DOCSLIB.ORG

Conserved Features of Cancer Cells Define Their Sensitivity to HAMLET

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Conserved Features of Cancer Cells Define Their Sensitivity to HAMLET Oncogene (2011) 30, 4765–4779 & 2011 Macmillan Publishers Limited All rights reserved 0950-9232/11 www.nature.com/onc ORIGINAL ARTICLE Conserved features of cancer cells define their sensitivity to HAMLET-induced death; c-Myc and glycolysis P Storm1,5, S Aits1,5, MK Puthia2,5, A Urbano2, T Northen3, S Powers4, B Bowen3, YChao2, W Reindl3, DY Lee3, NL Sullivan4, J Zhang4, M Trulsson1, H Yang2, JD Watson4 and C Svanborg1 1Division of Microbiology, Immunology and Glycobiology, Department of Laboratory Medicine, Lund University, Lund, Sweden; 2Singapore Immunology Network (SIgN), Biomedical Sciences Institutes, Agency for Science, Technology, and Research (A*STAR), Singapore; 3Lawrence Berkeley National Laboratory, Berkeley, CA, USA and 4Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, USA HAMLET is the first member of a new family of Introduction tumoricidal protein–lipid complexes that kill cancer cells broadly, while sparing healthy, differentiated cells. Many Despite the molecular complexity of oncogenic trans- and diverse tumor cell types are sensitive to the lethal formation, cancer cells frequently suffer from ‘oncogene effect, suggesting that HAMLET identifies and activates addiction’, and the disruption of a single oncogene can conserved death pathways in cancer cells. Here, we either reverse oncogenesis or be lethal (Weinstein and investigated the molecular basis for the difference in Joe, 2008). The c-Myc oncogene is a classic example sensitivity between cancer cells and healthy cells. Using a (Felsher and Bishop, 1999) and is deregulated in at least combination of small-hairpin RNA (shRNA) inhibition, 40% of all human cancers (Dang et al., 2009). The proteomic and metabolomic technology, we identified the c- broad transforming effect of c-Myc has been explained Myc oncogene as one essential determinant of HAMLET by its ability to bind to promoters of at least 30% of all sensitivity. Increased c-Myc expression levels promoted known genes (Dang et al., 2009), and in transgenic mice, sensitivity to HAMLET and shRNA knockdown of c-Myc c-Myc overexpression combined with inhibition of suppressed the lethal response, suggesting that oncogenic apoptosis is sufficient to drive pancreatic cancer forma- transformation with c-Myc creates a HAMLET-sensitive tion (Pelengaris et al., 2002). More recently, inhibition phenotype. Furthermore, HAMLET sensitivity was mod- of c-Myc was proposed to stop cancer growth and even ified by the glycolytic state of tumor cells. Glucose allow tissue repair and reversion to a functional deprivation sensitized tumor cells to HAMLET-induced phenotype (Soucek et al., 2008). Healthy cells, in cell death and in the shRNA screen, hexokinase 1 (HK1), contrast, retain a limited replicative potential, due to 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 1 and their susceptibility to programmed cell death, anti- hypoxia-inducible factor 1a modified HAMLET sensitiv- growth signals and other molecular interactions that ity. HK1 was shown to bind HAMLET in a protein array limit longevity and maintain normal tissue homeostasis containing B8000 targets, and HK activity decreased (Hanahan and Weinberg, 2000, 2011). within 15 min of HAMLET treatment, before morpholo- Oncogene activity may also cause a shift in the gical signs of tumor cell death. In parallel, HAMLET glycolytic machinery (Hsu and Sabatini, 2008), and triggered rapid metabolic paralysis in carcinoma cells. cancer cells are highly dependent on glycolysis (Mathu- Tumor cells were also shown to contain large amounts of pala et al., 1997). Glucose is first trapped inside cells by oleic acid and its derivatives already after 15 min. The hexokinases 1 and 2 (HK1 and HK2) (Wilson, 2003) and