Proteomic analysis of A2780/S ovarian cancer cell response to the cytotoxic organogold(III) compound Aubipy(c).

UNLABELLED Aubipyc is an organogold(III) compound endowed with encouraging anti-proliferative properties in vitro that is being evaluated pre-clinically as a prospective anticancer agent. A classical proteomic approach is exploited here to elucidate the mechanisms of its biological actions in A2780 human ovarian cancer cells. Based on 2-D gel electrophoresis separation and subsequent mass spectrometry identification, a considerable number of differentially expressed proteins were highlighted in A2780 cancer cells treated with Aubipyc. Bioinformatic analysis of the groups of up-regulated and down-regulated proteins pointed out that Aubipyc primarily perturbs mitochondrial processes and the glycolytic pathway. Notably, some major alterations in the glycolytic pathway were validated through Western blot and metabolic investigations. BIOLOGICAL SIGNIFICANCE This is the first proteomic analysis regarding Aubipyc cytotoxicity in A2780/S ovarian cancer cell line. Aubipyc is a promising gold(III) compound which manifests an appreciable cytotoxicity toward the cell line A2780, being able to overcome resistance to platinum. The proteomic study revealed for Aubipyc different cellular alterations with respect to cisplatin as well as to other gold compound such as auranofin. Remarkably, the bioinformatic analysis of proteomic data pointed out that Aubipyc treatment affected, directly or indirectly, several glycolytic enzymes. These data suggest a new mechanism of action for this gold drug and might have an impact on the use of gold-based drug in cancer treatment.

[1]  J. C. Bakowska,et al.  SPG20 Protein Spartin Associates with Cardiolipin via Its Plant-Related Senescence Domain and Regulates Mitochondrial Ca2+ Homeostasis , 2011, PloS one.

[2]  E. Rugarli,et al.  Loss of Prohibitin Membrane Scaffolds Impairs Mitochondrial Architecture and Leads to Tau Hyperphosphorylation and Neurodegeneration , 2012, PLoS genetics.

[3]  A. Bindoli,et al.  Induction of mitochondrial permeability transition by auranofin, a Gold(I)‐phosphine derivative , 2002, British journal of pharmacology.

[4]  Dae-Myung Jue,et al.  Gold compound auranofin inhibits IκB kinase (IKK) by modifying Cys-179 of IKKβ subunit , 2003, Experimental & Molecular Medicine.

[5]  A. Bosserhoff,et al.  Death inducer-obliterator 1 (Dido1) is a BMP target gene and promotes BMP-induced melanoma progression , 2013, Oncogene.

[6]  L. Messori,et al.  Gold(III) complexes as potential antitumor agents: solution chemistry and cytotoxic properties of some selected gold(III) compounds. , 2000, Journal of medicinal chemistry.

[7]  Ying Wang,et al.  Proteomic Approaches in Understanding Action Mechanisms of Metal-Based Anticancer Drugs , 2008, Metal-based drugs.

[8]  Sanjeev Banerjee,et al.  Retracted: Cisplatin‐induced antitumor activity is potentiated by the soy isoflavone genistein in BxPC‐3 pancreatic tumor xenografts , 2006, Cancer.

[9]  A. Modesti,et al.  Proteomic analysis of ovarian cancer cell responses to cytotoxic gold compounds. , 2012, Metallomics.

[10]  L. Messori,et al.  Gold(III) complexes with bipyridyl ligands: solution chemistry, cytotoxicity, and DNA binding properties. , 2002, Journal of medicinal chemistry.

[11]  L. Pan,et al.  Identification of platinum-resistance associated proteins through proteomic analysis of human ovarian cancer cells and their platinum-resistant sublines. , 2007, Journal of proteome research.

[12]  H. Mewes,et al.  PPI spider: A tool for the interpretation of proteomics data in the context of protein–protein interaction networks , 2009, Proteomics.

[13]  K. B. Garbutcheon-Singh,et al.  Transition metal based anticancer drugs. , 2011, Current topics in medicinal chemistry.

[14]  A. Bergamo,et al.  Molecular structure, solution chemistry and biological properties of the novel [ImH][trans-IrCl(4)(Im)(DMSO)], (I) and of the orange form of [(DMSO)(2)H][trans-IrCl(4)(DMSO)(2)], (II), complexes. , 2003, Journal of inorganic biochemistry.

[15]  D. Scudiero,et al.  New colorimetric cytotoxicity assay for anticancer-drug screening. , 1990, Journal of the National Cancer Institute.

