Expression of transforming growth factor alpha in human tissues: Immunohistochemical study and Northern blot analysis

Summary The expression of transforming growth factor alpha (TGF-) was examined in various human tissues and the fetus, using immunohistochemistry and Northern blot analysis. TGF- immunoreactivity was detected mainly in the epithelial cells of the digestive tract, liver, pancreas, kidney, thyroid, adrenal, skin, mammary gland and genital organs. In the digestive tract, epithelial cells with regenerative change or hyperplastic change showed strong immunoreactivity to TGF-. Peripheral nerve, vessels, megakaryocytes and macrophages in the lung and spleen were also positive for TGF-. By Northern blot analysis the expression of TGF- mRNA was confirmed in the digestive tract, salivary gland, thyroid, kidney and mammary gland. In the human fetus, the nerve tissues, liver, adrenal and kidney were positive for TGF-. Strong immunoreactivity to TGF- was observed in the hepatocytes of the fetus. These findings indicate that TGF- is produced by a variety of nonneoplastic cells in both adult and fetal tissues.     

Immunohistochemical analysis of hepatocyte growth factor and c-Met in plasma cell disease

Wader K F, Fagerli U‐M, Børset M, Lydersen S, Hov H, Sundan A, Bofin A & Waage A
(2012) Histopathology 60 ,443–451
Immunohistochemical analysis of hepatocyte growth factor and c‐Met in plasma cell disease Aims: Interaction with the bone marrow microenvironment is important for homing and survival of myeloma cells. One cytokine involved in this process is hepatocyte growth factor (HGF). HGF, by binding to the receptor tyrosine kinase c‐Met, mediates a broad range of tumour progression activities. Our aims were to investigate whether HGF and c‐Met are present in bone marrow and extramedullary tumours from patients with monoclonal plasma cell disease, and whether c‐Met is activated. Methods and results: Expression of HGF, c‐Met and phospho‐c‐Met was studied by immunohistochemistry in biopsies from 80 patients with monoclonal plasma cell disease. Cytoplasmic staining for HGF in plasma cells was demonstrated in 58 of 68 biopsies from multiple myeloma patients (85%), but also in biopsies from nine of 10 healthy individuals. Membranous staining for c‐Met was found in 25 of 63 multiple myeloma patients (40%) and in none of 10 healthy individuals. Membranous staining for phospho‐c‐Met was found in biopsies from 15 of 21 c‐Met‐positive myeloma patients (71%). Conclusions: Our data point to c‐Met expression as one of the factors that distinguishes normal from malignant plasma cells, and indicate that the HGF/c‐Met system is activated in multiple myeloma patients.     

转化生长因子β1对乳鼠心肌细胞转化生长因子结合蛋白2表达的影响

 目的：探讨转化生长因子β1（TGF-β1）在诱导心肌细胞表达转化生长因子结合蛋白2（LTBP2）中的作用及信号传导通路。 方法：培养乳鼠心肌细胞;实时定量聚合酶链式反应（Real-time PCR）、蛋白质印迹和免疫细胞化学方法检测不同时间和不同浓度的TGF-β1对大鼠乳鼠心肌细胞LTBP2基因及蛋白表达的影响;用TGF-β1相关信号通路阻断剂探讨TGF-β1调节LTBP2表达改变的信号传导机制。 结果：LTBP2基因表达随着TGF-β1浓度增加（0、2、5、10 ng/mL）而明显升高,在5 ng/mL时刺激最强（P < 0.05）;5 ng/mL的TGF-β1刺激下心肌细胞内LTBP2基因和蛋白表达的升高呈时间依赖性,均在12 h最高,24 h开始呈下降趋势（P < 0.05或P<0.01）;免疫细胞化学结果显示TGF-β1明显升高LTBP2的表达。信号传导通路研究显示TGF-β1在心肌细胞内主要通过ERK信号通路和PI3K信号通路诱导LTBP2的表达。 结论：TGF-β1在乳鼠心肌细胞内通过ERK信号通路和PI3K信号通路上调LTBP2的表达。     

