DOCSLIB.ORG

Original Article PFTK1 Promotes the Progression of Non-Small Cell Lung Cancer (NSCLC) Through the Wnt/Β-Catenin Signaling Pathway

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Original Article PFTK1 Promotes the Progression of Non-Small Cell Lung Cancer (NSCLC) Through the Wnt/Β-Catenin Signaling Pathway Int J Clin Exp Pathol 2016;9(12):12228-12239 www.ijcep.com /ISSN:1936-2625/IJCEP0037998 Original Article PFTK1 promotes the progression of non-small cell lung cancer (NSCLC) through the Wnt/β-catenin signaling pathway Liang-Liang Jia1,2*, Jie Chen3*, Ling Gai4, Yu-Peng Li5, Xiao-Ning Lu6, Xue-Fan Cui1, Li-Li Ji2,7 1Department of Respiratory Medicine, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, China; 2Department of Pathology, Medical College, Nantong University, Nantong, Jiangsu, China; 3Department of Oncology, Jiangyin People’s Hospital, Jiangyin, Jiangsu, China; 4Department of Oncology, Affiliated Hospital of Nantong University, Nantong, Jiangsu, China; 5Department of Pediatrics, The People’s Hospital of Rizhao City, Rizhao, Shandong, China; 6Department of Thoracic Surgery, Affiliated Hospital of Nantong University, Nantong, Jiangsu, China; 7Department of Jiangsu Province Key Laboratory for Inflammation and Molecular Drug Target, Medical College, Nantong University, Nantong, Jiangsu, China. *Equal contributors. Received July 17, 2016; Accepted October 10, 2016; Epub December 1, 2016; Published December 15, 2016 Abstract: PFTK1, as a Cdc-related serine/threonine protein kinase, is a crucial regulator of cyclins and cell cycle. However, the role of PFTK1 in non-small cell lung cancer (NSCLC) remains elusive. In the present study, Western blot and immunohistochemistry (IHC) assays demonstrated that PFTK1 was up-regulated in lung cancer tissues and cell lines. In addition, IHC analysis revealed that PFTK1 expression was associated with histological differentiation, lymph node status, TNM stage and predicted poor prognosis of NSCLC. In vitro studies using serum starvation- refeeding experiment and PFTK1 overexpression assay demonstrated that PFTK1 expression promoted prolifera- tion of NSCLC cells, while PFTK1 knockdown assay showed that PFTK1 knockdown led to decreased cell growth rate. Besides, the transcription activity of Wnt/β-catenin pathway was enhanced when PFTK1 was overexpressed. Our finding supported that up-regulated PFTK1 might promote the progression of NSCLC via activating the Wnt/β- catenin signaling pathway. Keywords: PFTK1, NSCLC, proliferation, Wnt Introduction PFTK1 is a member of Cdc2-related serine/ threonine protein kinases, also named as Lung cancer is the leading cause of cancer- PFTAIRE1, which share a highly conserved motif related death in developed countries [7] and is PSTAIRE and are crucial regulators of cyclins becoming a common malignancy in developing and cell cycle [33]. The gene of PFTAIRE1 is countries because of air pollution and a high located at human chromosome 7q21.13, which cigarette smoking ratio, such as in China. Non- is highly expressed in the brain, pancreas, kid- small cell lung cancer (NSCLC) is the most com- ney, heart, testis, and ovary and minimally mon type of lung cancer, accounting for approx- expressed in the spleen and thymus. The func- imately 80% of all lung cancer occurrences [2]. tion of PFTK1 was recently reported as a cyclin- Despite advances in early detection and stan- dependent kinase (CDK), regulating cell cycle dard treatment, the 5-year survival rate of progression and cell proliferation by specifically patients suffering from NSCLC is merely 15%, interacting with members of cyclin proteins which largely due to the fact that most of them such as cyclin D3 (CCND3) and cyclin Y (CCNY) are diagnosed at advanced stage [3]. Complete [14]. CCNY played an important role in Droso- understanding of the molecular mechanism of phila embryogenesis [15] and was associated NSCLC will contribute to the development of with human NSCLC proliferation and tumorigen- diagnostic