Growth-promoting Factors Research Articles

Evidence for a role of sphingosine-1 phosphate in cardiovascular remodelling in Fabry disease

A hallmark of Fabry disease is the concomitant development of left-ventricular hypertrophy and arterial intima-media thickening, the pathogenesis of which is thought to be related to the presence of a plasmatic circulating growth-promoting factor. We therefore characterized the plasma of patients with Fabry disease in order to identify this factor. Using a classical biochemical strategy, we isolated and identified sphingosine-1 phosphate (S1P) as a proliferative factor present in the plasma of patients with Fabry disease. Plasma S1P levels were significantly higher in 17 patients with Fabry disease compared with 17 healthy controls (225 +/- 40 vs. 164 +/- 17 ng/mL; P = 0.005). There was a positive correlation between plasma S1P levels and both common carotid artery intima-media thickness and left-ventricular mass index (r(2) = 0.47; P = 0.006 and r(2) = 0.53; P = 0.0007, respectively). In an experimental model, mice treated with S1P developed cardiovascular remodelling similar to that observed in patients with Fabry disease. Sphingosine-1 phosphate participates in cardiovascular remodelling in Fabry disease. Our findings have implications for the treatment of cardiovascular involvement in Fabry disease.

Genetic diversity and characterization of heavy metal-resistant-endophytic bacteria from two copper-tolerant plant species on copper mine wasteland

The diversity of endophytic bacteria from the copper-tolerant species Elsholtzia splendens and Commelina communis was evaluated by using cultivation and cultivation-independent techniques. Phylogenetic analysis based on 16S rDNA sequences showed that the Cu-resistant isolates belonged to three major phylogenetic groups: Firmicutes, Actinobacteria and Proteobacteria. Bacillus and Acinetobacter were predominant among the Cu-resistant isolates. Sequence analysis from the 16S rDNA clone libraries of the two plant roots revealed sequences of Alpha-, Beta-, Gamma- Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes. Gammaproteobacteria was predominant in the two 16S rDNA clone libraries. Thirty-two endophytic bacteria were isolated and characterized with respect to heavy metal resistance and production of plant growth-promoting factors. In experiments involving rape plants grown in vermiculite containing 4 mg kg −1 of Cu, inoculation with the isolates was found to increase the dry weights of roots (ranging from 132% to 155%) and above-ground tissues (ranging from 71% to 83%) compared to the uninoculated control. Increase in above-ground tissue Cu content varied from 63% to 125% in inoculated-rape plants cultivated in Cu-contaminated substrate compared to the uninoculated control.

Effect of interleukin-1a, IL-1b and IL-1 receptor antibody on the proliferation and steroidogenesis of regenerating rat adrenal cortex.

Several data suggest that the endocrine and immune systems are closely interconnected, therefore, we investigated the effect of interleukin-1 on adrenocortical regeneration. The study was performed on adult rats subjected to left adrenal enucleation combined with contralateral adrenalectomy. The animals were given purified human recombinant IL-1 alpha (5 micrograms/kg), IL-1 beta (5 micrograms/kg) anti-human IL-1 receptor antibody (10 micrograms/kg) and combination of interleukins and antibodies. The cell proliferation measured by bromodeoxyuridine incorporation and plasma corticosterone determined by a RIA kit were evaluated six days after operation. The mean cell growth ratio expressed as number of BrdU-positive cells per 1,000 cells was 11.86 +/- 0.45 in the control group. IL-1b significantly stimulated the proliferation ratio (15.30 +/- 0.88; p < 0.01) and this effect could be prevented by addition of IL-1rAb. The antibody caused also a well-marked inhibition of cell growth ratio (8.23 +/- 0.67; p < 0.01). The plasma corticosterone concentration was higher in IL-1b-treated animals than in the control group, 107.79 +/- 2.82 vs. 61.08 +/- 2.0 ng/ml, respectively, p < 0.01. On the other hand, IL-1rAb decreased corticosterone secretion (31.53 +/- 2.58 ng/ml, p < 0.01) and reversed the stimulatory effect of of IL-1b. In contrast, IL-1a seemed to have no influence on adrenocortical regeneration. In conclusion, this study supported the thesis of close interconnection between immune and endocrine systems and revealed that IL-1b could act as a growth-promoting factor on regenerating adrenal cortex in the rat.

