P94fer Facilitates Cellular Recovery of Gamma Irradiated Pre-T Cells

Total Page:16

File Type:pdf, Size:1020Kb

P94fer Facilitates Cellular Recovery of Gamma Irradiated Pre-T Cells Oncogene (1997) 14, 2871 ± 2880 1997 Stockton Press All rights reserved 0950 ± 9232/97 $12.00 p94fer facilitates cellular recovery of gamma irradiated pre-T cells S Halachmy, O Bern, L Schreiber, M Carmel, Y Sharabi, J Shoham and U Nir Department of Life Sciences, Bar-Ilan University, Ramat-Gan 52900, Israel p94fer is a ubiquitous, nuclear and cytoplasmic tyrosine These three kinases encompass a group of the known, kinase, whose accumulation has been demonstrated in all SH2 containing mammalian nuclear tyrosine kinases mammalian cell lines analysed. In the present work, the (Yates et al., 1995; Wilks and Kurban, 1988; Van Etten p94fer expression pro®le was determined in cell lines et al., 1989; Hao et al., 1989, 1991) which are not src which were not tested before. While being present in related. While p94fer and c-Fes share similar structures, several hematopoietic and non hematopoietic cell lines containing a SH2 domain which is ¯anked by an including thymic stromal cells, the p94fer kinase could not extended N-terminal tail (Wilks and Kurban, 1988; be detected in pre-T and T cell lines. p94fer was also Hao et al., 1989), the c-Abl kinase contains both SH2 absent in pre-B line, but accumulated in these cells upon and SH3 domains and an extended C-terminal portion their induced development to antibody producing cells. (Ren et al., 1994). Several functional motifs have been This is in agreement with the absence of p94fer in primary de®ned in the Abl c-terminus, including a nuclear thymic and splenic T lymphocytes and its induced localization signal (NLS) (Van Etten et al., 1989), accumulation in stimulated B cells. Relatively high DNA binding domain (Kipreos and Wang, 1992), actin p94fer levels were detected in primary thymic and splenic binding domain (Van Etten et al., 1994; McWhirter stromal cells. Ectopic expression of p94fer in pre-T cells and Wang, 1993) and binding sites for the adapter slightly aected their cell cycle pro®le but it did not proteins Crk, Grb-2 and Nck (Ren et al., 1994). The aect their apoptotic death which was induced by N-terminal tail of p94fer exerts a coiled-coil structure ionizing radiation. However, p94fer facilitated dramati- which mediates the interaction of that kinase with cally, the cellular recovery of gamma irradiated pre-T other cellular proteins (Kim and Wang, 1995). Unlike cells which have escaped the apoptotic death. The c-Fes whose expression is con®ned to hematopoietic enhanced recovery of the irradiated, p94fer expressing progenitor cells (MacDonald et al., 1985; Care et al., pre-T cells, resulted most probably from their increased 1996), mature cells of the myeloid lineage (Hanazono survival, rather than from a prominent change in their et al., 1993; Greer et al., 1990; Feldman et al., 1985; proliferation rate. The absence of p94fer from pre-B and Care et al., 1996) and vascular endothelial cells (Greer pre-T cells, may thus contribute to the relative sensitivity et al., 1994), p94fer (Letwin et al., 1988; Hao et al., of these cells to ionizing radiation and to their 1989; Feldman et al., 1986) and c-Abl (Wang and dependence on the functioning of other nuclear tyrosine Baltimore, 1983; Van Etten et al., 1989; Renshaw et al., kinasese. 