DOCSLIB.ORG

Essential Role of Stat3 in PI3K-Induced Oncogenic Transformation

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Essential Role of Stat3 in PI3K-Induced Oncogenic Transformation Essential role of Stat3 in PI3K-induced oncogenic transformation Jonathan R. Harta, Lujian Liaob, John R. Yates IIIb, and Peter K. Vogta,1 aDepartment of Molecular and Experimental Medicine and bDepartment of Chemical Physiology, The Scripps Research Institute, La Jolla, CA 92037 Contributed by Peter K. Vogt, June 28, 2011 (sent for review March 2, 2011) Cells transformed by the p110α-H1047R mutant of PI3K show in- generate a positive autocrine feedback leading to activation of creased tyrosine phosphorylation of Stat3. This activation of Stat3 Stat3 (16). is important for the transformation process, because a dominant- The PI3K signaling pathway is part of the core regulatory negative mutant of Stat3 interferes with PI3K-induced oncogene- networks in the cell and affects virtually all cellular activities, sis. GDC-0941, a specific inhibitor of PI3K reduces the level of Stat3 including growth, replication, movement, differentiation, and phosphorylation. The effect of PI3K on Stat3 appears to be medi- metabolism. PI3K signaling is elevated in most human cancers. ated by a member of the Tec kinase family. The Tec kinase inhib- This aberrant activity can result from differential regulation of itor LFM-A13 blocks Stat3 phosphorylation in H1047R-transformed PI3K itself or members of the pathway; it can also be caused by cells. The Janus kinase inhibitor AG490 and the Src kinase inhibitor gain-of-function mutations in the pathway or loss-of-function Src-1, as well as rapamycin, have no effect on Stat3 phosphoryla- mutations in the PI3K antagonist PTEN. The PI3K pathway tion in H1047R-transformed cells. The H1047R-transformed cells connects to numerous other signaling nodes including Ras, p53, also release a factor that induces Stat3 phosphorylation in normal Hif1α, and Lkb1. However, crosstalk between PI3K and Stat cells with possible effects on the cellular microenvironment. In signaling has not been reported. Here we describe a unique link some human tumor cell lines, the enhanced phosphorylation of between Stat3 and PI3K. In PI3K-transformed cells, Stat3 is Stat3 is inhibited by both PI3K and by Tec kinase inhibitors, sug- activated. This activation is essential for the process of trans- fi gesting that the link between PI3K and Stat3 is signi cant in formation. Inhibition of PI3K prevents Stat3 phosphorylation, human cancer. and dominant-negative Stat3 interferes with PI3K-induced on- cogenic transformation. MEDICAL SCIENCES oncogenic signaling | tumor stroma | tyrosine kinase Results tat3 is a member of a transcription factor family that was Stat3 Is Activated in C3H 10T1/2 Mouse Fibroblasts Transformed by Sdiscovered during the analysis of IFN-induced transcription the PI3K Mutant p110α-H1047R. We have used stable isotope la- (1–6). Stats are transcriptional regulators controlled by a path- beling with amino acids in cell culture (SILAC) in conjunction way that can be activated by growth factor as well as cytokine with tandem mass spectrometry to analyze the changes to the receptors (7). Stat3 is activated in response to the epidermal global proteome induced by the expression of the oncogenic growth factor and to IL-6 (2). Activation by IL-6 is primarily H1047R mutant of p110α in C3H 10T1/2 cells (26). The up- mediated by receptor-associated kinases of the Janus kinase (Jak) regulated PI3K signaling in the H1047R-transformed cells is family, which phosphorylate cytoplasmic Stats at tyrosine, effect- documented in Fig. 1A. Several of the proteins up-regulated by ing dimerization, translocation into the nucleus, sequence-specific p110α-H1047R are known targets of Stats (Table 1) (26–32). The DNA binding, and transcriptional activation (8). corresponding genes contain IFN-stimulated response elements The role of Stats, however, goes far beyond the IFN response. or IFN-γ activation sites (GAS). These binding sites interact with Stat3 is an important and often essential factor in oncogenic the IFN-stimulated gene factor 3 complex which contains both cellular transformation and in cancer. It is a required target of IFN response factor and Stat proteins. We therefore investigated the Src oncoprotein (9). Expression of dominant-negative Stat3 possible activation of Stat proteins by phosphorylation (Fig. 1B). blocks Src-induced cellular transformation. Stat3 is frequently The activation of Stat proteins by phosphorylation was evaluated and persistently activated in a wide variety of cancers (10). Mu- by Western blotting. The p110α-H1047R-transformed 10T1/2 rine cells in which Stat3 has been genetically inactivated are re- cells show enhanced phosphorylation of Stat3 and Stat6 and a sistant to oncogenic transformation (11, 12). Constitutively active decrease of phosphorylation in Stat1 (Fig. 1B). Because of the mutants of Stat3 are sufficient to convert normal cells into cancer prominent role of Stat3 in cancer (9–13), we decided to inves- cells (13). tigate its significance in PI3K-induced oncogenic transformation. The canonical kinases of Stat are members of the Jak family. There is no tyrosine kinase in the canonical PI3K signaling However, several other tyrosine kinases can phosphorylate and pathway. However, activated TOR (target of rapamycin) can activate Stats, including both receptor tyrosine kinases, such as phosphorylate S727 of Stat3 (33, 34) and this phosphorylation Egfr (14), Fgfr (15), Met (16), and Erbb2 (17), and nonreceptor enhances the activity of Stat3. However, in the two 10T1/2 cell tyrosine kinases, such as the Src and Fak kinase families. Such lines, S727 is constitutively phosphorylated, regardless of the ex- noncanonical Stat kinases are activated either through mutation pression of p110α-H1047R (Fig. 1B). It is therefore unlikely that – or through the aberrant expression of cytokines (18 20). In the enhanced expression of Stat targets in PI3K-transformed chronic myelogenous leukemia, the BCR-ABL fusion kinase also cells is mediated by TOR. In the context of the following ex- mediates Stat3 phosphorylation, and the leukemic cells are de- pendent upon this activity for sustained proliferation (21). Activation of Stat transcription factors induces a variety of Author contributions: J.R.H., J.R.Y., and P.K.V. designed research; J.R.H., L.L., J.R.Y., and proliferative and prosurvival proteins as they suppress immune P.K.V. performed research; J.R.H., L.L., and J.R.Y. contributed new reagents/analytic tools; responses (22). Stat proteins enhance the expression of the J.R.H., L.L., J.R.Y., and P.K.V. analyzed data; and J.R.H. and P.K.V. wrote the paper. antiapoptotic Bcl2 and Bcl-XL (13) and repress the expression of The authors declare no conflict of interest. proapoptotic proteins, such as p53 (23). Many growth factors are 1To whom correspondence should be addressed. E-mail: [email protected]. under Stat3 transcriptional control, including VEGF (24) and This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. HGF (25). Constitutive expression of these growth factors can 1073/pnas.1110486108/-/DCSupplemental. www.pnas.org/cgi/doi/10.1073/pnas.1110486108 PNAS Early Edition | 1of6 Downloaded by guest on October 2, 2021 phosphorylation of Y705 on Stat3 indicating that PI3K activity is essential for this phosphorylation event. Cells Expressing p110α-H1047R Release a Stat3-Activating Factor. The reduction of Stat3 phosphorylation by a PI3K inhibitor does not rule out an IFN response to the retroviral vector. To explore a potential role of IFN further, we transfected 10T1/2 cells with the TLR3/RIG-I agonist poly-I:C to trigger an authentic IFN response. Cell-free conditioned medium from these cells and from nontransfected H1047R-expressing cells was then used to stimulate 10T1/2 cells. The cells were lysed 2 h after exposure to the conditioned medium and analyzed by Western blot. IFN produced in response to poly-I:C transfection