scholarly journals Caffeine Inhibits EGF-Stimulated Trophoblast Cell Motility through the Inhibition of mTORC2 and Akt

Endocrinology ◽  
2012 ◽  
Vol 153 (9) ◽  
pp. 4502-4510 ◽  
Author(s):  
Isobelle Grant ◽  
Judith E. Cartwright ◽  
Brooke Lumicisi ◽  
Alison E. Wallace ◽  
Guy S. Whitley
Keyword(s):  
Cell Motility ◽  
Signaling Pathways ◽  
Intracellular Signaling ◽  
Pregnancy Loss ◽  
Mammalian Target Of Rapamycin ◽  
First Trimester ◽  
Trophoblast Invasion ◽  
Intracellular Signaling Pathways

Impaired trophoblast invasion is associated with pregnancy disorders such as early pregnancy loss and preeclampsia. There is evidence to suggest that the consumption of caffeine during pregnancy may increase the risk of pregnancy loss; however, little is known about the direct effect of caffeine on normal trophoblast biology. Our objectives were to examine the effect of caffeine on trophoblast migration and motility after stimulation with epidermal growth factor (EGF) and to investigate the intracellular signaling pathways involved in this process. Primary first-trimester extravillous trophoblasts (EVT) and the EVT-derived cell line SGHPL-4 were used to study the effect of caffeine on EGF-stimulated cellular motility using time-lapse microscopy. SGHPL-4 cells were further used to study the effect of caffeine and cAMP on EGF-stimulated invasion of fibrin gels. The influence of caffeine and cAMP on EGF-stimulated intracellular signaling pathways leading to the activation of Akt were investigated by Western blot analysis. Caffeine inhibits both EGF-stimulated primary EVT and SGHPL-4 cell motility. EGF stimulation activates phosphatidylinositol 3-kinase, and Akt and caffeine inhibit this activation. Although cAMP inhibits both motility and invasion, it does not inhibit the activation of Akt, indicating that the effects of caffeine seen in this study are independent of cAMP. Further investigation indicated a role for mammalian target of rapamycin complex 2 (mTORC2) as a target for the inhibitory effect of caffeine. In conclusion, we demonstrate that caffeine inhibits EGF-stimulated trophoblast invasion and motility in vitro and so could adversely influence trophoblast biology in vivo.

scholarly journals Preeclampsia-related increase of interleukin-11 expression in human decidual cells

Reproduction ◽  
2010 ◽  
Vol 140 (4) ◽  
pp. 605-612 ◽  
Author(s):  
Murat Basar ◽  
Chih-Feng Yen ◽  
Lynn F Buchwalder ◽  
William Murk ◽  
S Joseph Huang ◽  
...  
Keyword(s):  
Signaling Pathways ◽  
P38 Mapk ◽  
First Trimester ◽  
P38 Map Kinase ◽  
Trophoblast Invasion ◽  
Decidual Cells ◽  
Pkc Signaling ◽  
Regulatory Effects

Preeclampsia is associated with increased systemic inflammation and superficial trophoblast invasion, which leads to insufficient uteroplacental blood flow. Interleukin (IL)-11 mediates pro- and anti-inflammatory processes and facilitates decidualization. To identify IL11 expression in vivo at the maternal–placental interface in preeclampsia and control specimens and to evaluate the regulatory effects of tumor necrosis factor-α (TNF) and IL1B, cytokines elevated in preeclampsia, on IL11 levels in first trimester decidual cells in vitro, placental sections were immunostained for IL11. Leukocyte-free first trimester decidual cells were incubated with estradiol (E2)±10−7 mol/l medroxyprogesterone acetate±TNF or IL1B± inhibitors of the p38 MAP kinase (p38 MAPK), nuclear factor-κ B (NFKB), or protein kinase C (PKC) signaling pathways. An ELISA assessed secreted IL11 levels, and quantitative RT-PCR measured IL11 mRNA. IL11 immunoreactivity in placental sections was significantly higher in the cytoplasm of preeclamptic decidual cells versus gestational age-matched controls. Compared to decidual cells, IL11 immunostaining in neighboring trophoblast is lower, perivascular, and not different between control and preeclamptic specimens. TNF and IL1B enhanced levels of IL11 mRNA and secreted IL11 in cultured decidual cells. Specific inhibitors of the p38 MAPK and NFKB, but not PKC signaling pathways, reduced the stimulatory effect of IL1B. Expression of decidual IL11 is increased in preeclampsia and suggests a role for IL11 in the pathogenesis of preeclampsia.