technologies and new treatment esis [34]. The interaction between PFTK1 and methods. CCNY was reported to corporately mediate PFTK1 promotes progression of NSCLC through Wnt/β-catenin pathway Table 1. Relationship between PFTK1 expression The Wnt pathway functions in multiple cellular and clinicopathological factors in 140 lung adeno- biological processes and is involved in various carcinoma patients human diseases including cancer [4]. The Wnt PFTK1 signaling is divided into two branches: the Parameters Total expression P canonical β-catenin-dependent pathway and the non-canonical planar cell polarity (PCP) Low High and Wnt/Ca2+ pathway [16]. The Wnt/β-catenin Age (year) 0.197 pathway is activated when a Wnt ligand binds < 60 61 32 29 to Frizzled (Fz) receptor and its co-receptor ≥ 60 79 50 29 LRP6/5. The formation of Wnt-Fz-LRP6 com- Gender 0.13 plex, together with the recruitment of the scaf- Male 69 36 33 folding protein dishevelled (Dvl), results in LRP6 Female 71 46 25 phosphorylation and activation and the recruit- Tumor size (cm) 0.106 ment of the Axin complex to the receptors [18]. < 3 62 41 21 Accumulated β-catenin in the cytoplasm travels to the nucleus and forms complexes with T cell ≥ 3 78 41 37 factor/lymphoid enhancer factor (TCF/LEF), Smoking status 0.069 and activates the Wnt target genes including Yes 20 8 12 cyclin D1 and c-myc, which results in tumori- No 120 74 46 genesis [10, 28]. Histological differentiation 0.025* Poor 17 6 11 In this study, we found that the expression of Mod 94 54 40 PFTK1 was significantly increased in NSCLC cells and surgical tissues by assays of western Well 29 22 7 blotting. We also investigated the association Lymph node status 0.001* of PFTK1 with clinical and pathological par- 0 86 63 23 ameters. In addition, PFTK1 expression was > 0 54 19 35 changed by small interference RNA to explore TNM stage 0.004* its role in the progression of NSCLC cells. I 92 64 28 Meanwhile, we also detected the role of PFTK1 II 30 12 18 in the activation of Wnt/β-catenin signaling III 15 5 10 pathway and its related protein expression, IV 3 1 2 such as β-catenin, c-myc and cyclin D1. There- fore, we hypothesized that PFTK1 could pro- Ki67 expression 0.001* mote the progression of NSCLC cell and would Low 81 65 16 be a potential treatment target of NSCLC. High 59 17 42 Note: Statistical analyses were performed by the Pearson χ2 Materials and methods test. *P < 0.05 was considered significant. Lung cancer tissue specimens and cell lines phosphorylation of low density lipoprotein Eight pairs of lung cancers and adjacent normal receptor-related protein 6 (LRP6) in Drosophila fresh tissues stored at -80°C until were ana- [6]. It was well known that phosphorylation of lyzed by Western blot. Then 140 lung adenocar- transmembrane receptor LRP6 represented an cinoma tissue specimens were obtained from initial step of the canonical Wnt signaling cas- the Department of Pathology, Affiliated Hospital cade [4, 31, 35]. It was reported that PFTK1- of Nantong University from 2003 to 2008, and CCNY complex activated non-canonical Wnt clinicopathologic data of these specimens was signaling in hepatocellular carcinoma [26] and shown in Table 1. Every patient was asked to PFTK1 played an oncogenic role in promoting participate in the study, and the removed tis- the cellular invasion and motility of HCC cells sues for research purposes were obtained from [20]. In addition, PFTK1 promoted tumor cell the Ethics Committee of Nantong University proliferation, migration and invasion in breast Cancer Hospital. No patient was treated with cancer [9]. However, whether PFTK1 affecting chemotherapy or radiotherapy. The lung cancer the progression of NSCLC remained unclear. cell lines A549, H1299 and SPCA1 were 12229 Int J Clin Exp Pathol 2016;9(12):12228-12239 PFTK1 promotes progression of NSCLC through Wnt/β-catenin pathway obtained from the Institute of Cell Biology, were as follows: anti-PFTK1 (1:100, Santa Cruz Academic of China (Shanghai, China). Biotechnology) and