Identification of androgen-coregulated protein networks from the microsomes of human prostate cancer cells

BackgroundAndrogens play a critical role in the development of prostate cancer-dysregulation of androgen-regulated growth pathways can led to hormone-refractory prostate cancer. A comprehensive understanding of androgen-regulated cellular processes has not been achieved to date. To this end, we have applied a large-scale proteomic approach to define cellular processes that are responsive to androgen treatment in LNCaP prostate cancer cells.ResultsUsing isotope-coded affinity tags and mass spectrometry we identified and quantified the relative abundance levels of 1,064 proteins and found that distinct cellular processes were coregulated by androgen while others were essentially unaffected. Subsequent pharmacological perturbation of the cellular process for energy generation confirmed that androgen starvation had a profound effect on this pathway.ConclusionsOur results provide evidence for the role of androgenic hormones in coordinating the expression of critical components involved in distinct cellular processes and further establish a foundation for the comprehensive reconstruction of androgen-regulated protein networks and pathways in prostate cancer cells.

[1]  G. Church,et al.  Finding DNA regulatory motifs within unaligned noncoding sequences clustered by whole-genome mRNA quantitation , 1998, Nature Biotechnology.

[2]  J. Casida,et al.  Metabolism of Rotenone in vitro by Tissue Homogenates from Mammals and Insects , 1967, Science.

[3]  S. Ferdinandusse,et al.  Subcellular localization and physiological role of alpha-methylacyl-CoA racemase. , 2000, Journal of lipid research.

[4]  B. Neumcke.  The action of uncouplers on lipid bilayer membranes. , 1975, Membranes.

[5]  D. Longo,et al.  Linking β-Catenin to Androgen-signaling Pathway* , 2002, The Journal of Biological Chemistry.

[6]  J. G. Patton,et al.  An RNA recognition motif (RRM) is required for the localization of PTB-associated splicing factor (PSF) to subnuclear speckles. , 2001, Experimental cell research.

[7]  J. Vega,et al.  Neuroendocrine cells in benign prostatic hyperplasia and prostatic carcinoma: effect of hormonal treatment. , 1997, Urologia internationalis.

[8]  A. Shabsigh,et al.  Transdifferentiation of prostate cancer cells to a neuroendocrine cell phenotype in vitro and in vivo. , 1999, The Journal of urology.

[9]  D. Botstein,et al.  Cluster analysis and display of genome-wide expression patterns. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[10]  S. Gygi,et al.  Quantitative analysis of complex protein mixtures using isotope-coded affinity tags , 1999, Nature Biotechnology.

[11]  M. Rubin,et al.  Alpha-Methylacyl-CoA Racemase: A Novel Tumor Marker Over-expressed in Several Human Cancers and Their Precursor Lesions , 2002, The American journal of surgical pathology.

[12]  L. Hood,et al.  Prostate-localized and androgen-regulated expression of the membrane-bound serine protease TMPRSS2. , 1999, Cancer research.

[13]  Y. Nakakita,et al.  Isolation of oligomycin A as a result of screening for antagonists of lipids. , 1980, The Journal of antibiotics.

[14]  H. Lilja,et al.  Partial characterization of a thyroid‐stimulating hormone‐like peptide in neuroendocrine cells of the human prostate gland , 1989, The Prostate.

[15]  S. J. Higgins,et al.  The endocrinology and developmental biology of the prostate. , 1987, Endocrine reviews.

[16]  H. Klocker,et al.  Mutant androgen receptor detected in an advanced-stage prostatic carcinoma is activated by adrenal androgens and progesterone. , 1993, Molecular endocrinology.

[17]  J. Trent,et al.  Alpha-methylacyl-CoA racemase: a new molecular marker for prostate cancer. , 2002, Cancer research.

[18]  D. Black,et al.  Alternative RNA splicing in the nervous system , 2001, Progress in Neurobiology.

[19]  S. Balk,et al.  Androgen receptor as a target in androgen-independent prostate cancer. , 2002, Urology.

[20]  G. Bubley,et al.  Mutation of the androgen-receptor gene in metastatic androgen-independent prostate cancer. , 1995, The New England journal of medicine.

[21]  Bert W O'Malley,et al.  Coordinate Regulation of Transcription and Splicing by Steroid Receptor Coregulators , 2002, Science.

[22]  W. Birchmeier,et al.  Tumor-suppressor gene products in cell contacts: the cadherin-APC-armadillo connection. , 1994, Current opinion in cell biology.

[23]  J. Eastham,et al.  Androgen receptor mutations in prostate cancer. , 2000, Cancer research.