[16]  Kathryn S Lilley,et al.  Impact of replicate types on proteomic expression analysis. , 2005, Journal of proteome research.

[17]  A. Casini,et al.  Structural characterization, solution studies, and DFT calculations on a series of binuclear gold(III) oxo complexes: relationships to biological properties. , 2008, Inorganic chemistry.

[18]  L. Messori,et al.  Interactions of selected gold(III) complexes with calf thymus DNA. , 2001, Biochemical and biophysical research communications.

[19]  John D. Storey,et al.  Statistical significance for genomewide studies , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[20]  P. Modesti,et al.  2D-DIGE analysis of ovarian cancer cell responses to cytotoxic gold compounds. , 2012, Molecular bioSystems.

[21]  Shiwei Song,et al.  A role for the mitochondrial deacetylase Sirt3 in regulating energy homeostasis , 2008, Proceedings of the National Academy of Sciences.

[22]  H. Izumi,et al.  Transcription factors and drug resistance. , 2005, European journal of cancer.

[23]  A. Casini,et al.  Gold(III) compounds as anticancer drugs , 2007 .

[24]  A. Modesti,et al.  Exploring the biochemical mechanisms of cytotoxic gold compounds: a proteomic study , 2010, JBIC Journal of Biological Inorganic Chemistry.

[25]  J. K. Kundu,et al.  Tob1 induces apoptosis and inhibits proliferation, migration and invasion of gastric cancer cells by activating Smad4 and inhibiting β-catenin signaling , 2012, International journal of oncology.

[26]  J. Marlin,et al.  Caspase Activation of p21-Activated Kinase 2 Occurs During Cisplatin-Induced Apoptosis of SH-SY5Y Neuroblastoma Cells and in SH-SY5Y Cell Culture Models of Alzheimer’s and Parkinson’s Disease , 2010 .

[27]  L. Kèlland,et al.  The resurgence of platinum-based cancer chemotherapy , 2007, Nature Reviews Cancer.

[28]  M. Gottesman,et al.  SIRT1 Contributes in Part to Cisplatin Resistance in Cancer Cells by Altering Mitochondrial Metabolism , 2008, Molecular Cancer Research.

[29]  S. Berners‐Price,et al.  Targeting the mitochondrial cell death pathway with gold compounds , 2007 .

[30]  Sahand Khakabimamaghani,et al.  CytoKavosh: A Cytoscape Plug-In for Finding Network Motifs in Large Biological Networks , 2012, PloS one.

[31]  I. Herr,et al.  Cellular stress response and apoptosis in cancer therapy. , 2001, Blood.

[32]  Renée L. Brost,et al.  The interaction network of the chaperonin CCT , 2008, The EMBO journal.

[33]  Aless,et al.  New Insights into the Molecular Mechanisms of Selected Anticancer MetalCompounds through Bioinformatic Analysis of Proteomic Data , 2013 .

[34]  Pier Giorgio Righetti,et al.  A proteomic approach to cisplatin resistance in the cervix squamous cell carcinoma cell line A431 , 2004, Proteomics.

[35]  E. Tiligada.  Chemotherapy: induction of stress responses. , 2006, Endocrine-related cancer.

[36]  L. Messori,et al.  Gold complexes inhibit mitochondrial thioredoxin reductase: consequences on mitochondrial functions. , 2004, Journal of inorganic biochemistry.

[37]  L. Ronconi,et al.  Gold dithiocarbamate derivatives as potential antineoplastic agents: design, spectroscopic properties, and in vitro antitumor activity. , 2005, Inorganic chemistry.

[38]  A. Vergara,et al.  The mode of action of anticancer gold-based drugs: a structural perspective. , 2013, Chemical communications.

[39]  D. Wessel,et al.  A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids. , 1984, Analytical biochemistry.

[40]  I. Ott.  On the medicinal chemistry of gold complexes as anticancer drugs , 2009 .

[41]  L. Massai,et al.  Chemistry and biology of three representative gold(III) compounds as prospective anticancer agents , 2012 .

[42]  M. Wilkins,et al.  Optimal replication and the importance of experimental design for gel-based quantitative proteomics. , 2005, Journal of proteome research.

[43]  S. Maehara,et al.  Proteomics finding heat shock protein 27 as a biomarker for resistance of pancreatic cancer cells to gemcitabine. , 2007, International journal of oncology.

[44]  Jennifer L. O'Day.  Statistical Significance for Genome Wide Studies Under Unequal Variance , 2015 .