anti-Ki-67 (1:100, Santa Cruz Biotechnology). For assessment of PFTK1 Western blot analysis and antibodies and Ki-67, five high-power fields in each speci- men were selected randomly, and then cyto- Western blot assay was performed as previ- plasm and nuclear staining were examined. The ously described [17]. Tissue and cell protein degree of immunostaining was viewed and were homogenized in a homogenization buffer scored separately by 2 independent investiga- containing 50 mM Tris-HCL, pH 7.5, 150 mM tors without knowledge of clinicopathologic NaCl, 1% NP-40, 5 mM EDTA, 60 mM data in a blind manner. Tumor cell proportion β-glycerophosphate, 0.1 mM sodium orthovan- was scored as follows: 0 (no positive tumor adate, o.1 mM NaF, and complete protease cells); 1 (< 10% positive tumor cells); 2 (10-35% inhibitor cocktail (Roche Diagnostic), and then positive tumor cells); 3 (35-70% positive tu- placed on the ice for 0.5 h after being fully mor cells) and 4 (> 70% positive tumor cells). homogenized. Then the sample was centri- Staining intensity was graded according to the fuged at 13,000 r for 15 min to collect the following criteria: 0 (no staining); 1 (weak stain- supernatant. Protein concentrations were mea- ing = light yellow); 2 (moderate staining = yellow sured with a Bio-Rad protein assay (Bio-Rad, brown) and 3 (strong staining = brown). Staining Hercules, CA). The supernatant diluted in 2 × index (SI) was calculated as the product of SDS loading buffer and boiled for 15 min. staining intensity score and the proportion of Proteins were separated with SDS-PAGE (sodi- positive tumor cells.SI score ≥ 6 was used to um dodecyl sulfate-polyacrylamide gel electro- define tumors with the high expression of phoresis) and then transferred to polyvinyli- PFTK1, and an SI score ≤ 4 was used to indi- denedifluoride filter (PVDF) membranes (Milli- cate the low expression of PFTK1. pore, Bedford, MA). The membranes were blocked with phosphate-buffered saline (PBS) The SPSS18.0 statistical program was used for containing 0.1% Tween 20 and 5% non-fat milk statistical analysis. The Chi-square test was for 2 h and then were washed with phosphate- performed to evaluate the relationship bet- buffered saline (PBS) containing 0.1% Tween ween PFTK1 expression and clinicopathologi- 20 (PBST) for three times.
Load more

Recommended publications
  • Profiling Data
    Compound Name DiscoveRx Gene Symbol Entrez Gene Percent Compound Symbol Control Concentration (nM) JNK-IN-8 AAK1 AAK1 69 1000 JNK-IN-8 ABL1(E255K)-phosphorylated ABL1 100 1000 JNK-IN-8 ABL1(F317I)-nonphosphorylated ABL1 87 1000 JNK-IN-8 ABL1(F317I)-phosphorylated ABL1 100 1000 JNK-IN-8 ABL1(F317L)-nonphosphorylated ABL1 65 1000 JNK-IN-8 ABL1(F317L)-phosphorylated ABL1 61 1000 JNK-IN-8 ABL1(H396P)-nonphosphorylated ABL1 42 1000 JNK-IN-8 ABL1(H396P)-phosphorylated ABL1 60 1000 JNK-IN-8 ABL1(M351T)-phosphorylated ABL1 81 1000 JNK-IN-8 ABL1(Q252H)-nonphosphorylated ABL1 100 1000 JNK-IN-8 ABL1(Q252H)-phosphorylated ABL1 56 1000 JNK-IN-8 ABL1(T315I)-nonphosphorylated ABL1 100 1000 JNK-IN-8 ABL1(T315I)-phosphorylated ABL1 92 1000 JNK-IN-8 ABL1(Y253F)-phosphorylated ABL1 71 1000 JNK-IN-8 ABL1-nonphosphorylated ABL1 97 1000 JNK-IN-8 ABL1-phosphorylated ABL1 100 1000 JNK-IN-8 ABL2 ABL2 97 1000 JNK-IN-8 ACVR1 ACVR1 100 1000 JNK-IN-8 ACVR1B ACVR1B 88 1000 JNK-IN-8 ACVR2A ACVR2A 100 1000 JNK-IN-8 ACVR2B ACVR2B 100 1000 JNK-IN-8 ACVRL1 ACVRL1 96 1000 JNK-IN-8 ADCK3 CABC1 100 1000 JNK-IN-8 ADCK4 ADCK4 93 1000 JNK-IN-8 AKT1 AKT1 100 1000 JNK-IN-8 AKT2 AKT2 100 1000 JNK-IN-8 AKT3 AKT3 100 1000 JNK-IN-8 ALK ALK 85 1000 JNK-IN-8 AMPK-alpha1 PRKAA1 100 1000 JNK-IN-8 AMPK-alpha2 PRKAA2 84 1000 JNK-IN-8 ANKK1 ANKK1 75 1000 JNK-IN-8 ARK5 NUAK1 100 1000 JNK-IN-8 ASK1 MAP3K5 100 1000 JNK-IN-8 ASK2 MAP3K6 93 1000 JNK-IN-8 AURKA AURKA 100 1000 JNK-IN-8 AURKA AURKA 84 1000 JNK-IN-8 AURKB AURKB 83 1000 JNK-IN-8 AURKB AURKB 96 1000 JNK-IN-8 AURKC AURKC 95 1000 JNK-IN-8
    [Show full text]
  • Anti-CDK14 / PFTK1 Antibody (ARG40789)