results identify HAMLET as a novel anti-cancer agent that then further converted to pyruvate. In contrast to killstumorcellsbyexploitingunifyingfeaturesofcancer healthy cells, cancer cells drive pyruvate to lactate cells such as oncogene addiction or the Warburg effect. conversion even in the presence of oxygen, a process Oncogene (2011) 30, 4765–4779; doi:10.1038/onc.2011.196; known as the Warburg effect (Warburg, 1956). Recent published online 6 June 2011 studies have shown that oxidative phosphorylation is functional in most cancer cells (Vander Heiden et al., Keywords: HAMLET; metabolism; c-Myc; glycolysis 2009) and have proposed that the Warburg effect may reflect the expression of the pyruvate kinase isoform M2 (PKM2) (Christofk et al., 2008), which catalyzes the dephosphorylation of phosphoenolpyruvate to pyruvate Correspondence: Professor C Svanborg, Department of Microbiology, and is responsible for net ATP production within the Immunology and Glycobiology (MIG), Institute of Laboratory glycolytic sequence. In contrast to healthy differentiated Medicine, Lund University, So¨lvegatan 23, S-223 62 Lund, Sweden. cells, embryonic cells and most cancer cells mainly E-mail: [email protected] express the M2 isoform (Mazurek et al., 2005), and in 5Shared first author. Received 22 December 2010; revised and accepted 19 April 2011; some tumors, replacement of M2 by M1 reverses the published online 6 June 2011 Warburg effect and reduces tumorigenicity (Christofk Metabolism and c-Myc defines HAMLET sensitivity P Storm et al 4766 et al., 2008). In addition to PKM2, c-Myc has a A small number of shRNA vectors were enriched or fundamental role in reprogramming metabolism in depleted in the population treated with HAMLET; 43 certain tumor cells and has been shown to enhance inhibitory and 44 sensitizing shRNAs showed an aerobic glycolysis by directly activating genes, such as absolute MaxMean score of 40.5 in both high- and HK2, PKM1 and LDHA (Shim et al., 1997; Dang et al., low-dose treatment groups or an absolute score of 42in 2009). The transcription factor hypoxia-inducible factor any group (Figure 1b and Supplementary Table S1). The 1 (HIF1) also regulates glycolysis by enhancing the identified genes included oncogenes like c-Myc and expression of glucose transporters, HKs and phospho- glycolysis-related proteins. shRNAs against HK1 and fructokinases (PFKs) (Denko, 2008). HIF1a sensitized the cells to HAMLET (Figures 1c and HAMLET is a complex of partially unfolded a- d), suggesting that increased HK1 and HIF1a expres- lactalbumin and oleic acid, which preferentially kills sion drives a phenotype more resistant to HAMLET. In cancer cells and immature cells (Svanborg et al., 2003). contrast, shRNA targeting 6-phosphofructo-2-kinase/ The broad tumoricidal activity includes leukemia cells, fructose-2,6-biphosphatase (PFKFB1) and c-Myc res- carcinoma cells and glioma cells, whereas healthy, cued the cells (Figures 1c and d). Biological pathways differentiated cells survive HAMLET challenge (Fischer enriched for by HAMLET were identified by gene set et al., 2004). HAMLET thus appears to target highly enrichment analysis (GSEA) using the canonical path- conserved survival mechanisms that are crucial for ways gene set collection. The GSEA algorithm tests for cancer cells. This selectivity has been confirmed in vivo the enrichment of a set of items within a larger ranked in a human glioblastoma xenograft model and in a list of such items. The identified pathways confirmed the murine bladder cancer model (Fischer et al., 2004; importance of glycolysis, as four glycolysis-related Mossberg et al., 2010). In clinical studies of skin pathways significantly modified HAMLET sensitivity papillomas and bladder cancers (Gustafsson et al., (Figures 1e and f). In addition, the Ras pathway, which 2004; Mossberg et al., 2007) HAMLET showed governs c-Myc expression, was identified as a determi- tumoricidal