1988; Muller et al., 1982; Boulter and Wagner, 1988; Bernards et al., 1988) were reported to be ubiquitously Keywords: ionizing-radiation; nuclear; p94fer; T cell; expressed in all tissues analysed. Although diering in tyrosine kinase their tissue distribution, the subcellular localization of c-Fes and p94fer is similar and they accumulate both in the cytoplasm and nucleus of the expressing cells (Yates et al., 1995; Hao et al., 1991). While the cellular Introduction role of the c-Fes nuclear fraction is not understood, its cytoplasmic fraction can promote proliferation of p94fer is an evolutionary conserved (Pawson et al., vascular endothelial cells (Greer et al., 1990) and has 1989), nuclear and cytoplasmic tyrosine kinase (Hao et been implicated in cytokine signal transduction in al., 1991) whose presence has been documented in all hematopoietic cells (Izuhara et al., 1994; Hanazono et mammalian cell lines analysed (Letwin et al., 1988; al., 1993). The cytoplasmic fraction of p94fer interacts Hao et al., 1989; Feldman et al., 1986). A truncated with the catenin-like substrate pp120 in ®broblastic form of p94fer, termed p15ferT, accumulates solely in cells (Kim and Wang, 1995) and its nuclear fraction, meiotic primary spermatocytes (Hazan et al., 1993; was found to associate with the cell chromatin (Hao et Fischman et al., 1990) and is encoded by a testis al., 1991). The kinase activity of the cytoplasmic p94fer, speci®c FER transcript (Keshet et al., 1990; Fischman is elevated in growth factors stimulated ®broblastic cell et al., 1990). p52ferT and p94fer dier in their N-termini lines (Kim and Wang, 1995). However, no direct link but they do share a common SH2 and kinase domains of p94fer to regulation of cell growth has been shown, (Hao et al., 1989; Fischman et al., 1990). The p94fer nor has a p94fer substrate been characterized, thus kinase domain (Letwin et al., 1988; Hao et al., 1989) is leaving the cellular role of p94fer elusive. The 70% and 50% homologous to the kinase domains of ubiquitous expression of p94fer suggest that it may two other known nuclear tyrosine kinases c-Fes (Yates have a function that is fundamental to all cells. The et al., 1995; Wilks and Kurban, 1988; Roebroek et al., cellular levels of p94fer dier however among various 1985) and c-Abl (Van Etten et al., 1989), respectively. cell types (Feldman et al., 1986). While ®broblastic (Hao et al., 1989, 1991), epithelial (Hao et al., 1989, Correspondence: U Nir 1991) and macrophage (U Nir, unpublished data) cell Received 8 August 1996; revised 11 March 1997; accepted lines accumulate high p94fer levels, the expression of 11 March 1997 that tyrosine kinase in some promyelocytic cell lines is Cellular recovery of irradiated pre-T cells SHalachmyet al 2872 relatively low (Hao et al., 1989). In an attempt to et al., 1980). The 70Z/3 dierentiation system was thus elucidate the cellular functions of p94fer, the determina- adopted for checking the dependence of p94fer tion of its accumulation pro®le has been extended to accumulation, on the development of pre-B cells to cell types which have not been analysed before. In this antibody producing B cells. 70Z/3 cells were exposed to work we show that p94fer is not detected in pre-B, pre-T 10 mg/ml LPS and their p94fer accumulation pro®le was and T cell lines and that ectopic expression of p94fer in followed for up to 68 h of LPS treatment. Exposing the pre-T cells, dramatically improved their recovery after 70Z/3 pre-B cells to LPS treatment, led to gradual being exposed to ionizing radiation. The absence of accumulation of p94fer in the maturing cells (Figure 2). p94fer from pre-B and pre-T cells, may thus contribute The kinetic of p94fer accumulation lagged however, to their relative sensitivity to ionizing radiation and to behind the known kinetic of the k light chain the dependence of these cells on the functioning of production. While the k protein level reaches a other ubiquitous nuclear tyrosine kinases, like c-Abl (Tybulewicz et al., 1991; Schwarzberg et al., 1991). a S C2 R1 NIH TEC 127 B Results p94fer