scholarly journals Study of intracellular signaling pathways in Chronic Myeloproliferative Neoplasms

2018 ◽  
Author(s):  
Serena Martinelli
Keyword(s):  
Signaling Pathways ◽  
Intracellular Signaling ◽  
Myeloproliferative Neoplasms ◽  
Mtor Pathway ◽  
Janus Kinase ◽  
Pi3k Inhibitor ◽  
Chronic Myeloproliferative Neoplasms ◽  
Intracellular Signaling Pathways

A gain-of-function mutation in Janus kinase 2 (JAK2V617F) is at the basis of the majority of chronic myeloproliferative neoplasms (MPN). Enhanced activation of other downstream pathways including the PI3K/mTOR pathway has been documented as well. In this study we evaluated the effects of JAK1/2 inhibitors, alone and in combination with mTOR, with a dual mTOR/PI3K inhibitor and with a pan PI3K inhibitor in in-vitro and in-vivo MPN models. Our findings of strong synergy between the JAK2 inhibitors and mTOR/PI3K inhibitor suggested that we might be able to administer these drugs at lower concentrations than when the drugs are used individually. This provides a framework for combination trials using compounds in patients with myeloproliferative neoplasms

Cell Motility in Microfabricated Models of the Tissue Microenvironment

2001 ◽  
Author(s):  
Kevin Kit Parker ◽  
Donald E. Ingber
Keyword(s):  
Cell Motility ◽  
Signaling Pathways ◽  
Intracellular Signaling ◽  
Isometric Tension ◽  
Migration Behavior ◽  
Tissue Microenvironment ◽  
Intracellular Signaling Pathways ◽  
Capillary Endothelial Cells ◽  
Geometric Confinement ◽  
Stable Rotation

Abstract We conducted studies using micropatterned substrates to elucidate how cell shape and geometric confinement regulates the inter- and intracellular signaling pathways required for cell motility. When cells were cultured on individual cell-sized square adhesive islands coated with ECM, they extend to the edge of the island and assume a square shape. When these cells were stimulated with growth factors, they preferentially extended lamellipodia from the corners versus the sides. This process was mediated by myosin-generated isometric tension that induced tight spatial localization of Rac in the corners. When two or three capillary endothelial cells are constrained to a fibronectin (FN) island, coordinated cell migration results in stable rotation of the entire system about its center. Thus, the emergent pattern is due to the coordinated migration behavior of the cells. These observations suggest that ECM-induced mechanotransduction potentiates compartmentalized signaling pathways and the multicellular organization required of tissue morphogenesis.

Curcumin inhibits cystogenesis by simultaneous interference of multiple signaling pathways: in vivo evidence from aPkd1-deletion model

2011 ◽  
Vol 300 (5) ◽  
pp. F1193-F1202 ◽  
Author(s):  
Wouter N. Leonhard ◽  
Annemieke van der Wal ◽  
Zlata Novalic ◽  
Steven J. Kunnen ◽  
Ron T. Gansevoort ◽  
...  
Keyword(s):  
Renal Failure ◽  
Signaling Pathways ◽  
Early Stage ◽  
Mammalian Target Of Rapamycin ◽  
Therapeutic Interventions ◽  
Proliferation Index ◽  
Cystic Kidneys ◽  
Renal Histology