Two novel polymorphisms in the human transforming growth factor beta 2 gene

We have identified two novel polymorphisms in the transforming growth factor beta 2 (TGFbeta2) gene; an insertion in the 5'-untranslated region (5'UTR) and a single nucleotide polymorphism (SNP) in exon 1. A 895-bp fragment was analysed covering part of the 5'UTR and exon 1. Single-strand conformation polymorphism (SSCP) analysis of polymerase chain reaction (PCR) products was performed to detect sequence variations. This was followed by the sequencing of samples demonstrating distinct banding patterns. A 4-bp insertion (ACAA) in the 5'UTR and a SNP (G > A) within exon 1 was identified. The 5'UTR polymorphism was found to be common in three Caucasian populations from Spain, Turkey and the UK. Exon 1 polymorphism is rare and results in an R to H amino acid substitution in codon 91. Both polymorphisms may prove useful for investigating possible associations of TGFbeta2 with disease.     

Effect of transforming growth factor beta 1 on neonatal rat latent transforming growth factor beta binding protein 2 in cardiomyocytes

目的 探讨转化生长因子β1 (TGF-β1)在诱导心肌细胞表达转化生长因子结合蛋白2(LTBP2)中的作用及信号传导通路.方法 培养乳鼠心肌细胞;实时定量聚合酶链式反应、蛋白质印迹和免疫细胞化学方法检测不同时间和不同浓度的TGF-β1对大鼠乳鼠心肌细胞LTBP2基因及蛋白表达的影响;用TGF-β1相关信号通路阻断剂探讨TGF-β1调节LTBP2表达改变的信号传导机制.结果 LTBP2基因表达随着TGF-β1浓度增加(0、2、5及10 μg/L)而明显升高,在5μg/L时刺激最强(P＜0.05);5μg/L的TGF-β1刺激下心肌细胞内LTBP2基因和蛋白表达的升高呈时间依赖性,均在12 h最高,24 h开始呈下降趋势(P＜0.05或P＜0.01);免疫细胞化学结果显示TGF-β1明显升高LTBP2的表达.信号传导通路研究显示TGF-β1在心肌细胞内主要通过ERK信号通路和PI3K信号通路诱导LTBP2的表达.结论TGF-β1在乳鼠心肌细胞内通过ERK信号通路和PI3K信号通路上调LTBP2的表达.     

Salivary gland expression of transforming growth factor beta isoforms in Sjogren's syndrome and benign lymphoepithelial lesions.

AIM: Transforming growth factor beta (TGF-beta) is involved in the control of autoimmune reactions, cell proliferation, and the accumulation of lymphocytes within organs. The aim of this study was to determine the expression of TGF-beta in salivary glands from patients with primary Sjogren's syndrome (SS) and benign lymphoepithelial lesions (BLEL) with emphasis on ductal epithelium. METHODS: Immunoperoxidase staining for TGF-beta isoforms and Ki67 antigen was performed on formalin fixed sections of labial glands from patients with primary SS (n = 15) and controls (n = 5) and parotid glands reported as BLEL (n = 5) or normal (n = 5). Ductal expression of TGF-beta was quantified by absorbance measurements using image analysis. The specificity of staining was confirmed by peptide blocking studies. RESULTS: All TGF-beta isoforms were detected within the cytoplasm of most lymphocytes, endothelial cells, and ducts in all specimens. Acinar expression was variable and weaker than that seen in ducts. Absorbance measurements revealed that the expression of all isoforms was greater in ducts within primary SS glands than in control glands. Ductal expression in control parotid glands was greater than that seen in BLEL glands, irrespective of the presence of adjacent lymphoid infiltrates. Comparisons between control specimens showed that ductal expression of all isoforms was highest in parotid glands, whereas no differences were detected between primary SS and BLEL glands. Ki67 positive lymphocytes and duct cells were mainly restricted to pathological specimens, with BLEL glands containing larger populations of positive cells than primary SS glands. CONCLUSION: These results demonstrate complex and variable changes in ductal expression of TGF-beta in primary SS and BLEL, which may be important in the control of lymphoid infiltration and the proliferation of lymphocytes and ductal epithelium.     