is not detected in pre-B, pre-T and T lymphocyte fer cell lines p94 ¨ To extend the previous analysis of the p94fer expression pro®le in various cell types (Letwin et al., 1988; Hao et al., 1989), the relative cellular levels of p94fer were determined in ®ve mouse cell lines which have not been 1234567 analysed before. These included two hematopoietic cell lines: R1 ± a friend erythroleukemia cell line (Marks et al., 1983) and cell line No 127 ± a mouse double positive pre-T thymoma (Irlin and Peled, 1992). The three non hematopoietic cell lines tested were: C2 ± a b 70Z TEC 127 T8 T4 NIH mouse myogenic cell line (Yae et al., 1996), STAC and TEC ± two thymic stromal epithelial cell lines p94fer ¨ (Glimcher et al., 1983). The p94fer levels in these cells were compared to those found in the previously analysed NIH3T3 mouse ®broblastic cell line and the human B cell line DAKIKI (Hao et al., 1989). Whole cell protein extracts prepared from the cultured cells, were fractionated in a SDS ± PAGE, Western blotted and the presence of p94fer was determined with the C1 p94fer antibodies. All the cell lines analysed except for 123456 127 (Figure 1a lane 6), were found to accumulate fer varying levels of the p94fer kinase. The absence of p94fer Figure 1 Accumulation of p94 in mammalian cell lines. (a) Whole cell proteins extracts from: lane 1, STAC cells; lane 2, C2 from the 127 pre-T cells, prompt us to check whether it myogenic cells; lane 3, R1 erythroleukemia cells; lane 4, NIH3T3; is expressed in pre-B and mature T cell lines. The lane 5, TEC cells; lane 6, pre-T 127 cells; and lane 7, DAKIKI B presence of p94fer was thus analysed in three other cells; were resolved in a 9% SDS ± PAGE, Western blotted and reacted with the C1 p94fer antibodies. Arrow on the left indicates mouse lymphocytic cell lines. These included the fer fer + the migration distance of p94 .(b) p94 accumulation in: lane 1, 2B4.11 cell line ± a CD4 T-cell hybridoma (Yang et 70Z/3 cells; lane 2, TEC cells; lane 3, 127 cells; lane 4, CTLL cells; al., 1993), CTLL ± a CD8+ IL-2 dependent cytotoxic T lane 5, 2B4.11 cells; lane 6, NIH3T3.
Recommended publications
  • Gene Symbol Gene Description ACVR1B Activin a Receptor, Type IB
    Table S1. Kinase clones included in human kinase cDNA library for yeast two-hybrid screening Gene Symbol Gene Description ACVR1B activin A receptor, type IB ADCK2 aarF domain containing kinase 2 ADCK4 aarF domain containing kinase 4 AGK multiple substrate lipid kinase;MULK AK1 adenylate kinase 1 AK3 adenylate kinase 3 like 1 AK3L1 adenylate kinase 3 ALDH18A1 aldehyde dehydrogenase 18 family, member A1;ALDH18A1 ALK anaplastic lymphoma kinase (Ki-1) ALPK1 alpha-kinase 1 ALPK2 alpha-kinase 2 AMHR2 anti-Mullerian hormone receptor, type II ARAF v-raf murine sarcoma 3611 viral oncogene homolog 1 ARSG arylsulfatase G;ARSG AURKB aurora kinase B AURKC aurora kinase C BCKDK branched chain alpha-ketoacid dehydrogenase kinase BMPR1A bone morphogenetic protein receptor, type IA BMPR2 bone morphogenetic protein receptor, type II (serine/threonine kinase) BRAF v-raf murine sarcoma viral oncogene homolog B1 BRD3 bromodomain containing 3 BRD4 bromodomain containing 4 BTK Bruton agammaglobulinemia tyrosine kinase BUB1 BUB1 budding uninhibited by benzimidazoles 1 homolog (yeast) BUB1B BUB1 budding uninhibited by benzimidazoles 1 homolog beta (yeast) C9orf98 chromosome 9 open reading frame 98;C9orf98 CABC1 chaperone, ABC1 activity of bc1 complex like (S. pombe) CALM1 calmodulin 1 (phosphorylase kinase, delta) CALM2 calmodulin 2 (phosphorylase kinase, delta) CALM3 calmodulin 3 (phosphorylase kinase, delta) CAMK1 calcium/calmodulin-dependent protein kinase I CAMK2A calcium/calmodulin-dependent protein kinase (CaM kinase) II alpha CAMK2B calcium/calmodulin-dependent