Autosomal dominant polycystic kidney disease (ADPKD) caused by mutations in either the PKD1 or PKD2 gene is a major cause of end-stage renal failure. A number of compounds targeting specific signaling pathways were able to inhibit cystogenesis in rodent models and are currently being tested in clinical trials. However, given the complex signaling in ADPKD, an ideal therapy would likely have to comprise several pathways at once. Therefore, multitarget compounds may provide promising therapeutic interventions for the treatment of ADPKD. To test this hypothesis, we treated Pkd1-deletion mice with diferuloylmethane (curcumin), a compound without appreciable side effects and known to modulate several pathways that are also altered in ADPKD, e.g., mammalian target of rapamycin (mTOR) and Wnt signaling. After conditional inactivation of Pkd1, mTOR signaling was indeed elevated in cystic kidneys. Interestingly, also activation of signal transducers and activator of transcription 3 (STAT3) strongly correlated with cyst progression. Both pathways were effectively inhibited in vitro by curcumin. Importantly, Pkd1-deletion mice that were treated with curcumin and killed at an early stage of PKD displayed improved renal histology and reduced STAT3 activation, proliferation index, cystic index, and kidney weight/body weight ratios. In addition, renal failure was significantly postponed in mice with severe PKD. These data suggest that multitarget compounds hold promising potential for safe and effective treatment of ADPKD.

CD45 Phosphatase Is Involved in Motility and Development of Hematopoietic Stem and Maturing Cells by the Regulation of Cell Adhesion and Cytokine Signaling.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 119-119
Author(s):  
Shoham Shivtiel ◽  
Isabelle Petit ◽  
Abraham Avigdor ◽  
Polina Goichberg ◽  
Sarit Samira ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Cell Motility ◽  
Signaling Pathways ◽  
Stem Cell Differentiation ◽  
Hematopoietic Cell ◽  
Cytokine Signaling ◽  
Hematopoietic Stem ◽  
Migration And Development

Abstract The phosphatase CD45 is a key regulator of antigen receptor signaling in lymphocytes. Still, CD45 is highly expressed in all hematopoietic lineages at most stages of development, suggesting that this phosphatase also regulates other cells and processes. During development, as well as in clinical transplantation, hematopoietic stem cells (HSCs) migrate through the circulation to the bone marrow (BM) and repopulate it. Migration and development of HSCs are multi-step processes, which are tightly regulated by interplays between cytokines, chemokines, adhesion molecules and proteolytic enzymes; however, not all the related key players have been fully identified. In this study we explored the involvement of CD45 in hematopoietic cell motility and development, its role in cytokine signaling and adhesion interactions. The roles of CD45 were tested by either blocking the function of CD45 expressed on human and murine HSCs by neutralizing antibodies, or by utilizing CD45 knockout (KO) mice. Our results show that blocking CD45 completely prevented homing of human CD34+ enriched progenitors to the murine BM, consequently abrogating repopulation in transplanted NOD/SCID mice. In addition, CD45 neutralization impaired the capacity of human progenitors to migrate in-vitro towards a gradient of SDF-1, suggesting a cross-talk between SDF-1 and CD45 signaling. Furthermore, blocking CD45 on human G2 cells (pre-B ALL line) activated signaling pathways, including an increase in phosphorylation of MAP kinase and the tyrosine kinase Pyk2, which are involved in cell adhesion and migration. This activation enhanced cell adhesion to stromal and endothelial cell lines in-vitro. Importantly, blockage of CD45 in human progenitors resulted in cell aggregation, which inhibited cell proliferation and impaired the capacity to form colonies in-vitro. In an additional set of experiments we tested the role of CD45 in cell mobilization. In-vivo studies in normal mice demonstrated that neutralization of CD45 function inhibited the release of mature leukocytes and progenitor cells from the BM to the circulation both under steady state conditions and in stress-induced recruitment by stimulation with G-CSF or LPS. More importantly, BM derived mononuclear cells from CD45KO mice displayed a significant reduction in in-vivo homing and in-vitro migration compared to their wild type counterparts. Furthermore, G-CSF induced mobilization was impaired in CD45KO mice, and accompanied by a reduction in MMP-9 secretion from blood-derived leukocytes. Unexpectedly, the ability of CD45KO progenitors to form colonies in-vitro was impaired in the absence of in-vivo BM environment, documenting a crucial role for phosphatases such as CD45 in stem cell differentiation. Taken together, our findings demonstrate that functional CD45 is essential for human and murine hematopoietic cell migration and development (both homing and mobilization) by the regulation of adhesion and cytokine-induced signaling machineries. We suggest that in these cells CD45 may act as a negative regulator of major signaling pathways controlling adhesion properties and maintaining the balance between anchorage and release. We reveal a novel and dual role for CD45 in regulation of hematopoietic cell trafficking in general and progenitor cell motility and development in particular.