Expression of apoptosis-related proteins in thyroid tumors and thyroid carcinoma cell lines.

Whereas in normal human thyroid tissue total cell mass is maintained by a balance between cell proliferation and apoptosis, the programmed cell death, in thyroid tumors this equilibrium is disrupted. In tumor cells, an augmented proliferation rate is no longer counterbalanced by an equally enhanced apoptosis resulting in an increased netto growth rate. To investigate regulation of apoptosis in thyroid tumors, we analyzed the expression of apoptosis-related proteins of the bcl-2 family in human thyroid tissues and in the human thyroid carcinoma cell lines FTC 133, HTC, HTC-TSHr and HTh74. In comparison to normal tissue, we detected an increased expression of the anti-apoptotic protein bcl-2 in adenomas, whereas follicular carcinomas showed various expression of bcl-2 with decreased levels in 32% of cases. BclxL expression was comparable in all tissues examined. The pro-apoptotic protein bax was expressed at lower levels in carcinomas than in adenomas, whereas bak and bclx were expressed in the same order of magnitude in all tissues examined. In contrast, thyroid carcinoma cell lines exhibited a relatively strong expression of bclxL, but a weak expression of bcl-2. In all four cell lines, the amounts of the pro-apoptotic proteins bax, bak and bclx were higher than in most tumor tissues. Our data show that in thyroid tumors expression of members of the bcl-2 protein family is not uniform. Rather, the expression pattern of pro- and anti-apoptotic proteins in thyroid tumors is heterogeneous. This may, at least in part, reflect the futile attempt of tumor cells to counterbalance the action of growth-promoting factors in thyroid tumor-igenesis.

Illuminated behaviour: phytochrome as a key regulator of light foraging and plant anti‐herbivore defence

In many ecological scenarios, the success of an individual plant is defined by the behavioural decisions that it makes when confronted with the risks of competition with other plants, and biomass losses to insect herbivores. These decisions involve expression of shade avoidance responses and induced chemical defences. Because these responses are costly, they frequently engender resource allocation dilemmas. In this review, I discuss the mechanisms that trigger adaptive responses to competitors and herbivores, highlighting the role of phytochromes as central organizers of the overall resource allocation strategy of plants. Phytochromes sense the reduction in the red to far-red (R : FR) ratio of sunlight caused by the proximity of other plants. Shade-intolerant plants respond to low R : FR ratios with shade avoidance behaviours and reduced investment in defence. Pfr depletion leads to increased stability of growth-promoting phytochrome-interacting factors (PIFs), and results in the production of auxins and gibberellins, degradation of DELLA proteins, which are repressors of PIFs, and reduced sensitivity to jasmonates. Thus, phytochrome appears to fulfil its organizational role by regulating the relative strength of the signalling circuits controlled by growth-related and defence-related hormones. I point out cases of signalling redundancy and discuss the significance of recent work on hormone signalling for our understanding of the mechanisms that control adaptive plant behaviour.

YB-1 Translational Control of Epithelial-Mesenchyme Transition

Transitions between epithelial and mesenchmal phenotypes play critical roles in normal development and cancer progression. In this issue of Cancer Cell, Evdokimova et al. demonstrate that YB-1 regulates epithelial-mesenchyme transition (EMT) by inducing cap-independent translation of mRNAs encoding EMT-promoting factors and suppressing cap-dependent translation of mRNAs encoding growth-promoting factors.

Embryonic substantia nigra grafts in the mesencephalon send neurites to the host striatum in non‐human primate after overexpression of GDNF