    [Show full text]
  • Cytokine Signaling Through the Novel Tyrosine Kinase RAFTK in Kaposi's
    Cytokine Signaling Through the Novel Tyrosine Kinase RAFTK in Kaposi’s Sarcoma Cells Zhong-Ying Liu,* Ramesh K. Ganju,*Jian-Feng Wang,* Mel A. Ona,* William C. Hatch,* Tong Zheng,‡ Shalom Avraham,* Parkash Gill,‡ and Jerome E. Groopman* *Divisions of Experimental Medicine and Hematology/Oncology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215; and ‡Division of Hematology/Oncology, Norris Cancer Center, University of Southern California, Los Angeles, California 90033 Abstract believed to be from the lymphatic endothelium (1–2). Etiolog- ical factors implicated in KS include the recently discovered A number of cytokines, including basic fibroblast growth human herpesvirus 8 (HHV-8)/Kaposi’s sarcoma herpesvirus factor (bFGF), vascular endothelial growth factor (VEGF), (KSHV) and TAT, the soluble transcriptional activator of oncostatin M (OSM), IL-6, and tumor necrosis factor alpha HIV (3–7). Considerable data indicate a role for endogenous (TNF-a), have been postulated to have a role in the patho- and exogenous cytokines in the pathogenesis of KS (8–16). genesis of Kaposi’s sarcoma (KS). The proliferative effects Growth factors such as basic fibroblast growth factor (bFGF) of bFGF and OSM may be via their reported activation of and vascular endothelial growth factor (VEGF), which are the c-Jun NH2-terminal kinase (JNK) signaling pathway in known to stimulate the mitogenesis of certain types of endo- KS cells. We now report that KS cells express a recently thelium, as well as Oncostatin M (OSM), IL-6, and tumor ne- identified focal adhesion kinase termed RAFTK which ap- crosis factor alpha (TNF-a) which are elaborated during in- pears in other cell systems to coordinate surface signals be- flammatory conditions, have been implicated in promoting KS tween cytokine and integrin receptors and the cytoskeleton cell growth (17–25).
    [Show full text]
  • Itk Tyrosine Kinase Substrate Docking Is Mediated by a Nonclassical SH2 Domain Surface of PLC␥1
    Itk tyrosine kinase substrate docking is mediated by a nonclassical SH2 domain surface of PLC␥1 Lie Min, Raji E. Joseph, D. Bruce Fulton, and Amy H. Andreotti1 Department of Biochemistry, Biophysics, and Molecular Biology, Iowa State University, Ames, IA 50011 Edited by Susan S. Taylor, University of California at San Diego, La Jolla, CA, and approved October 20, 2009 (received for review October 1, 2009) Interleukin-2 tyrosine kinase (Itk) is a Tec family tyrosine kinase that We have previously shown that the PLC␥1 SH2C domain mediates signaling processes after T cell receptor engagement. Acti- (spanning residues 659–756 within full-length PLC␥1) binds di- vation of Itk requires recruitment to the membrane via its pleckstrin rectly to the Itk kinase domain and is required for efficient homology domain, phosphorylation of Itk by the Src kinase, Lck, and phosphorylation of Y783 by Itk (26). Fragments of PLC␥1 that binding of Itk to the SLP-76/LAT adapter complex. After activation, Itk contain Y783 but not the SH2C domain are not efficiently phos- phosphorylates and activates phospholipase C-␥1 (PLC-␥1), leading to phorylated by Itk. Moreover, phosphorylation of PLC␥1 substrate production of two second messengers, DAG and IP3. We have previ- fragments that contain both SH2C and Y783 (spanning 659–789, ously shown that phosphorylation of PLC-␥1 by Itk requires a direct, hereafter referred to as PLC␥1 SH2C-linker) can be inhibited by phosphotyrosine-independent interaction between the Src homol- titration with isolated PLC␥1 SH2C domain (26). The excess, free ogy 2 (SH2) domain of PLC-␥1 and the kinase domain of Itk.