scholarly journals Repurposing Tranexamic Acid as an Anticancer Agent

2021 ◽  
Author(s):  
Mary E Law ◽  
Bradley J Davis ◽  
Amanda F Ghilardi ◽  
Elham Yaaghubi ◽  
Zaafir M Dulloo ◽  
...  
Keyword(s):  
Tranexamic Acid ◽  
Anticancer Agents ◽  
Intracellular Signaling ◽  
Anticancer Agent ◽  
Lead Compounds ◽  
Intracellular Signaling Pathways ◽  
Multiple Processes ◽  
Broad Array

Tranexamic Acid (TA) is a clinically used antifibrinolytic that acts as a lysine mimetic to block binding of Plasminogen with Plasminogen activators, preventing conversion of Plasminogen to its proteolytically activated form, Plasmin. Previous studies suggested that TA may exhibit anticancer activity by blockade of extracellular Plasmin formation. Plasmin-mediated cleavage of the CDCP1 protein may increase its oncogenic functions through several downstream pathways. Results presented herein demonstrate that TA blocks Plasmin-mediated excision of the extracellular domain of the oncoprotein CDCP1. In vitro studies indicate that TA reduces the viability of a broad array of human and murine cancer cell lines, and breast tumor growth studies demonstrate that TA reduces cancer growth in vivo. Based on the ability of TA to mimic lysine and arginine, we hypothesized that TA may perturb multiple processes that involve Lys/Arg-rich protein sequences, and that TA may alter intracellular signaling pathways in addition to blocking extracellular Plasmin production. Indeed, TA-mediated suppression of tumor cell viability is associated with multiple biochemical actions, including inhibition of protein synthesis, reduced activating phosphorylation of STAT3 and S6K1, decreased expression of the MYC oncoprotein, and suppression of Lys acetylation. These findings suggest that TA or TA analogs may serve as lead compounds and inspire the production of new classes of anticancer agents that function by mimicking Lys and Arg.

Analysis of Differentially Activated Signaling Pathways in Myeloproliferative Disease Using Kinomics Chip Profiling

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 3723-3723
Author(s):  
Daniel B Lipka ◽  
Marie-Christine Blum ◽  
Florian H Heidel ◽  
Thomas Kindler ◽  
Thomas Fischer
Keyword(s):  
Signaling Pathways ◽  
Cell Line ◽  
Intracellular Signaling ◽  
Jak2v617f Mutation ◽  
Myeloproliferative Disease ◽  
Differential Activation ◽  
Cell Lysate ◽  
Intracellular Signaling Pathways ◽  
Chip Analysis