In spite of partial success in treating Parkinson's disease by using ectopically placed grafts of dopamine-producing cells, restoration of the original neuroanatomical circuits, if possible, might work better. Previous evidence of normal anatomic projections from ventral mesencephalic (VM) grafts placed in the substantia nigra (SN) has been limited to neonatal rodents and double grafting or bridging procedures. This study attempted to determine whether injection of a potent growth-promoting factor, glial cell line-derived neurotrophic factor (GDNF), into the target regions or placement of fetal striatal co-grafts in the nigrostriatal pathway might elicit neuritic outgrowth to the caudate nucleus. Four adult St. Kitts green monkeys received embryonic VM grafts into the rostral mesencephalon near the host SN, and injections of adeno-associated virus 2 (AAV2)/GDNF or equine infectious anemia virus (EIAV)/GDNF into the caudate. Three adult monkeys were co-grafted with fetal VM tissue near the SN and fetal striatal grafts (STR) 2.5 mm rostral in the nigrostriatal pathway. Before sacrifice, the striatal target regions were injected with the retrograde tracer Fluoro-Gold (FG). FG label was found in tyrosine hydroxylase-labeled neurons in VM grafts in the SN of only those monkeys that received AAV2/GDNF vector injections into the ipsilateral striatum. All monkeys showed FG labeling in the host SN when FG labeling was injected on the same side. These data show that grafted dopaminergic neurons can extend neurites to a distant target releasing an elevated concentration of GDNF, and suggest that grafted neurons can be placed into appropriate loci for potential tract reconstruction.

Identification and characterization of human ARIP2 and its relation to breast cancer

Activin, a member of the TGF-β superfamily, inhibits the proliferation of breast cancer cells. Activin interacts with its type I and type II receptors to induce phosphorylation of intracellular signaling molecules known as Smads. Previous studies showed that mouse ARIP2 can reduce activin signaling by interacting with activin type II receptors (ActRIIs); however, the activity of ARIP2 in breast cancer is still unclear. In this study, we used RT-PCR to obtain a human homologue of mouse ARIP2, human activin receptor-interacting protein 2 (hARIP2). Like murine ARIP2, hARIP2 has a PDZ domain in its NH2-terminal region and can interact specifically with ActRIIs. Overexpression of hARIP2 reduced activin-induced transcriptional activity and enhanced cell proliferation and colony formation in human breast adenocarcinoma MCF-7 cells and MDA-MB-231 cells. However, down-regulation of hARIP2 expression by RNAi enhanced activin-induced transcriptional activity and reduced cell proliferation and colony formation. Immunohistochemistry revealed that hARIP2 was expressed more frequently and much more intensely in malignant breast tissues such as simple carcinoma, invasive ductal carcinoma and mucinous adenocarcinoma than in benign hyperplasia or fibroadenoma cases. These results suggest that hARIP2 is a putative growth-promoting factor involved in breast tumorigenesis and tumor development.

Diethylphthalate, Possible Interactions in Fetal Brain Development

Many natural and synthetic compounds, including a variety of the chemicals used as plasticizers or in the pro- duction of cosmetics and therapeutics, have steroid agonist or antagonist activities, altering hormone-regulated gene ex- pression. The phthalate (diethylphthalate, extensively used as a plasticizer and in consumer products, are evaluated using the human renal epithelial cell line 293T/17. Emphasis of the study was on genes essential for central nervous system de- velopment or function. Cells were treated with 1, 10 or 100 μM phthalate and gene expression was measured in treated cells, showing significant up- or down-regulation of a large number of genes in treated compared to untreated cells. Of the 19,000 human genes on the DNA array chip utilized, two specific genes, FGD1 and NGPF2, were selected to corroborate mRNA levels using quantitative real time PCR (qrtPCR) data to confirm results obtained from the microarray determina- tions. FGD1 (faciogenital dysplasia) and NGPF2 (neurite growth-promoting factor 2, also called Midkine, MDK), showed a significant, possibly estrogen-synergistic, down-regulation of genes essential for fetal brain development. These studies were designed to provide data on the gene expression-altering capacity of a widely distributed chemical, diethylphthalate (DEP), and to show possible associations between the previously reported widespread presence of DEP and the DEP me- tabolite, MEP, in urine samples from a reference population, the potential for altered gene expression in human cells in vi- tro, and possible neurodevelopmental effects that could be correlated with in utero exposure to DEP.