    [Show full text]
  • Supplementary Table 1. in Vitro Side Effect Profiling Study for LDN/OSU-0212320. Neurotransmitter Related Steroids
    Supplementary Table 1. In vitro side effect profiling study for LDN/OSU-0212320. Percent Inhibition Receptor 10 µM Neurotransmitter Related Adenosine, Non-selective 7.29% Adrenergic, Alpha 1, Non-selective 24.98% Adrenergic, Alpha 2, Non-selective 27.18% Adrenergic, Beta, Non-selective -20.94% Dopamine Transporter 8.69% Dopamine, D1 (h) 8.48% Dopamine, D2s (h) 4.06% GABA A, Agonist Site -16.15% GABA A, BDZ, alpha 1 site 12.73% GABA-B 13.60% Glutamate, AMPA Site (Ionotropic) 12.06% Glutamate, Kainate Site (Ionotropic) -1.03% Glutamate, NMDA Agonist Site (Ionotropic) 0.12% Glutamate, NMDA, Glycine (Stry-insens Site) 9.84% (Ionotropic) Glycine, Strychnine-sensitive 0.99% Histamine, H1 -5.54% Histamine, H2 16.54% Histamine, H3 4.80% Melatonin, Non-selective -5.54% Muscarinic, M1 (hr) -1.88% Muscarinic, M2 (h) 0.82% Muscarinic, Non-selective, Central 29.04% Muscarinic, Non-selective, Peripheral 0.29% Nicotinic, Neuronal (-BnTx insensitive) 7.85% Norepinephrine Transporter 2.87% Opioid, Non-selective -0.09% Opioid, Orphanin, ORL1 (h) 11.55% Serotonin Transporter -3.02% Serotonin, Non-selective 26.33% Sigma, Non-Selective 10.19% Steroids Estrogen 11.16% 1 Percent Inhibition Receptor 10 µM Testosterone (cytosolic) (h) 12.50% Ion Channels Calcium Channel, Type L (Dihydropyridine Site) 43.18% Calcium Channel, Type N 4.15% Potassium Channel, ATP-Sensitive -4.05% Potassium Channel, Ca2+ Act., VI 17.80% Potassium Channel, I(Kr) (hERG) (h) -6.44% Sodium, Site 2 -0.39% Second Messengers Nitric Oxide, NOS (Neuronal-Binding) -17.09% Prostaglandins Leukotriene,
    [Show full text]
  • FER Tyrosine Kinase (FER) Overexpression Mediates Resistance to Quinacrine Through EGF-Dependent Activation of NF-Κb
    FER tyrosine kinase (FER) overexpression mediates resistance to quinacrine through EGF-dependent activation of NF-κB Canhui Guo and George R. Stark1 Department of Molecular Genetics, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH 44195 Contributed by George R. Stark, April 5, 2011 (sent for review January 17, 2011) Quinacrine, a drug with antimalarial and anticancer activities that that overexpression of FER activates NF-κB, thus conferring inhibits NF-κB and activates p53, has progressed into phase II clin- resistance to the NF-κB inhibitor quinacrine. ical trials in cancer. To further elucidate its mechanism of action and identify pathways of drug resistance, we used an unbiased Results method for validation-based insertional mutagenesis to isolate Identification of FER in a Quinacrine-Resistant Clone. Eighteen dif- a quinacrine-resistant cell line in which an inserted CMV promoter ferent pools of human colon cancer RKO cells were infected with drives overexpression of the FER tyrosine kinase (FER). Overex- three different VBIM viruses (6) using a total of 1 million cells. pression of FER from a cDNA confers quinacrine resistance to sev- After propagation, each pool was replated and treated with 10 eral different types of cancer cell lines. We show that quinacrine μM quinacrine for 48 h. Twenty quinacrine-resistant colonies kills cancer cells primarily by inhibiting the activation of NF-κB and were observed 2 wk later in seven of the pools. The VBIM vectors that increased activation of NF-κB through FER overexpression contain LoxP sites, allowing excision of the promoter in candidate mediates resistance. EGF activates NF-κB and stimulates phosphor- mutant clones.