activity with limited side effects. nant of HAMLET sensitivity (Figure 1f). This study investigated the molecular basis of HAM- The shRNA screen suggested that oncogenic trans- LET sensitivity in cancer cells, using screening techni- formation by c-Myc sensitizes cells to HAMLET- ques to detect features of transcription, protein induced cell death and that the glycolytic machinery interaction and metabolism that alter survival after influences HAMLET sensitivity of cancer cells. HAMLET exposure. We show that sensitivity to HAMLET is dependent on c-Myc and glycolysis and identified the glycolytic enzyme HK1 as a potential The c-Myc oncogene determines the tumoricidal effect direct target of HAMLET. The results also indicated of HAMLET that HAMLET alters cancer cell metabolism and that it We subsequently examined whether HAMLET sensitiv- may disrupt glycolysis, which is altered in cancer cells ity reflects c-Myc expression levels. Human lung due to the Warburg effect. carcinoma cells (A549), kidney carcinoma cells (A498) and semi-differentiated healthy kidney cells (human renal proximal tubule cells) were exposed to HAMLET Results (21 mM), and ATP levels monitored (Figure 2a). A rapid reduction in ATP levels was observed in lung and kidney An shRNA screen identifies c-Myc and glycolytic-related carcinoma cells but healthy cells remained unaffected proteins as modifiers of HAMLET sensitivity after 3, 6 and 24 h. However, at higher HAMLET To get an unbiased overview of genes important for concentrations, a partial response was also seen in those HAMLET sensitivity, we used the RNA interference cells (Supplementary Figure S1A). The difference in c- barcode screening technology (Brummelkamp and Myc expression reflected the difference in HAMLET Bernards, 2003; Silva et al., 2005, 2008). Barcode sensitivity, as lung and kidney carcinoma cells show screens use DNA microarrays to follow the relative
Load more

Recommended publications
  • Deep Learning–Based Multi-Omics Integration Robustly Predicts Survival in Liver Cancer Kumardeep Chaudhary1, Olivier B
    Published OnlineFirst October 5, 2017; DOI: 10.1158/1078-0432.CCR-17-0853 Statistics in CCR Clinical Cancer Research Deep Learning–Based Multi-Omics Integration Robustly Predicts Survival in Liver Cancer Kumardeep Chaudhary1, Olivier B. Poirion1, Liangqun Lu1,2, and Lana X. Garmire1,2 Abstract Identifying robust survival subgroups of hepatocellular car- index (C-index) ¼ 0.68]. More aggressive subtype is associated cinoma (HCC) will significantly improve patient care. Current- with frequent TP53 inactivation mutations, higher expression ly, endeavor of integrating multi-omicsdatatoexplicitlypredict of stemness markers (KRT19 and EPCAM)andtumormarker HCC survival from multiple patient cohorts is lacking. To fill BIRC5, and activated Wnt and Akt signaling pathways. We this gap, we present a deep learning (DL)–based model on HCC validated this multi-omics model on five external datasets of that robustly differentiates survival subpopulations of patients various omics types: LIRI-JP cohort (n ¼ 230, C-index ¼ 0.75), in six cohorts. We built the DL-based, survival-sensitive model NCI cohort (n ¼ 221, C-index ¼ 0.67), Chinese cohort (n ¼ on 360 HCC patients' data using RNA sequencing (RNA-Seq), 166, C-index ¼ 0.69), E-TABM-36 cohort (n ¼ 40, C-index ¼ miRNA sequencing (miRNA-Seq), and methylation data from 0.77), and Hawaiian cohort (n ¼ 27, C-index ¼ 0.82). This TheCancerGenomeAtlas(TCGA),whichpredictsprognosis is the first study to employ DL to identify multi-omics features as good as an alternative model where genomics and clinical linked to the differential survival of patients with HCC. Given data are both considered. This DL-based model provides two its robustness over multiple cohorts, we expect this workflow to optimal subgroups of patients with significant survival differ- be useful at predicting HCC prognosis prediction.