Exogenous CNTF stimulates axon regeneration of retinal ganglion cells partially via endogenous CNTF

Intravitreal injections of exogenous CNTF stimulate axon regeneration of RGCs in vivo. Nevertheless, controversy exists over the ability of exogenous CNTF to directly stimulate axon regeneration of mature RGCs. Here we demonstrate that CNTF potently stimulated axon outgrowth of mature RGCs in culture in a JAK/STAT3- and PI3K/AKT-signaling pathway-dependent fashion and stronger than oncomodulin. Additional cAMP elevation or inhibition of MAPK activity increased these effects. In vivo intravitreal injections of exogenous CNTF induced endogenous CNTF expression in astrocytes in a manner that depended on the MAPK/ERK-signaling pathway activation. Reduction of endogenous CNTF expression by MAPK/ERK pathway inhibitors or its absence in CNTF deficient mice markedly reduced the neurite growth-promoting effects of exogenous CNTF. These data demonstrate that CNTF is a potent axon growth-promoting factor for mature RGCs. However, exogenously applied CNTF stimulates RGCs in vivo partially indirectly via a mechanism that depends on astrocyte-derived CNTF.

Up-Regulation of Cyclooxygenase-2 Expression and Prostaglandin E2 Production in Human Endometriotic Cells by Macrophage Migration Inhibitory Factor: Involvement of Novel Kinase Signaling Pathways

Cyclooxygenase (COX) is the rate-limiting enzyme in the metabolic conversion of arachidonic acid to prostaglandins (PGs), including prostaglandin E(2) (PGE(2)), a major mediator of inflammation and angiogenesis. Herein, we report that macrophage migration inhibitory factor (MIF), a potent proinflammatory and growth-promoting factor found at elevated concentrations in the peritoneal fluid of women with endometriosis and active endometriosis lesions, acts directly on ectopic endometrial cells to stimulate the synthesis of COX-2, the inducible form of COX, and the release of PGE(2). MIF treatment strongly activated p38 and ERK MAPK, and specific inhibitors of both pathways completely blocked basal and MIF-induced PGE(2) synthesis. Whereas p38 inhibitors negatively affected the stimulated synthesis of COX-2 and that of PGE(2), ERK inhibitors only decreased the production of PGE(2). These findings show for the first time a direct role for MIF in the up-regulation of COX-2 synthesis and PGE(2) secretion in ectopic endometrial cells. They further indicate that whereas p38 and ERK MAPK signaling pathways both play a significant role in the regulation of basal and MIF-induced synthesis of PGE(2) by ectopic endometrial cells, only p38 kinase is involved in the regulation of COX-2 expression in these cells. This suggests that MIF acts at more than one level to stimulate the synthesis of PGE(2) and triggers the coordinate activation of multiple enzymes in the biosynthesis pathway. Our data provide evidence for a novel mechanism by which MIF can induce a proinflammatory phenotype in ectopic endometrial cells, and favor the establishment of endometriosis and its related clinical symptoms.

An unrecognized extracellular function for an intracellular adapter protein released from the cytoplasm into the tumor microenvironment

Mammalian cell membranes provide an interface between the intracellular and extracellular compartments. It is currently thought that cytoplasmic signaling adapter proteins play no functional role within the extracellular tumor environment. Here, by selecting combinatorial random peptide libraries in tumor-bearing mice, we uncovered a direct, specific, and functional interaction between CRKL, an adapter protein [with Src homology 2 (SH2)- and SH3-containing domains], and the plexin-semaphorin-integrin domain of beta(1) integrin in the extracellular milieu. Through assays in vitro, in cellulo, and in vivo, we show that this unconventional and as yet unrecognized protein-protein interaction between a regulatory integrin domain (rather than a ligand-binding one) and an intracellular adapter (acting outside of the cells) triggers an alternative integrin-mediated cascade for cell growth and survival. Based on these data, here we propose that a secreted form of the SH3/SH2 adaptor protein CRKL may act as a growth-promoting factor driving tumorigenesis and may lead to the development of cancer therapeutics targeting secreted CRKL.

TGF-β Induces Growth Arrest in Burkitt Lymphoma Cells via Transcriptional Repression of E2F-1