    [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; amb850@mail.usask.ca (A.B.); franco.vizeacoumar@usask.ca (F.J.V.) 2 Department of Pathology and Laboratory Medicine, College of Medicine, University of Saskatchewan, Saskatoon, SK S7N 5E5, Canada; frederick.vizeacoumar@usask.ca (F.S.V.); andrew.freywald@usask.ca (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: kak677@mail.usask.ca (K.K.B.); v.giambra@operapadrepio.it (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]
  • Mammary Gland Specific Expression of Brk/PTK6 Promotes Delayed
    Lofgren et al. Breast Cancer Research 2011, 13:R89 http://breast-cancer-research.com/content/13/5/R89 RESEARCHARTICLE Open Access Mammary gland specific expression of Brk/PTK6 promotes delayed involution and tumor formation associated with activation of p38 MAPK Kristopher A Lofgren1,2,3, Julie H Ostrander1,3, Daniel Housa1,3,4, Gregory K Hubbard1,3, Alessia Locatelli1,3, Robin L Bliss3, Kathryn L Schwertfeger2,3,5 and Carol A Lange1,2,3,6* Abstract Introduction: Protein tyrosine kinases (PTKs) are frequently overexpressed and/or activated in human malignancies, and regulate cancer cell proliferation, cellular survival, and migration. As such, they have become promising molecular targets for new therapies. The non-receptor PTK termed breast tumor kinase (Brk/PTK6) is overexpressed in approximately 86% of human breast tumors. The role of Brk in breast pathology is unclear. Methods: We expressed a WAP-driven Brk/PTK6 transgene in FVB/n mice, and analyzed mammary glands from wild-type (wt) and transgenic mice after forced weaning. Western blotting and immunohistochemistry (IHC) studies were conducted to visualize markers of mammary gland involution, cell proliferation and apoptosis, as well as Brk, STAT3, and activated p38 mitogen-activated protein kinase (MAPK) in mammary tissues and tumors from WAP-Brk mice. Human (HMEC) or mouse (HC11) mammary epithelial cells were stably or transiently transfected with Brk cDNA to assay p38 MAPK signaling and cell survival in suspension or in response to chemotherapeutic agents. Results: Brk-transgenic dams exhibited delayed mammary gland involution and aged mice developed infrequent tumors with reduced latency relative to wt mice.
    [Show full text]
  • Rare Genetic Variants Potentially Involved in Ovarian Hyperstimulation Syndrome
    Journal of Assisted Reproduction and Genetics (2019) 36:491–497 https://doi.org/10.1007/s10815-018-1372-5 GENETICS Rare genetic variants potentially involved in ovarian hyperstimulation syndrome Katrien Stouffs1 & Sari Daelemans2 & Samuel Santos-Ribeiro2 & Sara Seneca1 & Alexander Gheldof1 & Ali Sami Gürbüz3 & Michel De Vos2 & Herman Tournaye2 & Christophe Blockeel2 Received: 25 May 2018 /Accepted: 14 November 2018 /Published online: 27 November 2018 # Springer Science+Business Media, LLC, part of Springer Nature 2018 Abstract Purpose We aim to investigate whether there is a genetic predisposition in women who developed ovarian hyperstimulation syndrome (OHSS) after GnRH antagonist protocol with GnRH agonist trigger and freeze-all approach. Methods Four patients with OHSS after GnRH agonist trigger and freeze-all approach were gathered from the worldwide patient population. These patients were analyzed through Whole Exome Sequencing. In this study known causes of OHSS were investigated and new causes present in at least two individuals were searched for. Results In the first part of the study, we evaluated the presence of mutations in genes already known to be involved in OHSS. In PGR and TP53, heterozygous alterations were detected. PGR is predicted to be involved in progesterone resistance with a recessive inheritance pattern and is, therefore, not considered as being causal. The consequences of the variant detected in TP53 currently remain unknown. In part 2 of the study, we assessed the clinical significance of variants in genes previously not linked to OHSS. We especially focused on genes with variants present in ≥ 2 patients. Two patients have variants in the FLT4 gene. Mutations in this gene are linked to hereditary lymphedema, but no link to OHSS has been described.