Fibroblast growth factor 2 and transforming growth factor beta1 synergism in human bronchial smooth muscle cell proliferation

Bronchial smooth muscle cell (BSMC) hyperplasia is a typical feature of airway remodeling and contributes to airway obstruction and hyperresponsiveness in asthma. Fibroblast growth factor 2 (FGF-2) and transforming growth factor beta1 (TGF-beta1) are sequentially upregulated in asthmatic airways after allergic challenge. Whereas FGF-2 induces BSMC proliferation, the mitogenic effect of TGF-beta1 remains controversial, and the effect of sequential FGF-2 and TGF-beta1 co-stimulation on BSMC proliferation is unknown. This study aimed to assess the individual and sequential cooperative effects of FGF-2 and TGF-beta1 on human BSMC proliferation and define the underlying mechanisms. Mitogenic response was measured using crystal violet staining and [3H]-thymidine incorporation. Steady-state mRNA and protein levels were measured by semiquantitative RT-PCR, Western blot, and ELISA, respectively. TGF-beta1 (0.1-20 ng/ml) alone had no effect on BSMC proliferation, but increased the proliferative effect of FGF-2 (2 ng/ml) in a concentration-dependent manner (up to 6-fold). Two distinct platelet-derived growth factor receptor (PDGFR) inhibitors, AG1296 and Inhibitor III, as well as a neutralizing Ab against PDGFRalpha, partially blocked the synergism between these two growth factors. In this regard, TGF-beta1 increased PDGF-A and PDGF-C mRNA expression as well as PDGF-AA protein expression. Moreover, FGF-2 pretreatment increased the mRNA and protein expression of PDGFRalpha and the proliferative effect of exogenous PDGF-AA (140%). Our data suggest that FGF-2 and TGF-beta1 synergize in BSMC proliferation and that this synergism is partially mediated by a PDGF loop, where FGF-2 and TGF-beta1 upregulate the receptor (PDGFRalpha) and the ligands (PDGF-AA and PDGF-CC), respectively. This powerful synergistic effect may thus contribute to the hyperplastic phenotype of BSMC in remodeled asthmatic airways.     

TGF-β1及TGF βR Ⅱ mRNA在子痫前期患者胎盘组织中的表达及意义

目的检测胎盘组织中TGF-β1(transform ing growth factor-beta1,TGF-β1)及其受体TGFβRⅡ(TGF-beta receptorⅡ)的表达,探求TGF-β1在子痫前期发病中的作用。方法用半定量逆转录聚合酶链反应(RT-PCR)技术对10例正常妊娠组和26例子痫前期组(包括12例轻度和14例重度)胎盘组织中TGF-β1、TGFβRⅡmRNA的表达水平进行检测,并通过紫外凝胶图像分析进行定量分析。结果重度子痫前期组TGF-β1和TGFβRⅡmRNA的表达量均升高,与轻度子痫前期组及正常妊娠组相比有明显意义(P<0.05)。结论TGF-β1、TGFβRⅡ在转录水平就已经上调,它们的异常表达它们可能与子痫前期的发病有着重大的关系。     

Immunohistochemical study of transforming growth factor beta, fibronectin, and fibronectin receptor in invasive mammary carcinomas.

Thirty-one cases of mammary carcinoma were examined immunohistochemically for the expression of transforming growth factor (TGF) beta, fibronectin (FN) and fibronectin receptor (FNR) in order to clarify the reason for the reported relationship between TGF beta expression and a high incidence of lymph node metastasis. It was revealed that TGF beta expression is closely related to the expression of FN, an intercellular matrix protein, and its cellular receptor FNR, one of the integrins. The interaction between FN and FNR in a tumor is considered to form the basis of the invasive nature of carcinoma cells. Thus, it is suggested that TGF beta expression in carcinoma cells induces the interaction between FN and FNR, which may lead to carcinomatous invasion resulting in lymph nodal metastasis.     

软骨形成过程中的转化生长因子β2

背景：转化生长因子β2是一族多肽类生长因子,具有多重生物学效应,对骨髓间充质干细胞增殖分化功能有一定促进作用,是调节骨髓间充质干细胞增殖和向软骨细胞方向分化的最主要因子之一。 目的：探讨转化生长因子β2 的分子结构特点及其在软骨形成方面的作用。 方法：应用计算机检索中国期刊全文数据CNKI、中国期刊全文数据维普中文科技期刊数据库(VIP)和PubMed数据库(1995-01/2011-08)与骨组织工程中转化生长因子β2 诱导软骨形成有关的文章,检索词分别为“转化生长因子β2;软骨细胞分化;骨组织工程”和“TGF-β2;cartilage formation;tissue engineering”。纳入所述内容与转化生长因子β2 分子结构特点,转化生长因子β2信号转导通路,软骨细胞的诱导性分化,软骨性疾病的生物性治疗进展有关。排除综述文献、重复研究、观点陈旧的文章。 结果与结论：初检得到302篇文献,排除262篇不符合标准的文献,共纳入40篇符合标准的文献。经分析得出转化生长因子β2通过促进骨髓间充质干细胞分化为软骨细胞促进软骨特异性基质如Ⅱ型胶原及蛋白多糖的合成,从而发挥软骨诱导作用。转化生长因子β2 与其他因子共同作用调节软骨细胞生长分化,使临床上永久性修复软骨组织缺损的治疗变为可能。     