    [Show full text]
  • Small-Molecule Inhibition of 6-Phosphofructo-2-Kinase Activity Suppresses Glycolytic Flux and Tumor Growth
    110 Small-molecule inhibition of 6-phosphofructo-2-kinase activity suppresses glycolytic flux and tumor growth Brian Clem,1,3 Sucheta Telang,1,3 Amy Clem,1,3 reduces the intracellular concentration of Fru-2,6-BP, Abdullah Yalcin,1,2,3 Jason Meier,2 glucose uptake, and growth of established tumors in vivo. Alan Simmons,1,3 Mary Ann Rasku,1,3 Taken together, these data support the clinical development Sengodagounder Arumugam,1,3 of 3PO and other PFKFB3 inhibitors as chemotherapeutic William L. Dean,2,3 John Eaton,1,3 Andrew Lane,1,3 agents. [Mol Cancer Ther 2008;7(1):110–20] John O. Trent,1,2,3 and Jason Chesney1,2,3 Departments of 1Medicine and 2Biochemistry and Molecular Introduction Biology and 3Molecular Targets Group, James Graham Brown Neoplastic transformation causes a marked increase in Cancer Center, University of Louisville, Louisville, Kentucky glucose uptake and catabolic conversion to lactate, which forms the basis for the most specific cancer diagnostic 18 Abstract examination—positron emission tomography of 2- F- fluoro-2-deoxyglucose (18F-2-DG) uptake (1). The protein 6-Phosphofructo-1-kinase, a rate-limiting enzyme of products of several oncogenes directly increase glycolytic glycolysis, is activated in neoplastic cells by fructose-2,6- flux even under normoxic conditions, a phenomenon bisphosphate (Fru-2,6-BP), a product of four 6-phospho- originally termed the Warburg effect (2, 3). For example, fructo-2-kinase/fructose-2,6-bisphosphatase isozymes c-myc is a transcription factor that promotes the expression (PFKFB1-4). The inducible PFKFB3 isozyme is constitu- of glycolytic enzyme mRNAs, and its expression is increased tively expressed by neoplastic cells and required for the in several human cancers regardless of the oxygen pressure high glycolytic rate and anchorage-independent growth of (4, 5).
    [Show full text]
  • Exploring the Non-Canonical Functions of Metabolic Enzymes Peiwei Huangyang1,2 and M
    © 2018. Published by The Company of Biologists Ltd | Disease Models & Mechanisms (2018) 11, dmm033365. doi:10.1242/dmm.033365 REVIEW SPECIAL COLLECTION: CANCER METABOLISM Hidden features: exploring the non-canonical functions of metabolic enzymes Peiwei Huangyang1,2 and M. Celeste Simon1,3,* ABSTRACT A key finding from studies of metabolic enzymes is the existence The study of cellular metabolism has been rigorously revisited over the of mechanistic links between their nuclear localization and the past decade, especially in the field of cancer research, revealing new regulation of transcription. By modulating gene expression, insights that expand our understanding of malignancy. Among these metabolic enzymes themselves facilitate adaptation to rapidly insights isthe discovery that various metabolic enzymes have surprising changing environments. Furthermore, they can directly shape a ’ activities outside of their established metabolic roles, including in cell s epigenetic landscape (Kaelin and McKnight, 2013). the regulation of gene expression, DNA damage repair, cell cycle Strikingly, several metabolic enzymes exert completely distinct progression and apoptosis. Many of these newly identified functions are functions in different cellular compartments. Nuclear fructose activated in response to growth factor signaling, nutrient and oxygen bisphosphate aldolase, for example, directly interacts with RNA ́ availability, and external stress. As such, multifaceted enzymes directly polymerase III to control transcription (Ciesla et al., 2014),
    [Show full text]
  • Adiponectin Regulates Expression of Hepatic Genes Critical for Glucose and Lipid Metabolism
    Adiponectin regulates expression of hepatic genes critical for glucose and lipid metabolism Qingqing Liua, Bingbing Yuana, Kinyui Alice Loa,b, Heide Christine Pattersona,c,d, Yutong Sune, and Harvey F. Lodisha,b,1 aWhitehead Institute for Biomedical Research, Cambridge, MA 02142; bDepartment of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139; cDepartment of Pathology, Brigham and Women’s Hospital, Boston, MA 02115; dInstitut für Klinische Chemie und Pathobiochemie, Klinikum Rechts der Isar, Technische Universität München, 81675 Munich, Germany; and eDepartment of Molecular and Cellular Oncology, University of Texas MD Anderson Cancer Center, Houston, TX 77030 Contributed by Harvey F. Lodish, July 