Abstract In a multitude of cases, oncogenic mutations are gain of function mutations that confer a constitutively activated gene product. Currently, evidence from a large body of experimental studies suggests that oncogenic transformation induced by activating kinase mutations is not sufficiently explained by constitutive kinase activation alone but is a result of aberrantly activated signaling pathways in affected cells. The JAK2V617F-mutation is a highly prevalent molecular marker in Ph-negative myeloproliferative disease (MPD). In vitro, Ba/F3-cells expressing both erythropoietin receptor (EpoR) and the JAK2V617F-mutation show constitutive activation of the JAK-STAT pathway and cytokine independent growth. Multiple in-vitro and in-vivo studies demonstrate that the JAK2V617F-mutation mediates many of the phenotypic characteristics of the MPDs. Nevertheless, until now it is largely unclear, which signaling pathways in particular are involved in the process of malignant transformation in JAK2V617F-positive cells. Therefore, we applied a kinomics chip approach to screen for activated intracellular signaling pathways. We used a commercially available peptide chip containing 960 synthetic kinase substrate peptides spotted in triplicates and covering peptide substrates for approximately 50% of the human kinome. Peptides have been selected for their biologic relevance in physiologic processes such as stress response, growth and cell differentiation. With this approach, a broad spectrum of intracellular signaling pathways and kinases can be investigated simultaneously in a single experiment. As a proof of principle, we performed kinomics chip analysis of Ba/F3-cells stably transfected with EpoR and either the JAK2V617F-mutant (Ba/F3-EpoR-VF) or wildtype JAK2 (Ba/F3-EpoR-WT). In brief, cells were seeded and treated with erythropoietin. One chip per cell lysate was incubated with an activation buffer containing the cell lysate and radioactively labelled ATP for a defined time period and washed several times afterwards. The chip was then analyzed by means of autoradiography using a phospho-storage-screen and a phospho-imager. Chip analysis was performed using standard microarray software and Microsoft Excel software. Chip experiments were performed simultaneously for Ba/F3-EpoR-VF and Ba/F3-EpoR-WT and in duplicate. Analysis revealed differential activation of known pathways such as Ras/Raf/MEK/ERK, JAK/STAT, and PI3-Kinase with pronounced activation seen in Ba/F3-EpoR-VF cells as compared to Ba/F3-EpoR-WT cells. This was not a surprising result but strongly underlines the feasibility and validity of this approach and therefore served as an internal control. Differential regulation of a number of other signaling nodes that have not yet been described in the context of mutant JAK2 signaltransduction have been detected. To select for relevant hits among these potential targets, we first excluded all substrates from further analysis that are known to be involved in lymphocyte-specific pathways. For the remaining hits we performed a literature search to learn more about their known functions and their potential impact in JAK2V617F-positive MPD. Validation of selected signaling molecules by means of Western blotting analysis and functional investigations such as siRNA knock-down experiments are currently under way. In addition to the widely used lymphoid Ba/F3-model, we also established a novel cell culture model with simultaneous expression of EpoR and either mutant or wildtype JAK2 in a myeloid 32D-cell background. This model will be helpful to us to determine false-positive results due to cell-line specific changes. We conclude, that kinomic profiling using the above mentioned chip-technology is a valid method to comprehensively investigate differential activation of signaling pathways in cell lysates. In our cell line model, we were able to detect activation of well known signaling pathways in JAK2V617F-positive cells. Furthermore, we were able to identify candidate proteins that appear to be specifically involved in JAK2V617F-signaling.

scholarly journals Protosappanin B Exerts Anti-tumor Effects on Colon Cancer Cells via Inhibiting GOLPH3 Expression

2020 ◽  
Vol 19 ◽  
pp. 153473542097247
Author(s):  
Xue-Cong Zheng ◽  
Ze-Sheng Shi ◽  
Cheng-Zhi Qiu ◽  
Zhong-Shi Hong ◽  
Chun-Xiao Wang ◽  
...  
Keyword(s):  
Colon Cancer ◽  
Cancer Cells ◽  
Signaling Pathways ◽  
Intracellular Signaling ◽  
Dependent Manner ◽  
Colon Cancer Cells ◽  
Human Colon Cancer ◽  
Intracellular Signaling Pathways ◽  
Sw620 Cells