Transforming growth factor-beta (TGF-beta) is a potent regulator of tissue homeostasis and can act as both a tumor suppressor and a tumor promoter. The ability to induce cell cycle arrest is a major component of the tumor suppressor function of TGF-beta. Lung, mammary, and skin epithelial cells exhibit a common minimal cytostatic program in response to TGF-beta signaling involving the repression of the growth-promoting factors c-MYC, Id1, Id2, and Id3. Loss of c-MYC expression is a pivotal event in this process, resulting in derepression of the cyclin-dependent kinase inhibitors CDKN1A (p21) and CDKN2B (p15) and ultimately leading to growth arrest. It is not clear, however, which responses are necessary for TGF-beta-mediated growth arrest in other cell types. Here, in human Burkitt lymphoma cells transformed by deregulated c-MYC expression, we demonstrate that efficient TGF-beta-induced cytostasis can occur despite both maintenance of c-MYC levels and a lack of p21 and p15 induction. TGF-beta treatment also results in induction, rather than repression, of Id1 and Id2 expression. In this context, growth arrest correlates with transcriptional repression of E2F-1, and overexpression of E2F-1 in Burkitt lymphoma cells largely overcomes the TGF-beta-mediated G(1) arrest phenotype. These data indicate that deregulation of c-MYC in lymphoma cells does not overcome the tumor suppressor function of TGF-beta and that repression of E2F-1 transcription is sufficient for the efficient induction of cytostasis.

Effect of Hepatocyte Growth-Promoting Factors on Myocardial Ischemia During Exercise in Patients With Severe Coronary Artery Disease

Hepatocyte growth-promoting factor (pHGF) has the greatest potential as a therapeutic agent for vascular growth factor. The aim of this study was to investigate the effect of pHGF on myocardial ischemia and exercise capacity in patients with severe coronary artery disease (CAD). Forty-nine patients were enrolled for a two week treatment period. Treadmill graded exercise tests with gas analysis were conducted before and after therapy. Serum levels of HGF were significantly elevated after therapy. The degrees of exercise-induced ST segment depression were decreased more significantly in the pHGF group. Similar differences were also found in the maximum heart rate and the maximum heart rate when the ST segment was depressed 1 mm while undergoing the treadmill graded exercise test. Both were increased more significantly in the pHGF group. Total exercise time, systolic blood pressure in the peak of exercise, the length of time that ST segment depression of 1 mm is needed, and total work all were increased in both groups after intervention. Furthermore, total exercise time and total work were increased more significantly in the pHGF group. The levels of HGF increased significantly after pHGF treatment. pHGF could favorably improve exercise-induced myocardial ischemia and enhance exercise capacity in patients with severe CAD.

Treatment of chronic hepatic cirrhosis with autologous bone marrow stem cells transplantation in rabbits

Objective To evalute the feasibility of treatment for rabbit model with hepatic cirrhosis by transplantation of autologous bone marrow-derived stem cells via the hepatic artery and evaluate the effect of hepatocyte growth-promoting factors (pHGF) in the treatment of stem cells transplantation to liver cirrhosis.To provide empirical study foundation for future clinical application.Methods Chronic hepatic cirrhosis models of rabbits were developed by subcutaneous injection with 50% CCl4 0.2 ml/kg.Twenty-five model rabbits were randomly divided into three experimental groups,stem cells transplant group (10),stem ceils transplant + pHGF group (10) and control group (5).Autologous bone marrow was harvested from tibia of each rabbit,and stem cells were disassociated using density gradient centrifugation and transplanted into liver via the hepatic artery under fluoroscopic guidance.In the stem cells transplant + pHGF group,the hepatocyte growth-promoting factor was given via intravenous injection with 2 mg/kg every other day for 20 days.Liver function tests were monitored at 4,8,12 weeks intervals and histopathologic examinations were performed at 12 weeks following transplantation.The data were analyzed using analysis of variance Results Following transplantation of stem cells,the liver function of rabbits improved gradually.Twelve weeks after transplantation,the activity of ALT and AST decreased from (73.0±10,6)U/L and(152.4± 22.8) U/L to (48.0±1.0)U/L and(86.7±2.1)U/L respectively; and the level of ALB and PTA increased from (27.5 ±1.8)g/L and 28.3% to (33.2 +0.5)g/L and 44.1% respectively.The changes did not have statistically significant difference when compared to the control group(P >0.05).However,in the stern cellstransplant + pHGF group,the activity of ALT and AST decreased to (43.3±0.6)U/L and (78.7±4.0)U/L respectively and the level of ALB and PTA increased to (35.7 ±0.4)g/L and 50.5% respectively.The difference was statistically significant when compared to the control group(F=47.38,23.52,52.27,174.45,P cells within liver tissue in the stem cells trasplant group when compared to the control group,and the most hepatic CD34 cells were seen in the stem cells transplant + pHGF group.Conclusion Autologous bone marrow stem cells transplanted via the hepatic artery for the rabbit hepatic cirrhosis model is an effective method to treat hepatic cirrhosis.The hepatoeyte growth-promoting factor can help to enhance the results of treatment. Key words: Stem cell transplantation; Hepatic cirrhosis,experimental; Treatment outcome; Hepatic artery