    Product datasheet [email protected] ARG40789 Package: 100 μl anti-CDK14 / PFTK1 antibody Store at: -20°C Summary Product Description Rabbit Polyclonal antibody recognizes CDK14 / PFTK1 Tested Reactivity Hu Tested Application FACS, IHC-P, WB Host Rabbit Clonality Polyclonal Isotype IgG Target Name CDK14 / PFTK1 Antigen Species Human Immunogen KLH-conjugated synthetic peptide corresponding to aa. 67-96 of Human CDK14 / PFTK1. Conjugation Un-conjugated Alternate Names PFTAIRE1; Cell division protein kinase 14; hPFTAIRE1; EC 2.7.11.22; Cyclin-dependent kinase 14; PFTK1; Serine/threonine-protein kinase PFTAIRE-1 Application Instructions Application table Application Dilution FACS 1:10 - 1:50 IHC-P 1:50 - 1:100 WB 1:1000 Application Note * The dilutions indicate recommended starting dilutions and the optimal dilutions or concentrations should be determined by the scientist. Calculated Mw 53 kDa Properties Form Liquid Purification Saturated Ammonium Sulfate (SAS) precipitation followed by dialysis against PBS. Buffer PBS and 0.09% (W/V) Sodium azide. Preservative 0.09% (W/V) Sodium azide. Storage instruction For continuous use, store undiluted antibody at 2-8°C for up to a week. For long-term storage, aliquot and store at -20°C or below. Storage in frost free freezers is not recommended. Avoid repeated freeze/thaw cycles. Suggest spin the vial prior to opening. The antibody solution should be gently mixed before use. Note For laboratory research only, not for drug, diagnostic or other use. www.arigobio.com 1/3 Bioinformation Gene Symbol CDK14 Gene Full Name cyclin-dependent kinase 14 Background PFTK1 is a member of the CDC2 (MIM 116940)-related protein kinase family (Yang and Chen, 2001 [PubMed 11313143]).[supplied by OMIM, Mar 2008] Function Serine/threonine-protein kinase involved in the control of the eukaryotic cell cycle, whose activity is controlled by an associated cyclin.
    [Show full text]
  • Delineation of Key Regulatory Elements Identifies Points Of
    DELINEATION OF KEY REGULATORY ELEMENTS IDENTIFIES POINTS OF VULNERABILITY IN THE MITOGEN-ACTIVATED SIGNALING NETWORK SUPPLEMENTARY MATERIALS List of contents Supplementary Figures with legends 1. Figure S1: Distribution of primary siRNA screen data, and standardization of assay procedure. 2. Figure S2: Scatter plot of screen data. 3. Figure S3: Functional relevance of the identified targets and Calculation of residence time from PDT and cell cycle distribution. 4. Figure S4: FACS profiles for ABL1 and AKT1. Table for data in Figure 5B. 5. Figure S5: Venn diagram showing the results of the comparative analysis of other screen results 6. Figure S6: Dose response profiles for the AKT1 + ABL1 inhibitor combination for CH1, list of the 14 cell lines and their description, effect of ABL1+AKT1 inhibitor combination on increase in apoptotic cells and G1 arrest in 14 cell lines, effects of CHEK1 inhibitor on combination C1,C2 on 4 cell lines. Supplementary Tables 1. Table S1: siRNA screen results for targeted kinases and phosphatases. 2. Table S2: Gene expression status of the validated hits. 3. Table S3: Role played by identified RNAi hits in regulation of cell cycle, the effect on PDTs along with phase-specific RTs. 4. Table S4: List of molecules classified as cell cycle targets. 5. Table S5: High confidence network used for graph theory analysis. 6. Table S6: Occurrences of nodes in shortest path networks. 7. Table S7: Network file used as SNAVI background. 8. Table S8: Classification of nodes present in modules according to specificity. Legends for tables Supplementary Experimental Procedures References Figure S1 A 450 400 G1 S 350 G2 300 250 200 150 100 50 Distribution of molecules Distribution 0 -6-4-20246 Z-score 350 200 400 G1 S 300 G2 150 300 250 200 100 200 150 100 50 100 Distribution of molecules 50 0 0 0 -4 -2 0 2 4 -4-20246 -4-20246 Z-score B PLK1 GAPDH PLCg BTK PLCg CDC2A PLCg CHEK1 PLCg MET Distribution profiles of complete primary screen and western blots showing knockdown efficiency.