23, 2012 (sent for review May 30, 2012) The effects of adiponectin on hepatic glucose and lipid metabolism mediated by a master regulator of hepatic gene expression, the at transcriptional level are largely unknown. We profiled hepatic transcription factor (TF) Hnf4a. gene expression in adiponectin knockout (KO) and wild-type (WT) mice by RNA sequencing. Compared with WT mice, adiponectin KO Results mice fed a chow diet exhibited decreased mRNA expression of rate- Rate-limiting enzymes catalyze the slowest or irreversible steps limiting enzymes in several important glucose and lipid metabolic in metabolic pathways, which makes them the targets of tran- pathways, including glycolysis, tricarboxylic acid cycle, fatty-acid scriptional regulation and/or posttranslational modifications activation and synthesis, triglyceride synthesis, and cholesterol such as phosphorylation in the regulation of metabolism (7). synthesis. In addition, binding of the transcription factor Hnf4a to Because adiponectin plays an important role in energy metabo- DNAs encoding several key metabolic enzymes was reduced in KO lism, our analysis on hepatic gene expression in adiponectin KO mice, suggesting that adiponectin might regulate hepatic gene and WT mice was specifically focused on the expression levels of expression via Hnf4a.
    [Show full text]
  • Histone Deacetylase Inhibitors
    ANTICANCER RESEARCH 36 : 5019-5024 (2016) doi:10.21873/anticanres.11070 Review Histone Deacetylase Inhibitors: A Novel Therapeutic Weapon Αgainst Medullary Thyroid Cancer? CHRISTOS DAMASKOS 1,2* , SERENA VALSAMI 3* , ELEFTHERIOS SPARTALIS 2, EFSTATHIOS A. ANTONIOU 1, PERIKLIS TOMOS 4, STEFANOS KARAMAROUDIS 5, THEOFANO ZOUMPOU 5, VASILIOS PERGIALIOTIS 2, KONSTANTINOS STERGIOS 2,6 , CONSTANTINOS MICHAELIDES 7, KONSTANTINOS KONTZOGLOU 1, DESPINA PERREA 2, NIKOLAOS NIKITEAS 2 and DIMITRIOS DIMITROULIS 1 1Second Department of Propedeutic Surgery, “Laiko” General Hospital, Medical School, National and Kapodistrian University of Athens, Athens, Greece; 2Laboratory of Experimental Surgery and Surgical Research N.S. Christeas, Medical School, National and Kapodistrian University of Athens, Athens, Greece; 3Blood Transfusion Department, Aretaieion Hospital, Medical School, National and Kapodistrian Athens University, Athens, Greece; 4Department of Thoracic Surgery, “Attikon” General Hospital, Medical School, National and Kapodistrian University of Athens, Athens, Greece; 5Medical School, National and Kapodistrian University of Athens, Athens, Greece; 6Colorectal Department, General Surgery, The Princess Alexandra Hospital NHS Trust, Harlow, U.K.; 71st Department of Pathology, School of Medicine, University of Athens, Athens, Greece Abstract. Background/ Aim : Medullary thyroid cancer (MTC) is and histone deacetylase (HDAC) seems to play a potential role highly malignant, metastatic and recurrent, remaining generally to gene transcription. On the
    [Show full text]
  • Glycolytic Reliance Promotes Anabolism in Photoreceptors
    bioRxiv preprint doi: https://doi.org/10.1101/101964; this version posted January 21, 2017. 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. Glycolytic reliance promotes anabolism in photoreceptors Yashodhan Chinchore, Tedi Begaj, David Wu, Eugene Drokhlyansky, Constance L. Cepko* *Departments of Genetics and Ophthalmology, Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts 02115, USA [email protected] bioRxiv preprint doi: https://doi.org/10.1101/101964; this version posted January 21, 2017. 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 Sensory neurons capture information from the environment and convert it 2 into signals that can greatly impact the survival of an organism. These systems 3 are thus under heavy selective pressure, including for the most efficient use of 4 energy to support their sensitivity and efficiency1. In this regard, the 5 vertebrate photoreceptor cells face a dual challenge. They not only need to 6 preserve their membrane excitability via ion pumps by ATP hydrolysis2 but 7 also maintain a highly membrane rich organelle, the outer segment, which is 8 the primary site of phototransduction, creating a considerable biosynthetic 9 demand. How photoreceptors manage carbon allocation to balance their 10 catabolic and anabolic demands is poorly understood. One metabolic feature 11 of the retina is its ability to convert the majority of its glucose into lactate3,4 12 even in the presence of oxygen.