Origins of Growth Factors: NGF and EGF

Growing up on the streets of Brooklyn, I remember being interested in how things worked: taking apart an old telephone and the gears of my new 4-speed bicycle was most enjoyable. As a biology major at Brooklyn College in 1945, I was fascinated by embryology. How does an egg turn into a chicken or a frog or a person? My only insight into the problem was the thought that it was necessary to understand the chemical reactions inside the egg and embryo and not simply observe biological structures. I, therefore, became a double major in chemistry and biology (also with the hope of earning a living as a laboratory technician). My first job after graduating was working nights as a bacteriologist in a milk processing plant. One of my professors at Brooklyn College wrote asking whether I was interested in going to graduate school. The college had been sending one student a year to the Biology Department at Oberlin to work as a laboratory assistant while studying for a master's degree. Because Oberlin offered to pay my tuition plus some $300 a semester for living expenses, I jumped at the chance. School and science were both interesting and fun. Upon graduating, I continued my education toward a Ph.D. working as an assistant in the Biochemistry Department of the University of Michigan under Howard B. Lewis. My research involved studying the Krebs urea cycle in the common earthworms that I collected from the university campus. I had been told at Oberlin that the yellow cells surrounding the gut in the worm corresponded to the mammalian liver. I decided to check whether the enzymatic reactions in worms were similar to those in mammals. They were (1). My first “real job” was in the Pediatrics Department at the University of Colorado studying creatine metabolism in premature infants under Harry Gordon. I think he hired me because he was impressed by my ability to stomach tube earthworms. After several years, I decided I needed to learn the then new technique of using radioisotopes in metabolic studies, and I obtained an American Cancer Society fellowship to work in the Radiology Department of Washington University under Martin Kamen where I combined this new knowledge with my interest in embryology. Although fertilized frog eggs and early embryos are impermeable to small molecules (amino acids, phosphates, etc.), sufficient amounts of C14O2 were metabolized by intact eggs and embryos to permit the identification of the radioactive compounds present at various stages of development (2). Upon completion of my fellowship, Dr. Kamen recommended me for a position in the Zoology Department of Washington University in the laboratory of Viktor Hamburger and Rita Levi-Montalcini. This move was critical in determining the direction of my research for the next 40-some odd years: the isolation, structure, and function of the first of the “growth factors,” nerve growth factor (NGF) and epidermal growth factor (EGF).

Immune Response of Human Propagated γδ-T-Cells to Neuroblastoma Recommend the Vδ1+ Subset for γδ-T-cell–based Immunotherapy

Human peripheral gammadelta-T-cells are able to induce cytolysis of neuroblastoma (Nb) tumor cells. Besides innate effector functions against infected cells and tumors, gammadelta-T-cells are involved in T-helper 1/T-helper 2 (TH1/TH2) differentiation of alphabeta-T-cells. However, as different gammadelta-T-cell subsets vary considerably in their functional properties, the aim of the present study was to define repertoires of cytokines, chemokines, and angiogenic factors of in vitro expanded Vdelta1+ and Vdelta2+ T cells in response to Nb. After short-term culture, both subsets released TH1 [interleukin (IL)-2, interferon (IFN)-gamma, IL-12, tumor necrosis factor (TNF)-alpha, TNF-beta)] and TH2 cytokines (IL-4, -5, -6, -10, -13, Vdelta1 also transforming growth factor (TGF)-beta, chemokines (I-309, monocyte chemotactic protein (MCP)-1-3, regulated upon activation, normal T-cell expressed and secreted), ILs (IL-1, -8, -15), cytokines (leptin) as well as angiogenic growth factors [angiogenin (ANG), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), Insulin-like growth factor (IGF)-I]. These molecules were expressed at higher levels in Vdelta2+ than Vdelta1+ T cells. Nb challenge changed protein expression. TH2 cytokine and IFN-gamma release was blocked in both gammadelta-T-cell subsets. In Vdelta2 gammadelta-T-cells, TH1 cytokines were down-regulated and tumor growth-promoting factors (ANG, VEGF, EGF, and IGF-I) were strongly up-regulated. In contrast, Vdelta1+ gammadelta-T-cells stopped the release of tumor-supportive factors and tolerogenic TGF-beta, and strongly up-regulated TNF-alpha, TNF-beta, MCP-1 and -2 and maintained their IL-2 production. In summary, our data show that after being challenged with Nb cells, propagated Vdelta1+ rather than Vdelta2+ T cells support antitumor responses by secretion of proinflammatory cytokines. Furthermore, in contrast to other cell types, Vdelta1+ T cells do not sustain a growth-promoting or tolerogenic microenvironment. These data make Vdelta1+ T cells an ideal candidate for upcoming immunotherapy trials in Nb.