    [Show full text]
  • Downregulation of Carnitine Acyl-Carnitine Translocase by Mirnas
    Page 1 of 288 Diabetes 1 Downregulation of Carnitine acyl-carnitine translocase by miRNAs 132 and 212 amplifies glucose-stimulated insulin secretion Mufaddal S. Soni1, Mary E. Rabaglia1, Sushant Bhatnagar1, Jin Shang2, Olga Ilkayeva3, Randall Mynatt4, Yun-Ping Zhou2, Eric E. Schadt6, Nancy A.Thornberry2, Deborah M. Muoio5, Mark P. Keller1 and Alan D. Attie1 From the 1Department of Biochemistry, University of Wisconsin, Madison, Wisconsin; 2Department of Metabolic Disorders-Diabetes, Merck Research Laboratories, Rahway, New Jersey; 3Sarah W. Stedman Nutrition and Metabolism Center, Duke Institute of Molecular Physiology, 5Departments of Medicine and Pharmacology and Cancer Biology, Durham, North Carolina. 4Pennington Biomedical Research Center, Louisiana State University system, Baton Rouge, Louisiana; 6Institute for Genomics and Multiscale Biology, Mount Sinai School of Medicine, New York, New York. Corresponding author Alan D. Attie, 543A Biochemistry Addition, 433 Babcock Drive, Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin, (608) 262-1372 (Ph), (608) 263-9608 (fax), [email protected]. Running Title: Fatty acyl-carnitines enhance insulin secretion Abstract word count: 163 Main text Word count: 3960 Number of tables: 0 Number of figures: 5 Diabetes Publish Ahead of Print, published online June 26, 2014 Diabetes Page 2 of 288 2 ABSTRACT We previously demonstrated that micro-RNAs 132 and 212 are differentially upregulated in response to obesity in two mouse strains that differ in their susceptibility to obesity-induced diabetes. Here we show the overexpression of micro-RNAs 132 and 212 enhances insulin secretion (IS) in response to glucose and other secretagogues including non-fuel stimuli. We determined that carnitine acyl-carnitine translocase (CACT, Slc25a20) is a direct target of these miRNAs.
    [Show full text]
  • Targeting Cyclin-Dependent Kinases in Human Cancers: from Small Molecules to Peptide Inhibitors
    Cancers 2015, 7, 179-237; doi:10.3390/cancers7010179 OPEN ACCESS cancers ISSN 2072-6694 www.mdpi.com/journal/cancers Review Targeting Cyclin-Dependent Kinases in Human Cancers: From Small Molecules to Peptide Inhibitors Marion Peyressatre †, Camille Prével †, Morgan Pellerano and May C. Morris * Institut des Biomolécules Max Mousseron, IBMM-CNRS-UMR5247, 15 Av. Charles Flahault, 34093 Montpellier, France; E-Mails: [email protected] (M.P.); [email protected] (C.P.); [email protected] (M.P.) † These authors contributed equally to this work. * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +33-04-1175-9624; Fax: +33-04-1175-9641. Academic Editor: Jonas Cicenas Received: 17 December 2014 / Accepted: 12 January 2015 / Published: 23 January 2015 Abstract: Cyclin-dependent kinases (CDK/Cyclins) form a family of heterodimeric kinases that play central roles in regulation of cell cycle progression, transcription and other major biological processes including neuronal differentiation and metabolism. Constitutive or deregulated hyperactivity of these kinases due to amplification, overexpression or mutation of cyclins or CDK, contributes to proliferation of cancer cells, and aberrant activity of these kinases has been reported in a wide variety of human cancers. These kinases therefore constitute biomarkers of proliferation and attractive pharmacological targets for development of anticancer therapeutics. The structural features of several of these kinases have been elucidated and their molecular mechanisms of regulation characterized in depth, providing clues for development of drugs and inhibitors to disrupt their function. However, like most other kinases, they constitute a challenging class of therapeutic targets due to their highly conserved structural features and ATP-binding pocket.