triggered a large increase in the expression of Isg15, but did not affect phos- phorylation of Stat3 (Fig. 1D). In contrast, medium conditioned by H1047R-transformed cells strongly stimulated the phosphor- ylation of Stat3, but had no effect on the production of Isg15. These data suggest that H1047R-transformed cells do not re- lease detectable quantities of IFN, but release a factor that can stimulate the phosphorylation of Stat3. The nature of this factor Fig. 1. (A) PI3K signaling in cells transformed by the H1047R mutant of is not known but is currently under investigation. p110α. The Western blot shows expression of p110α and phosphorylation of Akt and of S6 in the presence and in the absence of serum. H1047R-trans- Dominant-Negative Mutant of Stat3 Interferes with PI3K-Induced formed cells show constitutive activation of PI3K signaling in the absence of Oncogenic Transformation. To determine the functional signifi- serum. (B) Phosphorylation of Stat proteins. 10T1/2 and 10T1/2-H1047R cells cance of Stat3 activation in H1047R-induced transformation, we were lysed and probed for the indicated tyrosine phosphorylations of Stat examined a dominant negative, DNA binding-defective form of proteins. Transformation by p110α-H1047R induces an increase in Stat3 and Stat3 for its ability to affect the oncogenic potency of p110α- Stat6 phosphorylation and a decrease in Stat1 phosphorylation. S727 of fi Stat3, a known TOR phosphorylation site, is unaffected. (C) The PI3K in- H1047R. Chicken embryonic broblasts were transfected with hibitor GDC-0941 blocks tyrosine phosphorylation of Stat3. The 10T1/2- RCAS(B)-Stat3DB (9) or RCAS(B) as a vector control. After H1047R cells were treated with 20 μM GDC-0941 for 0, 1, 6, 24, or 48 h as 6 d to allow full expression of the dominant-negative Stat3, the indicated and then analyzed by Western blot. (D) IFN stimulation. 10T1/2 cultures were superinfected with serial dilutions of RCAS(A) ex- cells were exposed to conditioned medium from either 10T1/2-H1047R or pressing the oncogenic H1047R mutant of p110α or with ASV17 from normal 10T1/2 cells transfected with poly-I:C.
Load more

Recommended publications
  • Stem Cell Factor Is Selectively Secreted by Arterial Endothelial Cells in Bone Marrow
    ARTICLE DOI: 10.1038/s41467-018-04726-3 OPEN Stem cell factor is selectively secreted by arterial endothelial cells in bone marrow Chunliang Xu1,2, Xin Gao1,2, Qiaozhi Wei1,2, Fumio Nakahara 1,2, Samuel E. Zimmerman3,4, Jessica Mar3,4 & Paul S. Frenette 1,2,5 Endothelial cells (ECs) contribute to haematopoietic stem cell (HSC) maintenance in bone marrow, but the differential contributions of EC subtypes remain unknown, owing to the lack 1234567890():,; of methods to separate with high purity arterial endothelial cells (AECs) from sinusoidal endothelial cells (SECs). Here we show that the combination of podoplanin (PDPN) and Sca-1 expression distinguishes AECs (CD45− Ter119− Sca-1bright PDPN−) from SECs (CD45− Ter119− Sca-1dim PDPN+). PDPN can be substituted for antibodies against the adhesion molecules ICAM1 or E-selectin. Unexpectedly, prospective isolation reveals that AECs secrete nearly all detectable EC-derived stem cell factors (SCF). Genetic deletion of Scf in AECs, but not SECs, significantly reduced functional HSCs. Lineage-tracing analyses suggest that AECs and SECs self-regenerate independently after severe genotoxic insults, indicating the per- sistence of, and recovery from, radio-resistant pre-specified EC precursors. AEC-derived SCF also promotes HSC recovery after myeloablation. These results thus uncover heterogeneity in the contribution of ECs in stem cell niches. 1 The Ruth L. and David S. Gottesman Institute for Stem Cell and Regenerative Medicine Research, Albert Einstein College of Medicine, New York, NY 10461, USA. 2 Department of Cell Biology, Albert Einstein College of Medicine, New York, NY 10461, USA. 3 Department of Systems and Computational Biology, Albert Einstein College of Medicine, New York, NY 10461, USA.