Protosappanin B (PSB) is a key active component of Lignum Sappan extract. Although the antiproliferative effects of Lignum Sappan extract have been demonstrated in various cancer cells, relatively little is known about the effects of PSB on tumor progression. The aim of this study was to explore the anti-tumor effects of PSB on human colon cancer cells by regulation of intracellular signaling pathways and Golgi phosphoprotein 3 (GOLPH3) expression in vitro and in vivo. Our results showed that PSB effectively inhibited the viability and migration of SW620 cells and induced apoptosis, but had poor effect on HCT116 cells. Furthermore, PSB significantly reduced the expression of p-AKT, p-p70S6K, β-catenin, and p-ERK1/2 proteins in SW620 cells, and this effect was reversed by the corresponding signaling pathway agonists. Interestingly, PSB could also suppress GOLPH3 expression of SW620 cells in a concentration-dependent manner, but SW620 cells transfected with lentiviral vectors overexpressing GOLPH3 can effectively resist the cytotoxic activity of PSB in vitro. The xenograft experiment of SW620 cells with LV-GOLPH3 confirmed that PSB distinctly inhibited the tumor growth via suppressing GOLPH3 expression. Collectively, these findings clarified a new anti-cancer mechanism of PSB through inhibition of GOLPH3 expression and intracellular signaling pathways in colon cancer cells. PSB may be a potential new drug for colon cancer.

scholarly journals Adhesive and degradative properties of human placental cytotrophoblast cells in vitro.

1989 ◽  
Vol 109 (2) ◽  
pp. 891-902 ◽  
Author(s):  
S J Fisher ◽  
T Y Cui ◽  
L Zhang ◽  
L Hartman ◽  
K Grahl ◽  
...  
Keyword(s):  
Basement Membrane ◽  
First Trimester ◽  
Late Gestation ◽  
Trophoblast Invasion ◽  
Third Trimester ◽  
Biochemical Basis ◽  
Teratocarcinoma Cell ◽  
Cytotrophoblast Cells

Human fetal development depends on the embryo rapidly gaining access to the maternal circulation. The trophoblast cells that form the fetal portion of the human placenta have solved this problem by transiently exhibiting certain tumor-like properties. Thus, during early pregnancy fetal cytotrophoblast cells invade the uterus and its arterial network. This process peaks during the twelfth week of pregnancy and declines rapidly thereafter, suggesting that the highly specialized, invasive behavior of the cytotrophoblast cells is closely regulated. Since little is known about the actual mechanisms involved, we developed an isolation procedure for cytotrophoblasts from placentas of different gestational ages to study their adhesive and invasive properties in vitro. Cytotrophoblasts isolated from first, second, and third trimester human placentas were plated on the basement membrane-like extracellular matrix produced by the PF HR9 teratocarcinoma cell line. Cells from all trimesters expressed the calcium-dependent cell adhesion molecule cell-CAM 120/80 (E-cadherin) which, in the placenta, is specific for cytotrophoblasts. However, only the first trimester cytotrophoblast cells degraded the matrices on which they were cultured, leaving large gaps in the basement membrane substrates and releasing low molecular mass 3H-labeled matrix components into the medium. No similar degradative activity was observed when second or third trimester cytotrophoblast cells, first trimester human placental fibroblasts, or the human choriocarcinoma cell lines BeWo and JAR were cultured on radiolabeled matrices. To begin to understand the biochemical basis of this degradative behavior, the substrate gel technique was used to analyze the cell-associated and secreted proteinase activities expressed by early, mid, and late gestation cytotrophoblasts. Several gelatin-degrading proteinases were uniquely expressed by early gestation, invasive cytotrophoblasts, and all these activities could be abolished by inhibitors of metalloproteinases. By early second trimester, the time when cytotrophoblast invasion rapidly diminishes in vivo, the proteinase pattern of the cytotrophoblasts was identical to that of term, noninvasive cells. These results are the first evidence suggesting that specialized, temporally regulated metalloproteinases are involved in trophoblast invasion of the uterus. Since the cytotrophoblasts from first trimester and later gestation placentas maintain for several days the temporally regulated degradative behavior displayed in vivo, the short-term cytotrophoblast outgrowth culture system described here should be useful in studying some of the early events in human placen

Sign in / Sign up

Export Citation Format

Share Document