Regulation and regulatory activities of transforming growth factor beta

Transforming growth factor beta (TGF beta) is unique among growth factors in its potent and widespread actions. Almost every cell in the body has been shown to make some form of TGF beta, and almost every cell has receptors for TGF beta. Therefore, it becomes apparent that this growth factor must be tightly regulated to prevent disease. The mechanisms of regulation of TGF beta are extensive and complex. One set of mechanisms centers around the fact that TGF beta is produced in a latent form that must be activated to produce biologically active TGF beta. These mechanisms include the latency of the molecule, the production of various latent forms, its targeting to cells for activation or to matrix for storage, and the means of activation of the latent forms. The TGF beta isoforms and the types, affinity, and signaling functions of its receptors also add complexity to the regulation of the effects of TGF beta. Active TGF beta regulates numerous processes in the body. TGF beta has three major biological effects: growth inhibition, stimulation of extracellular matrix formation, and immunosuppression. The means by which TGF beta regulates the expression of the numerous genes on which it has effects are complex and more information is needed. The means by which TGF beta regulates gene expression and the means by which the actions of TGF beta are regulated are addressed in this review.     

Immunosuppressive role of transforming growth factor beta in breast cancer

Transforming growth factor beta (TGF-β) is a multifunctional cytokine, whose myriad of functions include its ability to potently suppress the immune system. Because of its ability to negatively modulate the inductive and effector phases of the immune response, TGF-β is thought to contribute to tumor progression and metastases formation. Immunosuppression by tumor-derived TGF-β is increasingly becoming recognized as an important factor in tumor progression and may, in part, explain the low response rates achieved in cancer patients undergoing immunotherapy for their disease. This review will focus on the immunosuppressive role of tumor-derived TGF-β in breast cancer. Due to the paucity of human studies, it will specifically address the actions of tumor-derived TGF-β on cells of the immune system in preclinical animal models, as well as discuss strategies to negate the deleterious effects of TGF-β in order to improve the anti-tumor immune response.     

Immunohistochemical evidence of a role for transforming growth factor beta in the pathogenesis of nodular sclerosing Hodgkin's disease.

Transforming growth factor beta (TFG-beta) is a multifunctional growth factor that promotes the growth of fibroblasts, collagen synthesis and angiogenesis, and stimulates monocyte migration and activation, but suppresses the growth and differentiation of immune lymphocytes and killer cells. Previously we demonstrated biologic activity for TGF-beta in supernatants of fresh Hodgkin's disease (HD) cell cultures and the cell line L428 derived from nodular sclerosing HD. This study was undertaken to find evidence of TGF-beta activity directly in tissues affected by HD. Formalin-fixed tissue from 14 patients with HD, including 8 nodular sclerosis, 4 mixed cellularity, 1 lymphocyte predominance, and 1 lymphocyte depletion type were studied by immunoperoxidase technique with antibody CC (1-30) raised against a synthetic polypeptide with the same N-terminal amino acid sequence as TGF-beta 1. Transforming growth factor beta activity was demonstrated in six cases of nodular sclerosis but not in other histologic types of HD. Staining for TGF-beta was found in the cytoplasm of Reed-Sternberg (RS) cells in one case and on the surface of RS cells and their lacunar variants in five cases. Transforming growth factor beta activity associated with the extracellular matrix was localized mainly around blood vessels, zones of necrosis, at the margins of bands of collagen sclerosis, and in areas containing syncytia of RS cells. In two cases TGF-beta was associated with collections of epithelioid histiocytes or granulomas. Small lymphocytes, granulocytes, and germinal center cells were unreactive. These results suggest that TGF-beta is a growth factor of biologic importance in HD and may be responsible for many of the histologic features, such as nodular sclerosis and granulomas, that may have prognostic significance.     