    [Show full text]
  • Degradation of HER2/Neu by ANT2 Shrna Suppresses Migration and Invasiveness of Breast Cancer Cells Ji-Young Jang, Yoon-Kyung Jeon, Chul-Woo Kim*
    Jang et al. BMC Cancer 2010, 10:391 http://www.biomedcentral.com/1471-2407/10/391 RESEARCH ARTICLE Open Access Degradation of HER2/neu by ANT2 shRNA suppresses migration and invasiveness of breast cancer cells Ji-Young Jang, Yoon-Kyung Jeon, Chul-Woo Kim* Abstract Background: In breast cancer, the HER2/neu oncoprotein, which belongs to the epidermal growth factor receptor family, may trigger activation of the phosphoinositide-3 kinase (PI3K)/Akt pathway, which controls cell proliferation, survival, migration, and invasion. In this study, we examined the question of whether or not adenine nucleotide translocase 2 (ANT2) short hairpin RNA (shRNA)-mediated down-regulation of HER2/neu and inhibitory effects on the PI3K/Akt signaling pathway suppressed migration and invasiveness of breast cancer cells. Methods: We utilized an ANT2 vector-based RNA interference approach to inhibition of ANT2 expression, and the HER2/neu-overexpressing human breast cancer cell line, SK-BR3, was used throughout the study. Results: In this study, ANT2 shRNA decreased HER2/neu protein levels by promoting degradation of HER2/neu protein through dissociation from heat shock protein 90 (HSP90). As a result, ANT2 shRNA induced inhibitory effects on the PI3K/Akt signaling pathway. Inhibition of PI3K/Akt signaling by ANT2 shRNA caused down-regulation of membrane-type 1 matrix metalloproteinase (MT1-MMP) and vascular endothelial growth factor (VEGF) expression, decreased matrix metalloproteinase 2 (MMP2) and MMP9 activity, and suppressed migration and invasion of breast cancer cells. Conclusions: These results indicate that knock-down of ANT2 by shRNA down-regulates HER2/neu through suppression of HSP90’s function and inhibits the PI3K/Akt signaling pathway, resulting ultimately in suppressed migration and invasion of breast cancer cells.
    [Show full text]
  • Downloaded from Bioscientifica.Com at 09/23/2021 04:27:16PM Via Free Access Bernhard Et Al.: HER-2/Neu Vaccines
    Endocrine-Related Cancer (2002) 9 33–44 Vaccination against the HER-2/neu oncogenic protein H Bernhard1, L Salazar 2, K Schiffman2, A Smorlesi 2,3, B Schmidt1, K L Knutson2 andMLDisis2 1Technical University of Munich, Klinikum rechts der Isar, Department of Hematology and Oncology, Ismaningerstrasse 22, D-81664, Munich, Germany 2Division of Oncology, University of Washington, Seattle, Washington 98195–6527, USA 3Dipartimento Richerche INRCA, via Birarelli, 8, 60100 Ancona, Italy (Requests for offprints should be addressed to M L Disis, Box 356527, Oncology, University of Washington, Seattle, Washington 98195–6527, USA; Email: ndisis@u.washington.edu) Abstract The HER-2/neu oncogenic protein is a well-defined tumor antigen. HER-2/neu is a shared antigen among multiple tumor types. Patients with HER-2/neu protein-overexpressing breast, ovarian, non- small cell lung, colon, and prostate cancers have been shown to have a pre-existent immune response to HER-2/neu. No matter what the tumor type, endogenous immunity to HER-2/neu detected in cancer patients demonstrates two predominant characteristics. First, HER-2/neu-specific immune responses are found in only a minority of patients whose tumors overexpress HER-2/neu. Secondly, immunity, if detectable, is of low magnitude. These observations have led to the develop- ment of vaccine strategies designed to boost HER-2/neu immunity in a majority of patients. HER-2/ neu is a non-mutated self-protein, therefore vaccines must be developed based on immunologic principles focused on circumventing tolerance, a primary mechanism of tumor immune escape. HER-2/neu-specific vaccines have been tested in human clinical trials.