    [Show full text]
  • Metabolic Regulation of Calcium Pumps in Pancreatic Cancer: Role of Phosphofructokinase-Fructose- Bisphosphatase-3 (PFKFB3) D
    Richardson et al. Cancer & Metabolism (2020) 8:2 https://doi.org/10.1186/s40170-020-0210-2 RESEARCH Open Access Metabolic regulation of calcium pumps in pancreatic cancer: role of phosphofructokinase-fructose- bisphosphatase-3 (PFKFB3) D. A. Richardson1, P. Sritangos1, A. D. James2, A. Sultan1 and J. I. E. Bruce1* Abstract Background: High glycolytic rate is a hallmark of cancer (Warburg effect). Glycolytic ATP is required for fuelling plasma membrane calcium ATPases (PMCAs), responsible for extrusion of cytosolic calcium, in pancreatic ductal adenocarcinoma (PDAC). Phosphofructokinase-fructose-bisphosphatase-3 (PFKFB3) is a glycolytic driver that activates key rate-limiting enzyme Phosphofructokinase-1; we investigated whether PFKFB3 is required for PMCA function in PDAC cells. Methods: PDAC cell-lines, MIA PaCa-2, BxPC-3, PANC1 and non-cancerous human pancreatic stellate cells (HPSCs) were used. Cell growth, death and metabolism were assessed using sulforhodamine-B/tetrazolium-based assays, poly-ADP- ribose-polymerase (PARP1) cleavage and seahorse XF analysis, respectively. ATP was measured using a luciferase-based assay, membrane proteins were isolated using a kit and intracellular calcium concentration and PMCA activity were measured using Fura-2 fluorescence imaging. Results: PFKFB3 was highly expressed in PDAC cells but not HPSCs. In MIA PaCa-2, a pool of PFKFB3 was identified at the plasma membrane. PFKFB3 inhibitor, PFK15, caused reduced cell growth and PMCA activity, leading to calcium overload and apoptosis in PDAC cells. PFK15 reduced glycolysis but had noeffectonsteady-stateATPconcentrationinMIAPaCa-2. Conclusions: PFKFB3 is important for maintaining PMCA function in PDAC, independently of cytosolic ATP levels and may be involved in providing a localised ATP supply at the plasma membrane.
    [Show full text]
  • Protein–Lipid Complexes: Molecular Structure, Current Scenarios and Mechanisms of Cytotoxicity Cite This: RSC Adv.,2019,9, 36890 Esmail M
    RSC Advances REVIEW View Article Online View Journal | View Issue Protein–lipid complexes: molecular structure, current scenarios and mechanisms of cytotoxicity Cite this: RSC Adv.,2019,9, 36890 Esmail M. El-Fakharany *a and Elrashdy M. Redwan ab Some natural proteins can be complexed with oleic acid (OA) to form an active protein–lipid formulation that can induce tumor-selective apoptosis. The first explored protein was human milk a-lactalbumin (a- LA), called HAMLET when composed with OA in antitumor form. Several groups have prepared active protein–lipid complexes using a variety of approaches, all of which depend on target protein destabilization or direct OA–protein incubation to alter pH to acid or alkaline condition. In addition to performing vital roles in inflammatory processes and immune responses, fatty acids can disturb different metabolic pathways and cellular signals. Therefore, the tumoricidal action of these complexes is related to OA rather than the protein that keeps OA in solution and acts as a vehicle for transferring OA molecules to tumor cells. However, other studies have suggested that the antitumor efficacy of these Received 5th September 2019 complexes was exerted by both protein and OA together. The potential is not limited to the anti-tumor Creative Commons Attribution 3.0 Unported Licence. Accepted 21st October 2019 activity of protein–lipid complexes but extends to other functions such as bactericidal activity. The DOI: 10.1039/c9ra07127j protein shell enhances the solubility and stability of the bound fatty acid. These protein–lipid complexes rsc.li/rsc-advances are promising candidates for fighting various cancer types and managing bacterial and viral infections.