    [Show full text]
  • Survival Pathways That Regulate Macrophage Tec and Btk in M-CSF
    Essential Roles for the Tec Family Kinases Tec and Btk in M-CSF Receptor Signaling Pathways That Regulate Macrophage Survival This information is current as of September 26, 2021. Martin Melcher, Bernd Unger, Uwe Schmidt, Iiro A. Rajantie, Kari Alitalo and Wilfried Ellmeier J Immunol 2008; 180:8048-8056; ; doi: 10.4049/jimmunol.180.12.8048 http://www.jimmunol.org/content/180/12/8048 Downloaded from References This article cites 36 articles, 18 of which you can access for free at: http://www.jimmunol.org/content/180/12/8048.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on September 26, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2008 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Essential Roles for the Tec Family Kinases Tec and Btk in M-CSF Receptor Signaling Pathways That Regulate Macrophage Survival1 Martin Melcher,2* Bernd Unger,2† Uwe Schmidt,3* Iiro A.
    [Show full text]
  • Redox-Mediated Regulation of the Tyrosine Kinase Zap70
    Redox-mediated regulation of the tyrosine kinase Zap70 DISSERTATION zur Erlangung des akademischen Grades doctor rerum naturalium (Dr. rer. nat.) genehmigt durch die Fakultät für Naturwissenschaften der Otto-von-Guericke-Universität von M.Sc. Christoph Thurm geb. am 27.06.1988 in Borna Gutachter: apl. Prof. Dr. Luca Simeoni PD Dr. rer. nat. Marcus Lettau eingereicht am: 02.02.2018 verteidigt am: 06.06.2018 Eigenständigkeitserklärung I. Eigenständigkeitserklärung Christoph Thurm Halberstädter Straße 29 39112 Magdeburg Hiermit erkläre ich, dass ich die von mir eingereichte Dissertation zu dem Thema Redox-mediated regulation of the tyrosine kinase Zap70 selbständig verfasst, nicht schon als Dissertation verwendet habe und die benutzten Hilfsmittel und Quellen vollständig angegeben wurden. Weiterhin erkläre ich, dass ich weder diese noch eine andere Arbeit zur Erlangung des akademischen Grades doctor rerum naturalium (Dr. rer. nat.) an anderen Einrichtungen eingereicht habe. Magdeburg, den 02.02.2018 ____________________________________ M.Sc. Christoph Thurm II ACKNOWLEDGEMENTS II. Acknowledgements Firstly, I would like to express my sincere gratitude to my supervisor Prof. Dr. Luca Simeoni. His extraordinary support during my PhD thesis together with his motivation and knowledge enabled me to pursue my dream. I could not have imagined having a better mentor. Furthermore, I would like to thank Prof. Dr. Burkhart Schraven for giving me the opportunity to work in his institute. His support, ideas, and the lively discussions promoted me to develop as a scientist. Special thanks go also to the whole AG Simeoni/Schraven - Ines, Camilla, Matthias, and Andreas - for the help with experiments, the discussions, and the fun we had. This helped to sustain also the longest days.
