb

Tendons and cartilages are connective tissues of essential importance in the musculoskeletal system. During digit development, cartilage and tendon primordia develop from the undifferentiated mesenchymal cells originated in the lateral plate mesoderm. The specification of these tissues begins with the establishment of cellular aggregates, which prefigure the tendons and phalanges. Transforming growth factor beta proteins (TGFβs) are the inductive signals responsible not only for the initiation of chondrogenesis and tenogenesis during digit formation, but also for joint specification. An early role of this family of secreted proteins during these processes is to promote mesenchymal cell precursors condensation. Here we show that Decorin presents an overlapping pattern of expression with TGFβ2 in joint and tendon blastemas of the embryonic digits. Furthermore, Decorin expression is induced by TGFβ signaling, and DECORIN promotes aggregation of digit mesenchymal cell precursors. In addition, we provide gene expression studies suggesting that Cadherin-11 may function as an effector of Decorin in this experimental model.

Prognostic value of tgfb1 protein in endometrioid adenocarcinoma

Angiogenesis is a prerequisite for tumour development, progression and metastasis; however, its underlying molecular mechanisms in endometrial carcinoma are poorly understood. In this study, the mRNA and protein expression profiles of two key regulators of angiogenesis, vascular endothelial growth factor (VEGF) and transforming growth factor beta-1 (TGFB1), were evaluated by real-time PCR and western blot analysis in 23 endometrial cancer tissue-paired specimens (malignant vs. adjacent normal tissues). We aimed to investigate whether VEGF and TGFB1 serve as markers of the malignant transformation of the endometrium and whether VEGF or TGFB1 expression can constitute a useful prognostic marker of survival in patients with endometrial carcinoma. Tissue-pair analysis revealed VEGF transcriptional up-regulation and TGFB1 mRNA down-regulation as the most frequent transcriptional features. VEGF and TGFB1 mRNA were positively correlated (P < 0·001). VEGF protein levels were higher in endometrioid-type tissue pairs (P = 0·047). TGFB1 protein and mRNA levels were negatively correlated (P = 0·042). TGFB1 protein expression was related to survival only in patients with endometrioid adenocarcinoma (P = 0·045). Tissue-pair mRNA and protein analysis reveals VEGF transcriptional up-regulation and TGFB1 down-regulation that are correlated with the malignant transformation of the endometrium, while post-transcriptional mechanisms control VEGF and TGFB1 protein. TGFB1 protein demonstrated a prognostic value only in endometrioid adenocarcinoma.