    [Show full text]
  • Oxidized Phospholipids Regulate Amino Acid Metabolism Through MTHFD2 to Facilitate Nucleotide Release in Endothelial Cells
    ARTICLE DOI: 10.1038/s41467-018-04602-0 OPEN Oxidized phospholipids regulate amino acid metabolism through MTHFD2 to facilitate nucleotide release in endothelial cells Juliane Hitzel1,2, Eunjee Lee3,4, Yi Zhang 3,5,Sofia Iris Bibli2,6, Xiaogang Li7, Sven Zukunft 2,6, Beatrice Pflüger1,2, Jiong Hu2,6, Christoph Schürmann1,2, Andrea Estefania Vasconez1,2, James A. Oo1,2, Adelheid Kratzer8,9, Sandeep Kumar 10, Flávia Rezende1,2, Ivana Josipovic1,2, Dominique Thomas11, Hector Giral8,9, Yannick Schreiber12, Gerd Geisslinger11,12, Christian Fork1,2, Xia Yang13, Fragiska Sigala14, Casey E. Romanoski15, Jens Kroll7, Hanjoong Jo 10, Ulf Landmesser8,9,16, Aldons J. Lusis17, 1234567890():,; Dmitry Namgaladze18, Ingrid Fleming2,6, Matthias S. Leisegang1,2, Jun Zhu 3,4 & Ralf P. Brandes1,2 Oxidized phospholipids (oxPAPC) induce endothelial dysfunction and atherosclerosis. Here we show that oxPAPC induce a gene network regulating serine-glycine metabolism with the mitochondrial methylenetetrahydrofolate dehydrogenase/cyclohydrolase (MTHFD2) as a cau- sal regulator using integrative network modeling and Bayesian network analysis in human aortic endothelial cells. The cluster is activated in human plaque material and by atherogenic lipo- proteins isolated from plasma of patients with coronary artery disease (CAD). Single nucleotide polymorphisms (SNPs) within the MTHFD2-controlled cluster associate with CAD. The MTHFD2-controlled cluster redirects metabolism to glycine synthesis to replenish purine nucleotides. Since endothelial cells secrete purines in response to oxPAPC, the MTHFD2- controlled response maintains endothelial ATP. Accordingly, MTHFD2-dependent glycine synthesis is a prerequisite for angiogenesis. Thus, we propose that endothelial cells undergo MTHFD2-mediated reprogramming toward serine-glycine and mitochondrial one-carbon metabolism to compensate for the loss of ATP in response to oxPAPC during atherosclerosis.
    [Show full text]
  • Gene Symbol Accession Alias/Prev Symbol Official Full Name AAK1 NM 014911.2 KIAA1048, Dkfzp686k16132 AP2 Associated Kinase 1
    Gene Symbol Accession Alias/Prev Symbol Official Full Name AAK1 NM_014911.2 KIAA1048, DKFZp686K16132 AP2 associated kinase 1 (AAK1) AATK NM_001080395.2 AATYK, AATYK1, KIAA0641, LMR1, LMTK1, p35BP apoptosis-associated tyrosine kinase (AATK) ABL1 NM_007313.2 ABL, JTK7, c-ABL, p150 v-abl Abelson murine leukemia viral oncogene homolog 1 (ABL1) ABL2 NM_007314.3 ABLL, ARG v-abl Abelson murine leukemia viral oncogene homolog 2 (arg, Abelson-related gene) (ABL2) ACVR1 NM_001105.2 ACVRLK2, SKR1, ALK2, ACVR1A activin A receptor ACVR1B NM_004302.3 ACVRLK4, ALK4, SKR2, ActRIB activin A receptor, type IB (ACVR1B) ACVR1C NM_145259.2 ACVRLK7, ALK7 activin A receptor, type IC (ACVR1C) ACVR2A NM_001616.3 ACVR2, ACTRII activin A receptor ACVR2B NM_001106.2 ActR-IIB activin A receptor ACVRL1 NM_000020.1 ACVRLK1, ORW2, HHT2, ALK1, HHT activin A receptor type II-like 1 (ACVRL1) ADCK1 NM_020421.2 FLJ39600 aarF domain containing kinase 1 (ADCK1) ADCK2 NM_052853.3 MGC20727 aarF domain containing kinase 2 (ADCK2) ADCK3 NM_020247.3 CABC1, COQ8, SCAR9 chaperone, ABC1 activity of bc1 complex like (S. pombe) (CABC1) ADCK4 NM_024876.3 aarF domain containing kinase 4 (ADCK4) ADCK5 NM_174922.3 FLJ35454 aarF domain containing kinase 5 (ADCK5) ADRBK1 NM_001619.2 GRK2, BARK1 adrenergic, beta, receptor kinase 1 (ADRBK1) ADRBK2 NM_005160.2 GRK3, BARK2 adrenergic, beta, receptor kinase 2 (ADRBK2) AKT1 NM_001014431.1 RAC, PKB, PRKBA, AKT v-akt murine thymoma viral oncogene homolog 1 (AKT1) AKT2 NM_001626.2 v-akt murine thymoma viral oncogene homolog 2 (AKT2) AKT3 NM_181690.1
    [Show full text]
  • Technische Universität München