    [Show full text]
  • Mechanisms of Topical Bruton Tyrosine Kinase Inhibitor PRN473 P1572 in Immune‑Mediated Models of Skin Disease Yan Xing, Katherine A
    E‑Poster Mechanisms of Topical Bruton Tyrosine Kinase Inhibitor PRN473 P1572 in Immune‑Mediated Models of Skin Disease Yan Xing, Katherine A. Chu, Jyoti Wadhwa, Wei Chen, Jiang Zhu, Jin Shu, Matthew C. Foulke, Natalie Loewenstein, Philip Nunn, Kolbot By, Pasit Phiasivongsa, David M. Goldstein, and Claire L. Langrish Principia Biopharma Inc, A Sanofi Company, South San Francisco, CA, USA • In vitro, PRN473 inhibited B‑cell activation and FcR signaling in monocytes and mast cells, and IgE‑Mediated PCA Mouse Reaction INTRODUCTION MATERIALS AND METHODS in the absence of cytotoxic effects and limited functional effects in basophils, epidermal growth factor receptor (EGFR), and cells absent BTK signaling (Table 1A) • Significant dose‑dependent inhibition of mouse PCA IgE‑mediated reaction was also observed with single and multiday • PRN473 demonstrated durable BTK inhibition at 4 h that was partially sustained through 18 h applications of PRN473 ≥1%, approaching positive controls with higher strengths up to 4% PRN473 (Figure 6) Bruton Tyrosine Kinase In Vitro Selectivity and Functional Assays (Table 1B) • Multiday (3 days) treatment achieved similar levels of inhibition with respect to single administration (data not shown) • Bruton tyrosine kinase (BTK) is a promising target in immunology because of its expression in • PRN473 1 µM was tested against a panel of 230 kinase targets in the KINOMEscan™ panel B cells and innate immune cells, providing essential downstream signaling for B‑cell receptors, Biochemical studies were performed to characterize the potency, selectivity, biochemical Table 1. Preclinical Target Specificity and BTK Occupancy for PRN473 Figure 6. Dose‑Dependent Inhibition in IgE Antibody‑Mediated Mouse Passive Fc receptors, and other innate cell pathways (Figure 1)1,2 • on‑rate and off‑rate, and reversibility of the interaction between PRN473 and BTK Cutaneous Anaphylaxis (PCA) With Single Application of PRN473 • Selective inhibition of BTK function and occupancy and off‑target effects were evaluated in A.
    [Show full text]
  • Src-Family Kinases Impact Prognosis and Targeted Therapy in Flt3-ITD+ Acute Myeloid Leukemia
    Src-Family Kinases Impact Prognosis and Targeted Therapy in Flt3-ITD+ Acute Myeloid Leukemia Title Page by Ravi K. Patel Bachelor of Science, University of Minnesota, 2013 Submitted to the Graduate Faculty of School of Medicine in partial fulfillment of the requirements for the degree of Doctor of Philosophy University of Pittsburgh 2019 Commi ttee Membership Pa UNIVERSITY OF PITTSBURGH SCHOOL OF MEDICINE Commi ttee Membership Page This dissertation was presented by Ravi K. Patel It was defended on May 31, 2019 and approved by Qiming (Jane) Wang, Associate Professor Pharmacology and Chemical Biology Vaughn S. Cooper, Professor of Microbiology and Molecular Genetics Adrian Lee, Professor of Pharmacology and Chemical Biology Laura Stabile, Research Associate Professor of Pharmacology and Chemical Biology Thomas E. Smithgall, Dissertation Director, Professor and Chair of Microbiology and Molecular Genetics ii Copyright © by Ravi K. Patel 2019 iii Abstract Src-Family Kinases Play an Important Role in Flt3-ITD Acute Myeloid Leukemia Prognosis and Drug Efficacy Ravi K. Patel, PhD University of Pittsburgh, 2019 Abstract Acute myelogenous leukemia (AML) is a disease characterized by undifferentiated bone-marrow progenitor cells dominating the bone marrow. Currently the five-year survival rate for AML patients is 27.4 percent. Meanwhile the standard of care for most AML patients has not changed for nearly 50 years. We now know that AML is a genetically heterogeneous disease and therefore it is unlikely that all AML patients will respond to therapy the same way. Upregulation of protein-tyrosine kinase signaling pathways is one common feature of some AML tumors, offering opportunities for targeted therapy.
    [Show full text]
  • Current Understanding of Tyrosine Kinase BMX in Inflammation and Its
    Burns & Trauma, July 2014, Vol 2, Issue 3 Review Article Current understanding of tyrosine kinase BMX in infl ammation and its inhibitors Le Qiu, Fei Wang, Sheng Liu, Xu-Lin Chen Department of Burns, First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, China Corresponding author: Xu-Lin Chen, Department of Burns, First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui 230022, China. E-mail: [email protected] Received: 14-04-14, Revised: 05-06-14, Accepted: 11-06-14 ABSTRACT Tec family kinases, which include tyrosine kinase expressed in hepatocellular carcinoma (TEC), Bruton's tyrosine kinase (BTK), interleukin (IL)-2-inducible T-cell kinase (ITK), tyrosine-protein kinase (TXK), and bone marrow tyrosine kinase on chromosome X (BMX), are the second largest group of non-receptor tyrosine kinases and have a highly conserved carboxyl-terminal kinase domain. BMX was identifi ed in human bone marrow cells, and was demonstrated to have been expressed in myeloid hematopoietic lineages cells, endothelial cells, and several types of cancers. Signifi cant progress in this area during the last decade revealed an important role for BMX in infl ammation and oncologic disorders. This review focuses on BMX biology, its role in infl ammation and possible signaling pathways, and the potential of selective BMX inhibitors. Key words: BMX, infl ammation, tyrosine kinase Introduction tyrosine kinase on chromosome X (BMX).[2] Tec family kinases have been the focus of immunological interest ever The response of cells to extracellular stimuli is in part medi- since their discovery. ated by a number of intracellular kinases.