Inflammatory Pathways Are Activated during Cardiomyocyte Hypertrophy and Attenuated by Peroxisome Proliferator-activated Receptors PPARα and PPARδ

Accumulating evidence indicates an important role for inflammation in cardiac hypertrophy and failure. Peroxisome proliferator-activated receptors (PPARs) have been reported to attenuate inflammatory signaling pathways and, as such, may interfere with cardiac remodeling. Accordingly, the objectives of the present study were to explore the relationship between cardiomyocyte hypertrophy and inflammation and to investigate whether PPARalpha and PPARdelta are able to inhibit NF-kappaB activation and, consequently, the hypertrophic growth response of neonatal rat cardiomyocytes (NCM). mRNA levels of markers of both hypertrophy and inflammation were increased following treatment with the pro-hypertrophic factor phenylephrine (PE) or the chemokine TNF-alpha. Induction of inflammatory genes was found to be fast (within 2 h after stimulation) and transient, while induction of hypertrophic marker genes was more gradual (peaking at 24-48 h). Inflammatory and hypertrophic pathways appeared to converge on NF-kappaB as both PE and TNF-alpha increased NF-kappaB binding activity as measured by electrophoretic mobility shift assay. Following transient transfection, the p65-induced transcriptional activation of a NF-kappaB reporter construct was significantly blunted after co-transfection of PPARalpha or PPARdelta in the presence of their respective ligands. Finally, adenoviral overexpression of PPARalpha and PPARdelta markedly attenuated cell enlargement and the expression of hypertrophic marker genes in PE-stimulated NCM. The collective findings reveal a close relationship between hypertrophic and inflammatory signaling pathways in the cardiomyocyte. It was shown that both PPARalpha and PPARdelta are able to mitigate cardiomyocyte hypertrophy in vitro by inhibiting NF-kappaB activation.

Stromal myofibroblasts are drivers of invasive cancer growth

Tissue integrity is maintained by the stroma in physiology. In cancer, however, tissue invasion is driven by the stroma. Myofibroblasts and cancer-associated fibroblasts are important components of the tumor stroma. The origin of myofibroblasts remains controversial, although fibroblasts and bone marrow-derived precursors are considered to be the main progenitor cells. Myofibroblast reactions also occur in fibrosis. Therefore, we wonder whether nontumorous myofibroblasts have different characteristics and different origins as compared to tumor-associated myofibroblasts. The mutual interaction between cancer cells and myofibroblasts is dependent on multiple invasive growth-promoting factors, through direct cell-cell contacts and paracrine signals. Since fibrosis is a major side effect of radiotherapy, we address the question how the main methods of cancer management, including chemotherapy, hormonotherapy and surgery affect myofibroblasts and by inference the surrogate endpoints invasion and metastasis.