    TECHNISCHE UNIVERSITÄT MÜNCHEN Lehrstuhl für Proteomik und Bioanalytik Application of mass spectrometry-based proteomics to study cancer drug resistance mechanisms Heiner Matthias Koch Vollständiger Abdruck der von der Fakultät Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt der Technischen Universität München zur Erlangung des akademischen Grades eines Doktors der Naturwissenschaften genehmigten Dissertation. Vorsitzender: Prof. Dr. D. Langosch Prüfer der Dissertation: 1. Prof. Dr. B. Küster 2. Prof. Dr. F. Bassermann Die Dissertation wurde am 12.07.2016 bei der Technischen Universität München eingereicht und durch die Fakultät Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt am 20.09.2016 angenommen. II Table of Content Abstract V Zusammenfassung VI Chapter I General Introduction 1 Chapter II Chemical proteomics uncovers EPHA2 as a mechanism 49 of acquired resistance to small molecule EGFR kinase inhibition Chapter III Phosphoproteome profiling reveals molecular mechanisms of 73 growth factor mediated kinase inhibitor resistance in EGFR overexpressing cancer cells Chapter IV Time resolved proteomic and phosphoproteomic analysis of 109 adaptation to kinase inhibition Chapter V General discussion 135 List of publications 144 Danksagung | Acknowledgment 145 Curriculum vitae 146 III IV Abstract In recent years an increasing number of small molecule kinase inhibitors were approved for targeted cancer therapies. Targeted therapies have less toxic side effects than conventional chemotherapeutics and promise efficacious personalized treatments. Although some molecules improved the outcome of selected patient groups, resistance almost invariably develops and represents a major clinical challenge. There is an intensive effort to circumvent emerging resistance by the development of new targeted agents or the combination of approved molecules. However, the molecular alterations that render cancer cells resistant are still poorly understood.
    [Show full text]
  • A Novel Interplay Between Oncogenic PFTK1 Protein Kinase and Tumor Suppressor TAGLN2 in the Control of Liver Cancer Cell Motility
    Oncogene (2011) 30, 4464–4475 & 2011 Macmillan Publishers Limited All rights reserved 0950-9232/11 www.nature.com/onc ORIGINAL ARTICLE A novel interplay between oncogenic PFTK1 protein kinase and tumor suppressor TAGLN2 in the control of liver cancer cell motility WKC Leung1,2, AKK Ching1,2, AWH Chan1, TCW Poon2,3, H Mian1,2, AST Wong4, K-F To1,2 and N Wong1,2 1Department of Anatomical and Cellular Pathology at the Li Ka-Shing Institute of Health Sciences, Chinese University of Hong Kong, Shatin, Hong Kong, China; 2State Key Laboratory in Oncology in South China, Chinese University of Hong Kong, Shatin, Hong Kong, China; 3Department of Medicine and Therapeutics, Chinese University of Hong Kong, Shatin, Hong Kong, China and 4School of Biological Sciences, University of Hong Kong, Pokfulam, Hong Kong, China The PFTK1 gene encodes a cdc2-related serine/threonine oncogene-tumor suppressor interplay, where oncogenic protein kinase that has been shown to confer cell PFTK1 confers HCC cell motility through inactivating migratory properties in hepatocellular carcinoma the actin-binding motile suppressing function of TAGLN2 (HCC). However, the prognostic value and biological via phosphorylation. mechanism by which PFTK1 promotes HCC motility Oncogene (2011) 30, 4464–4475; doi:10.1038/onc.2011.161; remain largely unknown. Here, we showed from tissue published online 16 May 2011 microarray that common upregulations of PFTK1 in primary HCC tumors (n ¼ 133/180) correlated signifi- Keywords: PFTK1; hepatocellular carcinoma; cell cantly with early age onset (p40 years), advance tumor migration; TAGLN2; actin grading and presence of microvascular invasion (Pp0.05). To understand downstream phosphorylated substrate(s) of PFTK1, phospho-proteins in PFTK1 expressing and knockdown Hep3B cells were profiled by two-dimen- Introduction sional-polyacrylamide gel electrophoresis mass spectro- metric analysis.