[45]  H. Chung,et al.  Metabolic approaches to overcoming chemoresistance in ovarian cancer , 2011, Annals of the New York Academy of Sciences.

[46]  C. Che,et al.  The anti-cancer properties of gold(III) compounds with dianionic porphyrin and tetradentate ligands , 2009 .

[47]  P. Drew,et al.  The proteomic analysis of cisplatin resistance in breast cancer cells. , 2007, Oncology research.

[48]  Brad T. Sherman,et al.  Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists , 2008, Nucleic acids research.

[49]  A. Casini,et al.  Gold compounds as anticancer agents: chemistry, cellular pharmacology, and preclinical studies , 2010, Medicinal research reviews.

[50]  E. Yim,et al.  Analysis of differential protein expression by cisplatin treatment in cervical carcinoma cells , 2005, International Journal of Gynecologic Cancer.

[51]  L. Messori,et al.  Solution chemistry and cytotoxic properties of novel organogold(III) compounds. , 2004, Bioorganic & medicinal chemistry.

[52]  S. Choudhary,et al.  Inducible Tumor Necrosis Factor (TNF) Receptor-associated Factor-1 Expression Couples the Canonical to the Non-canonical NF-κB Pathway in TNF Stimulation* , 2013, The Journal of Biological Chemistry.

[53]  S. Oliver,et al.  Chromatographic separations as a prelude to two‐dimensional electrophoresis in proteomics analysis , 2001, Proteomics.

[54]  Andrea Urbani,et al.  A proteomic approach to paclitaxel chemoresistance in ovarian cancer cell lines. , 2009, Biochimica et biophysica acta.

[55]  Hongzhe Sun,et al.  Gold(III) porphyrins as a new class of anticancer drugs: cytotoxicity, DNA binding and induction of apoptosis in human cervix epitheloid cancer cells. , 2003, Chemical communications.

[56]  N. Karp,et al.  Design and Analysis Issues in Quantitative Proteomics Studies , 2007, Proteomics.

[57]  Qing‐Yu He,et al.  Proteomic characterization of the cytotoxic mechanism of gold (III) porphyrin 1a, a potential anticancer drug , 2006, Proteomics.

[58]  Brad T. Sherman,et al.  Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources , 2008, Nature Protocols.

[59]  K. Greulich,et al.  Genes of glycolysis are ubiquitously overexpressed in 24 cancer classes. , 2004, Genomics.

[60]  Dong Wang,et al.  Cellular processing of platinum anticancer drugs , 2005, Nature Reviews Drug Discovery.

[61]  A. Casini,et al.  Thioredoxin reductase: A target for gold compounds acting as potential anticancer drugs , 2009 .

[62]  D. Sawyer,et al.  Heat shock proteins in cancer: chaperones of tumorigenesis. , 2006, Trends in biochemical sciences.

[63]  A. Casini,et al.  Chemistry, antiproliferative properties, tumor selectivity, and molecular mechanisms of novel gold(III) compounds for cancer treatment: a systematic study , 2009, JBIC Journal of Biological Inorganic Chemistry.

[64]  R. Barnhardt,et al.  Inspiratory-modulated neurons of the rostrolateral pons: effects of pulmonary afferent input , 1989, Brain Research.

[65]  P. Greengard,et al.  Small‐molecule inducers of Aβ‐42 peptide production share a common mechanism of action , 2012, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[66]  E. Tiekink.  Gold derivatives for the treatment of cancer. , 2002, Critical reviews in oncology/hematology.

[67]  D F Hochstrasser,et al.  Methods for increasing the resolution of two-dimensional protein electrophoresis. , 1988, Analytical biochemistry.

[68]  L. Messori,et al.  Mechanisms of cytotoxicity of selected organogold(III) compounds. , 2005, Journal of medicinal chemistry.

[69]  Wei Sun,et al.  Proteome Analysis of Hepatocellular Carcinoma by Two-dimensional Difference Gel Electrophoresis , 2007, Molecular & Cellular Proteomics.

[70]  Alexey V. Antonov,et al.  BioProfiling.de: analytical web portal for high-throughput cell biology , 2011, Nucleic Acids Res..

[71]  Yi Yao,et al.  Comparative proteomic analysis of colon cancer cells in response to oxaliplatin treatment. , 2009, Biochimica et biophysica acta.

[72]  Pier Giorgio Righetti,et al.  Blue silver: A very sensitive colloidal Coomassie G‐250 staining for proteome analysis , 2004, Electrophoresis.