Macrophage migration inhibitory factor (MIF)-induced signaling pathways in human endometrial stromal cells of women with and without endometriosis

OBJECTIVE: To investigate signal transduction pathways in endometrial stromal cells of women with and without endometriosis in response to MIF, a potent proinflammatory and growth-promoting factor.DESIGN: Primary cultures of endometrial stromal cells isolated from endometrial biopsies of women with and without endometriosis in proliferative and secretory cycle phase exposed to MIF and assessment of extracellular signal-related kinase (Erk)1/2 mitogen-activated protein kinase (MAPK), p38 MAPK and Akt phosphorylation.MATERIALS AND METHODS: Cells were cultured in DMEM-F12 medium with 10% FBS, 1% antibiotics, 5 μg/mL insulin and 5 μg/mL transferrin. Cells grown to confluence were preincubated for 1 h with Erk1/2 MAPK, p38 MAPK and Akt inhibitors (PD98059, SB203580 and LY294002 respectively) and stimulated with different concentrations of MIF (0-50 ng/mL) for different periods of time (0-1 h). Then, the extraction of proteins was performed and phosphorylation of Erk1/2 MAPK, p38 MAPK and Akt was analyzed by Western blot. An unpaired t test and one-way ANOVA followed by the Dunnett's test were performed for this study.RESULTS: Our study showed that MIF stimulated p38 MAPK and Akt phosphorylation in endometrial cells of women with and without endometriosis in the proliferative and the secretory phases of the menstrual cycle. In normal women, the basal level of Erk1/2 MAPK phosphorylation was lower in cells from the secretory phase of the menstrual cycle compared to the proliferative phase, but remained steadily elevated throughout the menstrual cycle in women with endometriosis. Cell exposure to MIF did not result in any detectable increase in Erk1/2 MAPK phosphorylation, whereas in normal women MIF activated Erk1/2 MAPK phosphorylation in the secretory phase. PD98059, SB203580 and LY294002 suppressed the ability of MIF to increase Erk1/2 MAPK, p38 MAPK and Akt phosphorylation, respectively.CONCLUSIONS: p38 and Akt signaling pathways are involved in MIF-mediated activation of endometrial stromal cells of women with and without endometriosis. Activation of Erk1/2 in endometrial cells throughout the menstrual cycle of women with endometriosis may, considering the role of these kinases in cell growth, angiogenesis and tissue remodeling, play an important role in the capability of endometrial cells to implant and develop in ectopic locations. OBJECTIVE: To investigate signal transduction pathways in endometrial stromal cells of women with and without endometriosis in response to MIF, a potent proinflammatory and growth-promoting factor. DESIGN: Primary cultures of endometrial stromal cells isolated from endometrial biopsies of women with and without endometriosis in proliferative and secretory cycle phase exposed to MIF and assessment of extracellular signal-related kinase (Erk)1/2 mitogen-activated protein kinase (MAPK), p38 MAPK and Akt phosphorylation. MATERIALS AND METHODS: Cells were cultured in DMEM-F12 medium with 10% FBS, 1% antibiotics, 5 μg/mL insulin and 5 μg/mL transferrin. Cells grown to confluence were preincubated for 1 h with Erk1/2 MAPK, p38 MAPK and Akt inhibitors (PD98059, SB203580 and LY294002 respectively) and stimulated with different concentrations of MIF (0-50 ng/mL) for different periods of time (0-1 h). Then, the extraction of proteins was performed and phosphorylation of Erk1/2 MAPK, p38 MAPK and Akt was analyzed by Western blot. An unpaired t test and one-way ANOVA followed by the Dunnett's test were performed for this study. RESULTS: Our study showed that MIF stimulated p38 MAPK and Akt phosphorylation in endometrial cells of women with and without endometriosis in the proliferative and the secretory phases of the menstrual cycle. In normal women, the basal level of Erk1/2 MAPK phosphorylation was lower in cells from the secretory phase of the menstrual cycle compared to the proliferative phase, but remained steadily elevated throughout the menstrual cycle in women with endometriosis. Cell exposure to MIF did not result in any detectable increase in Erk1/2 MAPK phosphorylation, whereas in normal women MIF activated Erk1/2 MAPK phosphorylation in the secretory phase. PD98059, SB203580 and LY294002 suppressed the ability of MIF to increase Erk1/2 MAPK, p38 MAPK and Akt phosphorylation, respectively. CONCLUSIONS: p38 and Akt signaling pathways are involved in MIF-mediated activation of endometrial stromal cells of women with and without endometriosis. Activation of Erk1/2 in endometrial cells throughout the menstrual cycle of women with endometriosis may, considering the role of these kinases in cell growth, angiogenesis and tissue remodeling, play an important role in the capability of endometrial cells to implant and develop in ectopic locations.