    [Show full text]
  • Cyclin and Growth Arrest-Specific Genes
    ISSN: 0975-8585 Research Journal of Pharmaceutical, Biological and Chemical Sciences Expression of Genes Encoded 6-phosphofructo-2-kinase/fructose-2, 6- bisphosphatase and 6-phosphofructo-1-kinase in U87 Glioma Cells with ERN1 Loss of Function: Hypoxic Regulation 1,2Minchenko DM, 1Lypova NM, 1Harmash YA, 3Kulinich AO, 1Marunych RY, 1Karbovskyi LL, 1*Minchenko ОH 1Palladin Institute of Biochemistry National Academy of Science of Ukraine, Kyiv, Ukraine. 2National O.O. Bohomolets Medical University, Kyiv 01601, Ukraine. 3Dnipropetrovs’k State Medical Academy, Dnipropetrovs’k 49600, Ukraine. ABSTRACT The endoplasmic reticulum–nuclei-1 (ERN1) sensing and signaling enzyme mediates a set of complex intracellular signaling events known as the unfolded protein response. We have studied the effect of hypoxia on the expression of genes encoded different 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFKFB1, PFKFB2, PFKFB3 and PFKFB4) and 6-phosphofructo-1-kinase (PFKL, PFKM and PFKP) as well as lactate dehydrogenase (LDHA) in glioma U87 cells and its subline with suppressed function of ERN1 signaling enzyme. It was shown that blockade a function of ERN1 enzyme, the key endoplasmic reticulum stress sensor, leads to an increase in the expression levels of PFKFB1, PFKFB2, PFKFB3 and PFKFB4 mRNA, being more significant for PFKFB4 and PFKFB2. Moreover, the expression level of PFKL and PFKP as well as LDHA mRNA also increases in cells with ERN1 loss of function, being more significant for PFKL. At the same time, the expression level of PFKM mRNA significantly decreases at this experimental condition. Exposure cells under hypoxic conditions leads to an increase of the expression level of PFKFB3 and PFKFB4 mRNA both in control glioma cells and cells with ERN1 loss of function, being more significant for PFKFB4.
    [Show full text]
  • A Graph-Theoretic Approach to Model Genomic Data and Identify Biological Modules Asscociated with Cancer Outcomes
    A Graph-Theoretic Approach to Model Genomic Data and Identify Biological Modules Asscociated with Cancer Outcomes Deanna Petrochilos A dissertation presented in partial fulfillment of the requirements for the degree of Doctor of Philosophy University of Washington 2013 Reading Committee: Neil Abernethy, Chair John Gennari, Ali Shojaie Program Authorized to Offer Degree: Biomedical Informatics and Health Education UMI Number: 3588836 All rights reserved INFORMATION TO ALL USERS The quality of this reproduction is dependent upon the quality of the copy submitted. In the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if material had to be removed, a note will indicate the deletion. UMI 3588836 Published by ProQuest LLC (2013). Copyright in the Dissertation held by the Author. Microform Edition © ProQuest LLC. All rights reserved. This work is protected against unauthorized copying under Title 17, United States Code ProQuest LLC. 789 East Eisenhower Parkway P.O. Box 1346 Ann Arbor, MI 48106 - 1346 ©Copyright 2013 Deanna Petrochilos University of Washington Abstract Using Graph-Based Methods to Integrate and Analyze Cancer Genomic Data Deanna Petrochilos Chair of the Supervisory Committee: Assistant Professor Neil Abernethy Biomedical Informatics and Health Education Studies of the genetic basis of complex disease present statistical and methodological challenges in the discovery of reliable and high-confidence genes that reveal biological phenomena underlying the etiology of disease or gene signatures prognostic of disease outcomes. This dissertation examines the capacity of graph-theoretical methods to model and analyze genomic information and thus facilitate using prior knowledge to create a more discrete and functionally relevant feature space.