Bioactivation of latent transforming growth factor beta1 by Mycobacterium tuberculosis in human mononuclear phagocytes

Biologically active transforming growth factor beta 1 (TGFbeta1) has been identified at sites of Mycobacterium tuberculosis (MTB) infection in the lung; however, the underlying mechanism(s) for its activation is not clear. Here using an enzyme-linked immunospot assay for TGFbeta1, we show that human blood monocytes (MN) and alveolar macrophages (AM) produce bioactive TGFbeta1 upon stimulation by MTB. However, only MTB-stimulated MN increased TGFbeta1 production on a per cell basis. The frequency of TGFbeta1-producing MN was reduced by an inhibitor of plasmin, bdellin, indicating a role for plasmin pathways in the bioactivation of cytokine. The expression of urokinase plasminogen activator receptor (uPAR) mRNA and both surface and soluble uPAR (CD87) was increased in MTB-activated MN. However, antibody neutralization of uPAR suppressed bioactive TGFbeta1 in MN alone. Thus, the more immature MN, which are continuously recruited to the lung during tuberculosis (TB), have a higher capacity to bioactivate TGFbeta1 by expression of components of the plasmin pathway. Excess production and bioactivation of TGFbeta1 at sites of MTB infection may undermine host immune responses during TB.     

Immunohistochemical expression of transforming growth factor alpha and epidermal growth factor receptor in pancreatic endocrine tumors.

Coexpression of transforming growth factor alpha (TGF-alpha) and its receptor epidermal growth factor receptor (EGFR) is known to be associated with aggressive biologic behavior and adverse clinical outcome in a variety of tumors, including pancreatic adenocarcinomas. However, very little information is currently available as to whether this is true of pancreatic endocrine tumors (PETs) as well. Thirty-five PETs were retrospectively studied for immunohistochemical expression of TGF-alpha, the intracellular and extracellular domains of EGFR, and various hormonal secretory products. Proliferative activity was additionally studied (in 20 cases only) using the MIB-1 antibody. Thirty-one (89%) of 35 tumors were reactive for 1 or more of the peptide hormones tested; 22 (63%) tumors were positive for TGF-alpha; and 23 (65%) were positive for the intracellular and/or extracellular domain of EGFR. Based on their TGF-alpha and EGFR expression, these tumors could be classified into 4 groups. Of the 10 tumors in group I (positive for TGF-alpha and the complete EGFR molecule), 3 were malignant, 6 were >2 cm in diameter, 5 were functional, and 1 had a proliferative index of >40%. The 12 tumors in group II (positive for TGF-alpha but negative for the intracellular and/or extracellular domain of EGFR) included 4 malignant tumors, 4 PETs >2 cm in diameter, 8 functional, and 1 with a proliferative index of >40%. The 7 PETs in group III (positive for the intracellular/extracellular domain of EGFR alone) included 3 malignant tumors, 3 PETs >2 cm in diameter, and 3 functional tumors. The 6 tumors in group IV (completely negative for both TGF-alpha and EGFR) included 4 malignant tumors, 3 PETs >2 cm in diameter, and 4 functional lesions. Therefore, immunohistochemical expression of TGF-alpha and EGFR, either alone or in concert, shows no correlation with size, functional status, secretory profile, or biologic behavior and hence cannot be used as a marker of malignancy in this group of tumors.     

转化生长因子β1在兔房水中的浓度变化

背景：转化生长因子β1可参与角膜损伤后的修复。 目的：观察转化生长因子β1滴眼液滴眼后房水中的浓度变化规律。 方法：将新西兰大白兔随机分为5组,分别给予PBS和质量浓度为0.5,1.0,2.0,4.0 mg/L的转化生长因子β1滴眼液滴右眼。 结果与结论：通过裂隙灯观察兔角膜和结膜结构,各组兔眼均无结膜分泌物、球结膜充血、角膜水肿增厚、角膜后沉着物、前房炎性反应及晶状体混浊改变。ELISA检测结果显示,与PBS组比较,质量浓度2.0和4.0 mg/L转化生长因子β1滴眼液能有效提高兔眼房水中转化生长因子β1的质量浓度(P < 0.01),角膜穿透性良好,在房水中可以达到有效的治疗浓度。