    [Show full text]
  • Review Bruton's Tyrosine Kinase Is Involved in Innate and Adaptive
    Histol Histopathol (2005) 20: 945-955 Histology and http://www.hh.um.es Histopathology Cellular and Molecular Biology Review Bruton’s Tyrosine Kinase is involved in innate and adaptive immunity C. Brunner, B. Müller and T. Wirth University of Ulm, Department of Physiological Chemistry, Ulm, Germany Summary. Btk is a cytoplasmic tyrosine kinase, which Btk is not restricted to B cells, it is also expressed in is mainly involved in B cell receptor signalling. Gene myeloid cells, like mast cells (Kawakami et al., 1994; targeting experiments revealed that Btk is important for Smith et al., 1994), monocytes/macrophages (de Weers B cell development and function. However, Btk is not et al., 1993; Smith et al., 1994), neutrophiles (Lachance only expressed in B cells, but also in most other et al., 2002), dendritic cells (DCs) (Gagliardi et al., haematopoietic lineages except for T cells and plasma 2003), in erythroid cells (Smith et al., 1994; Robinson et cells. Recently we found that Btk is involved in Toll-like al., 1998; Whyatt et al., 2000), in platelets (Quek et al., receptor signalling. Toll-like receptors play an important 1998), in hematopoietic stem cells (HSC), and role in innate immunity. They are highly expressed on multipotent progenitors (Phillips et al., 2000) as well as mast cells, macrophages and dendritic cells, which are in primary neuronal cells (Yang et al., 2004). Although essential for the recognition and consequently for the btk mutations cause primarily severe B cell defects, elimination of microbial pathogens. Therefore Btk might functions of btk-deficient or -mutated myeloid cells are play an important role for the function of also affected (see below).
    [Show full text]
  • The Bmx Tyrosine Kinase Is Activated by IL-3 and G-CSF in a PI-3K Dependent Manner
    Oncogene (2000) 19, 4151 ± 4158 ã 2000 Macmillan Publishers Ltd All rights reserved 0950 ± 9232/00 $15.00 www.nature.com/onc The Bmx tyrosine kinase is activated by IL-3 and G-CSF in a PI-3K dependent manner Niklas Ekman1,6, Elena Arighi1,2,6, Iiro Rajantie1, Pipsa Saharinen3, Ari RistimaÈ ki4, Olli Silvennoinen3,5 and Kari Alitalo*,1 1Molecular/Cancer Biology Laboratory and Ludwig Institute for Cancer Research, Haartman Institute, P.O.Box 21 (Haartmaninkatu 3), 00014 University of Helsinki, Finland; 2Department of Experimental Oncology, Istituto Nazionale Tumori, 20133 Milan, Italy; 3Department of Virology, Haartman Institute, P.O.Box 21 (Haartmaninkatu 3), 00014 University of Helsinki, Finland; 4Department of Obstetrics and Gynecology, Helsinki University Central Hospital, P.O. Box 140, 00029 Helsinki, Finland; 5Medical Biochemistry, University of Tampere, 33014 Tampere, Finland Cytoplasmic protein tyrosine kinases play crucial roles in The Tec family of PTKs consists of ®ve members: the signaling via a variety of cell surface receptors. The Bmx founder member Tec, as well as Btk, Itk/Tsk/Emt, Txk tyrosine kinase, a member of the Tec family, is and Bmx tyrosine kinases. The Tec kinases are regulated expressed in hematopoietic cells of the granulocytic and by a variety of hematopoietic receptors. For example, the monocytic lineages. Here we show that Bmx is Bruton's tyrosine kinase, Btk, is activated through the catalytically activated by interleukin-3 (IL-3) and high anity IgE receptor in mast cells, by the antigen granulocyte-colony stimulating factor (G-CSF) recep- and CD72 receptors in B-cells, as well as by interleukin-3 tors.