    [Show full text]
  • Kinome Expression Profiling to Target New Therapeutic Avenues in Multiple Myeloma
    Plasma Cell DIsorders SUPPLEMENTARY APPENDIX Kinome expression profiling to target new therapeutic avenues in multiple myeloma Hugues de Boussac, 1 Angélique Bruyer, 1 Michel Jourdan, 1 Anke Maes, 2 Nicolas Robert, 3 Claire Gourzones, 1 Laure Vincent, 4 Anja Seckinger, 5,6 Guillaume Cartron, 4,7,8 Dirk Hose, 5,6 Elke De Bruyne, 2 Alboukadel Kassambara, 1 Philippe Pasero 1 and Jérôme Moreaux 1,3,8 1IGH, CNRS, Université de Montpellier, Montpellier, France; 2Department of Hematology and Immunology, Myeloma Center Brussels, Vrije Universiteit Brussel, Brussels, Belgium; 3CHU Montpellier, Laboratory for Monitoring Innovative Therapies, Department of Biologi - cal Hematology, Montpellier, France; 4CHU Montpellier, Department of Clinical Hematology, Montpellier, France; 5Medizinische Klinik und Poliklinik V, Universitätsklinikum Heidelberg, Heidelberg, Germany; 6Nationales Centrum für Tumorerkrankungen, Heidelberg , Ger - many; 7Université de Montpellier, UMR CNRS 5235, Montpellier, France and 8 Université de Montpellier, UFR de Médecine, Montpel - lier, France ©2020 Ferrata Storti Foundation. This is an open-access paper. doi:10.3324/haematol. 2018.208306 Received: October 5, 2018. Accepted: July 5, 2019. Pre-published: July 9, 2019. Correspondence: JEROME MOREAUX - [email protected] Supplementary experiment procedures Kinome Index A list of 661 genes of kinases or kinases related have been extracted from literature9, and challenged in the HM cohort for OS prognostic values The prognostic value of each of the genes was computed using maximally selected rank test from R package MaxStat. After Benjamini Hochberg multiple testing correction a list of 104 significant prognostic genes has been extracted. This second list has then been challenged for similar prognosis value in the UAMS-TT2 validation cohort.
    [Show full text]
  • Dissertation Submitted to the Combined Faculties for the Natural
    Dissertation submitted to the Combined Faculties for the Natural Sciences and for Mathematics of the Ruperto-Carola University of Heidelberg, Germany for the degree of Doctor of Natural Sciences presented by Diplom-Biochemiker Johannes Hermle born in: Offenbach a.M., Germany Oral-examination: July 26, 2017 siRNA SCREEN FOR IDENTIFICATION OF HUMAN KINASES INVOLVED IN ASSEMBLY AND RELEASE OF HIV-1 Referees: Prof. Dr. Hans-Georg Kräusslich Prof. Dr. Dirk Grimm ii Meiner Familie iii Summary Summary The replication of the human immunodeficiency virus type 1 (HIV-1) is as yet not fully understood. In particular the knowledge of interactions between viral and host cell proteins and the understanding of complete virus-host protein networks are still imprecise. An integral picture of the hijacked cellular machinery is essential for a better comprehension of the virus. And as a prerequisite, new tools are needed for this purpose. To create such a novel tool, a screening platform for host cell factors was established in this work. The screening assay serves as a powerful method to gain insights into virus-host-interactions. It was specifically tailored to addressing the stage of assembly and release of viral particles during the replication cycle of HIV-1. It was designed to be suitable for both RNAi and chemical compound screening. The first phase of this work comprised the setup and optimization of the assay. It was shown, that it was robust and reliable and delivered reproducible results. As a subsequent step, a siRNA library targeting 724 human kinases and accessory proteins was examined. After the evaluation of the complete siRNA library in a primary screen, all primary hits were validated in a second reconfirmation screen using different siRNAs.
    [Show full text]
  • Supplementary Data
    Human Breast Tumor Acquisition S1 Human Breast Tumor Samples Analysis of 102 human breast tumor samples 58 ER-positive, 44 ER-negative, 24 “triple negative” Affymetrix Microarray data analyzed for kinase expression 86 kinases or kinase-associated genes differentially expressed between ER-negative and ER-positive samples. 52 genes identified as being at least 2-fold higher in ER-negative tumors (permutation p-value <.05) Validation of kinases identified in kinase expression array analysis Validation using publicly Validation done in an Validation done in ER- available data sets of independent set of human negative and ER-positive human breast cancers breast tumors using Q-RT-PCR breast cancer cell lines profiled using gene analysis using Affymetrix expression profiling expression profiling data and Q-RT-PCR analysis Kinase knockdown of differentially expressed, validated kinases or kinase- associated genes to assess effect on growth of ER-negative and ER-positive breast cancer cells Identified kinases that are critical for the growth of ER-negative breast cancer S2 Immune cell infiltration score Cell Cycle CheckpointCell Cycle S6 kinase P -value: 0.62 -value: MAPK Immunomodulatory Immune cell infiltration score P -value: >0.5 for all groups compared against oneanother against compared allgroups >0.5 for -value: A S3 V V O A A 5-1 5-30 74 83 49 0 D 75-B MPE A-MB361 A-MB175- A-MB453 A-MB415 F7 A-MB157 A-MB436 A MB-468 F-12 F10 A MB-231 A-MB435 A-MB134- C1428 C1007 C202 C2185 4475 C-38 C1187 C1569 C1143 C2157 C70 C1008 C1599 C1937 C3153 C1500
    [Show full text]