    [Show full text]
  • The Peiminine Stimulating Autophagy in Human Colorectal Carcinoma Cells Via AMPK Pathway by SQSTM1
    Open Life Sci. 2016; 11: 358–366 Topical Issue on Cancer Signaling, Metastasis and Target Therapy Open Access Research Article Zhi Zheng†, Qinsi He†, Liting Xu†, Wenhao Cui, Hua Bai, Zhe Zhang, Jun Rao, Fangfang Dou* The peiminine stimulating autophagy in human colorectal carcinoma cells via AMPK pathway by SQSTM1 DOI 10.1515/biol-2016-0047 Keywords: peiminine, autophagy, natural product, Received August 19, 2016; accepted October 3, 2016 autophagic cell death, SQTEM1, AMPK/mTOC/ULK Abstract: Autophagy is a conserved catabolic process, signaling pathway. which functions in maintenance of cellular homeostasis in eukaryotic cells. The self-eating process engulfs cellular long-lived proteins and organelles with 1 Introduction double-membrane vesicles, and forms a so-called autophagosome. Degradation of contents via fusion Autophagy is an evolutionarily conserved cellular with lysosome provides recycled building blocks for pathway that delivers cellular contents to lysosomes synthesis of new molecules during stress, e.g. starvation. for degradation. Three types of autophagy have Peiminine is a steroidal alkaloid extracted from Fritillaria been identified as chaperone-mediated autophagy, thunbergii which is widely used in Traditional Chinese microautophagy and macroautophagy [1]. Autophagy is Medicine. Previously, peiminine has been identified to an important regulatory process in eukaryotic cells for induce autophagy in human colorectal carcinoma cells. removing long-lived molecules and organelles; during In this study, we further investigated whether peiminine autophagy, autophagosomes are formed, engulfed with could induce autophagic cell death via activating cytoplasm and organelles, and followed by fusion with autophagy-related signaling pathway AMPK-mTOR-ULK lysosomes for degradation. The degraded products were by promoting SQSTM1(P62).
    [Show full text]
  • Research Article Apoptotic Cell Death in the Lactating Mammary Gland Is
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Bern Open Repository and Information System (BORIS) CMLS, Cell. Mol. Life Sci. 61 (2004) 1221–1228 1420-682X/04/101221-08 DOI 10.1007/s00018-004-4046-7 CMLS Cellular and Molecular Life Sciences © Birkhäuser Verlag, Basel, 2004 Research Article Apoptotic cell death in the lactating mammary gland is enhanced by a folding variant of a-lactalbumin A. Baltzer a,C.Svanborg b and R. Jaggi a,* a Department of Clinical Research, University of Bern, Murtenstrasse 35, 3010 Bern (Switzerland), Fax: +41 31 632 32 97, e-mail: [email protected] b Lund University, Division of Microbiology, Immunology and Glycobiology, Lund, 223 62 (Sweden) Received 30 January 2004; received after revision 5 March 2004; accepted 16 March 2004 Abstract. Apoptosis is essential to eliminate secretory pase-3 activity in alveolar epithelial cells near the HAM- epithelial cells during the involution of the mammary LET pellets but not more distant to the pellet or in con- gland. The environmental regulation of this process is tralateral glands. The effect was specific for HAMLET however, poorly understood. This study tested the effect and no effects were observed when mammary glands of HAMLET (human a-lactalbumin made lethal to tumor were exposed to native a-lactalbumin or fatty acid alone. cells) on mammary cells. Plastic pellets containing HAMLET also induced cell death in vitro in a mouse HAMLET were implanted into the fourth inguinal mam- mammary epithelial cell line. The results suggest that mary gland of lactating mice for 3 days.
    [Show full text]