    [Show full text]
  • Protein Tyrosine Kinases: Their Roles and Their Targeting in Leukemia
    cancers Review Protein Tyrosine Kinases: Their Roles and Their Targeting in Leukemia Kalpana K. Bhanumathy 1,*, Amrutha Balagopal 1, Frederick S. Vizeacoumar 2 , Franco J. Vizeacoumar 1,3, Andrew Freywald 2 and Vincenzo Giambra 4,* 1 Division of Oncology, College of Medicine, University of Saskatchewan, Saskatoon, SK S7N 5E5, Canada; [email protected] (A.B.); [email protected] (F.J.V.) 2 Department of Pathology and Laboratory Medicine, College of Medicine, University of Saskatchewan, Saskatoon, SK S7N 5E5, Canada; [email protected] (F.S.V.); [email protected] (A.F.) 3 Cancer Research Department, Saskatchewan Cancer Agency, 107 Wiggins Road, Saskatoon, SK S7N 5E5, Canada 4 Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapies (ISBReMIT), Fondazione IRCCS Casa Sollievo della Sofferenza, 71013 San Giovanni Rotondo, FG, Italy * Correspondence: [email protected] (K.K.B.); [email protected] (V.G.); Tel.: +1-(306)-716-7456 (K.K.B.); +39-0882-416574 (V.G.) Simple Summary: Protein phosphorylation is a key regulatory mechanism that controls a wide variety of cellular responses. This process is catalysed by the members of the protein kinase su- perfamily that are classified into two main families based on their ability to phosphorylate either tyrosine or serine and threonine residues in their substrates. Massive research efforts have been invested in dissecting the functions of tyrosine kinases, revealing their importance in the initiation and progression of human malignancies. Based on these investigations, numerous tyrosine kinase inhibitors have been included in clinical protocols and proved to be effective in targeted therapies for various haematological malignancies.
    [Show full text]
  • Vascular Endothelial Growth Factor-B and The
    MARKUS RÄSÄNEN RÄSÄNEN MARKUS Recent Publications in this Series 38/2019 Ulrika Julku Prolyl Oligopeptidase and Alpha-Synuclein in the Regulation of Nigrostriatal Dopaminergic Neurotransmission 39/2019 Reijo Siren Screening for Cardiovascular Risk Factors in Middle-Aged Men: The Long-Term Effect of AND REVASCULARIZATION HYPERTROPHY IN CARDIAC KINASE AND THE BMX TYROSINE FACTOR-B GROWTH ENDOTHELIAL VASCULAR Lifestyle Counselling 40/2019 Paula Tiittala DISSERTATIONES SCHOLAE DOCTORALIS AD SANITATEM INVESTIGANDAM Hepatitis B and C, HIV and Syphilis among Migrants in Finland: Opportunities for Public UNIVERSITATIS HELSINKIENSIS Health Response 41/2019 Darshan Kumar Reticulon Homology Domain Containing Protein Families of the Endoplasmic Reticulum 42/2019 Iris Sevilem The Integration of Developmental Signals During Root Procambial Patterning in Arabidopsis thaliana 43/2019 Ying Liu MARKUS RÄSÄNEN Transcriptional Regulators Involved in Nutrient-Dependent Growth Control 44/2019 Ramón Pérez Tanoira VASCULAR ENDOTHELIAL GROWTH FACTOR-B AND Race for the Surface ― Competition Between Bacteria and Host Cells in Implant Colonization Process THE BMX TYROSINE KINASE IN CARDIAC HYPERTROPHY 45/2019 Mgbeahuruike Eunice Ego Evaluation of the Medicinal Uses and Antimicrobial Activity of Piper guineense (Schumach & AND REVASCULARIZATION Thonn) 46/2019 Suvi Koskinen Near-Occlusive Atherosclerotic Carotid Artery Disease: Study with Computed Tomography Angiography 47/2019 Flavia Fontana Biohybrid Cloaked Nanovaccines for Cancer Immunotherapy 48/2019 Marie Mennesson
    [Show full text]