    [Show full text]
  • Downloads.Action)
    bioRxiv preprint doi: https://doi.org/10.1101/259192; this version posted February 2, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. INKA, an integrative data analysis pipeline for phosphoproteomic inference of active phosphokinases Thang V. Pham1,2, Robin Beekhof1,2, Carolien van Alphen1,2, Jaco C. Knol1, Alex A. Henneman1, Frank Rolfs1, Mariette Labots1, Evan Henneberry1, Tessa Y.S. Le Large1, Richard R. de Haas1, Sander R. Piersma1, Henk M.W. Verheul1, Connie R. Jimenez1 * 1 OncoProteomics Laboratory, Department of Medical Oncology, Cancer Center Amsterdam, VU University Medical Center, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands 2 These authors contributed equally to this work * Correspondence should be addressed to C.R.J. (c.jimenez@vumc.nl) Running Title: Integrative tool ranks active kinases Character Count: 50,356 1 bioRxiv preprint doi: https://doi.org/10.1101/259192; this version posted February 2, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 1 Abstract 2 3 Identifying (hyper)active kinases in cancer patient tumors is crucial to enable individualized 4 treatment with specific inhibitors. Conceptually, kinase activity can be gleaned from global 5 protein phosphorylation profiles obtained with mass spectrometry-based phosphoproteomics. A 6 major challenge is to relate such profiles to specific kinases to identify (hyper)active kinases that 7 may fuel growth/progression of individual tumors. Approaches have hitherto focused on 8 phosphorylation of either kinases or their substrates.
    [Show full text]
  • Supplementary Information Supplementary Table 1
    Supplementary information Supplementary Table 1 Kinase Type Format N Average SD (IC50) ALK Y Lantha 6 > 10 n.d. AURORA_A S/T Caliper 6 > 10 n.d. AXL Y Lantha 3 > 10 n.d. BTK Y Caliper 6 > 10 n.d. cABL Y Caliper 4 2.0 0.38 cABLT315 Y Caliper 6 > 10 n.d. CaMK2 S/T Caliper 3 > 10 n.d. CDK2A S/T Caliper 6 > 10 n.d. CDK4D1 S/T Caliper 3 > 10 n.d. CK1 S/T Caliper 3 > 10 n.d. cKIT Y Lantha 6 > 10 n.d. cMET Y Lantha 7 > 10 n.d. COT1 S/T Caliper 4 > 10 n.d. CSK Y Lantha 2 > 10 n.d. cSRC Y Lantha 3 > 10 n.d. EphA4 Y Lantha 6 > 10 n.d. EphB4 Y Lantha 6 > 10 n.d. ERK2 S/T Caliper 6 > 10 n.d. FAK Y Lantha 3 > 10 n.d. FGFR-1 Y Lantha 3 > 10 n.d. FGFR-2 Y Lantha 3 > 10 n.d. FGFR-3 Y Lantha 3 > 10 n.d. FGFR3K650E Y Lantha 6 > 10 n.d. FGFR-4 Y Lantha 6 > 10 n.d. FLT3 Y Lantha 3 > 10 n.d. FYN Y Lantha 3 > 10 n.d. GSK3beta S/T Caliper 6 > 10 n.d. HCK Y Lantha 3 > 10 n.d. HER1 Y Lantha 6 > 10 n.d. HER2 Y Lantha 6 > 10 n.d. HER4 Y Lantha 3 > 10 n.d. IGF1R Y Lantha 6 > 10 n.d. INS1R Y Lantha 6 > 10 n.d. IRAK4 S/T Caliper 3 > 10 n.d.
    [Show full text]