  • BMX-ARHGAP Fusion Protein Maintains the Tumorigenicity Of
    Xu et al. Cancer Cell Int (2019) 19:133 https://doi.org/10.1186/s12935-019-0847-5 Cancer Cell International PRIMARY RESEARCH Open Access BMX-ARHGAP fusion protein maintains the tumorigenicity of gastric cancer stem cells by activating the JAK/STAT3 signaling pathway Xiao‑Feng Xu1†, Feng Gao2†, Jian‑Jiang Wang2, Cong Long1, Xing Chen2, Lan Tao3, Liu Yang1, Li Ding1 and Yong Ji2* Abstract Background: Cancer stem cells (CSCs), drug‑resistant cancer cell subsets, are known to be responsible for tumor metastasis and relapse. The JAK/STAT pathway, activated by SH2 domain, is known to regulate the tumor growth in gastric cancer (GC). Now, this study was designed to examine whether BMX‑ARHGAP afects the GC stem cell proper‑ ties and the underlying regulatory network via JAK/STAT axis. Methods: BMX‑ARHGAP expression was characterized in GC tissues and cells by RT‑qPCR and western blot assay. When BMX‑ARHGAP was overexpressed or silenced via plasmids or siRNA transfection, the stem cell properties were assessed by determining stem cell markers CD133, CD44, SOX2 and Nanog, followed by cell sphere and colony forma‑ tion assays. Subsequently, cell proliferation and invasion were examined by conducting EdU and Transwell assays. The JAK/STAT3 signaling pathway activation was inhibited using AG490. ARHGAP12, BMX exon 10–11, BXM‑SH2, JAK2 and STAT3 expression patterns were all determined to examine the regulatory network. The stem cell property in nude mice was also tested. Results: BMX‑ARHGAP was determined to be enriched in the GC. Overexpression of BMX‑ARHGAP resulted in increased expression of CD133, CD44, SOX2 and Nanog protein, and accelerated proliferation and invasion of CD133+CD44+ cells as well as facilitated self‑renewal potential of GC cells.
    [Show full text]
  • Datasheet Blank Template
    SAN TA C RUZ BI OTEC HNOL OG Y, INC . Bmx (C-17): sc-8874 BACKGROUND APPLICATIONS The Tec family of non-receptor tyrosine kinases is composed of six proteins, Bmx (C-17) is recommended for detection of Bmx of mouse, rat and human designated Tec, Emt (also known as Itk or Tsk), Btk (previously known as Atk, origin by Western Blotting (starting dilution 1:200, dilution range 1:100- BPK or Emb), Bmx, Txk (also known as Rlk) and Dsrc28C. All members of the 1:1000), immunoprecipitation [1-2 µg per 100-500 µg of total protein (1 ml family contain SH3 and SH2 domains and, with the exception of Txk and of cell lysate)], immunofluorescence (starting dilution 1:50, dilution range Dsrc28C, also contain a Pleckstrin homology (PH) and a Tec homology (TH) 1:50-1:500), immunohistochemistry (including paraffin-embedded sections) domain in their amino-termini. Four alternatively spliced forms of Tec are found (starting dilution 1:50, dilution range 1:50-1:500) and solid phase ELISA to be expressed broadly in cells of hematopoietic lineage and in hepatocytes. (starting dilution 1:30, dilution range 1:30-1:3000). The Emt gene product associates with CD28 and becomes activated subse - Bmx (C-17) is also recommended for detection of Bmx in additional species, quent to CD28 ligation. Btk is necessary for proper B cell development, and including equine, canine, bovine and porcine. mutations in the gene encoding Btk have been associated with families suf - fering from X-linked agammaglobulinemia, also referred to as Bruton’s disease.
    [Show full text]