[24]  O. Lukkarinen,et al.  Mutated human androgen receptor gene detected in a prostatic cancer patient is also activated by estradiol. , 1995, The Journal of clinical endocrinology and metabolism.

[25]  E. Gelmann,et al.  Molecular biology of the androgen receptor. , 2002, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[26]  International Human Genome Sequencing Consortium.  Initial sequencing and analysis of the human genome , 2001, Nature.

[27]  R. Hubert,et al.  Catalytic cleavage of the androgen-regulated TMPRSS2 protease results in its secretion by prostate and prostate cancer epithelia. , 2001, Cancer research.

[28]  Debashis Ghosh,et al.  alpha-Methylacyl coenzyme A racemase as a tissue biomarker for prostate cancer. , 2002, JAMA.

[29]  Takashi Suda,et al.  Molecular cloning and expression of the fas ligand, a novel member of the tumor necrosis factor family , 1993, Cell.

[30]  Robert Tibshirani,et al.  Transcriptional programs activated by exposure of human prostate cancer cells to androgen , 2002, Genome Biology.

[31]  Michael Ashburner,et al.  On ontologies for biologists: the Gene Ontology--untangling the web. , 2002, Novartis Foundation symposium.

[32]  C. Huggins,et al.  Studies on prostatic cancer: I. The effect of castration, of estrogen and of androgen injection on serum phosphatases in metastatic carcinoma of the prostate , 1941, CA: a cancer journal for clinicians.

[33]  A. Gomez-Muñoz,et al.  Rapid activation of glycogen phosphorylase by steroid hormones in cultured rat hepatocytes. , 1989, The Biochemical journal.

[34]  R. Ian Freshney,et al.  Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications , 2010 .

[35]  R. Franklin,et al.  The Intermediary Metabolism of the Prostate: A Key to Understanding the Pathogenesis and Progression of Prostate Malignancy , 2000, Oncology.

[36]  E. Small,et al.  Selection for androgen receptor mutations in prostate cancers treated with androgen antagonist. , 1999, Cancer research.

[37]  P. Brazhnik,et al.  Gene networks: how to put the function in genomics. , 2002, Trends in biotechnology.

[38]  E. Wilson,et al.  Liganded androgen receptor interaction with beta-catenin: nuclear co-localization and modulation of transcriptional activity in neuronal cells. , 2002, The Journal of biological chemistry.

[39]  S. Reed,et al.  P504S: A New Molecular Marker for the Detection of Prostate Carcinoma , 2001, The American journal of surgical pathology.

[40]  M. Schober,et al.  Identification of differentially expressed genes by serial analysis of gene expression in human prostate cancer. , 2001, Cancer research.

[41]  C. Huggins,et al.  Studies on prostatic cancer: I. The effect of castration, of estrogen and of androgen injection on serum phosphatases in metastatic carcinoma of the prostate. 1941. , 2002, The Journal of urology.

[42]  Hanami Takeshi.  [Molecular probes]. , 2004, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme.

[43]  W. Isaacs,et al.  Androgen receptor gene mutations in human prostate cancer. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[44]  J. Swinnen,et al.  Androgens stimulate fatty acid synthase in the human prostate cancer cell line LNCaP. , 1997, Cancer research.

[45]  S. Hanash,et al.  Identification of androgen‐regulated genes in the prostate cancer cell line LNCaP by serial analysis of gene expression and proteomic analysis , 2001, Proteomics.

[46]  G. Azzone,et al.  Effect of funiculosin and antimycin A on the redox-driven H+-pumps in mitochondria: on the nature of "leaks'. , 1981, European journal of biochemistry.

[47]  R. Aebersold,et al.  Quantitative profiling of differentiation-induced microsomal proteins using isotope-coded affinity tags and mass spectrometry , 2001, Nature Biotechnology.

[48]  J. Trent,et al.  α-methylacyl-CoA racemase: A new molecular marker for prostate cancer , 2002 .

[49]  E. Gelmann,et al.  β-Catenin Binds to the Activation Function 2 Region of the Androgen Receptor and Modulates the Effects of the N-Terminal Domain and TIF2 on Ligand-Dependent Transcription , 2003, Molecular and Cellular Biology.

[50]  Biaoyang Lin,et al.  The program of androgen-responsive genes in neoplastic prostate epithelium , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[51]  Alexey I Nesvizhskii,et al.  Empirical statistical model to estimate the accuracy of peptide identifications made by MS/MS and database search. , 2002, Analytical chemistry.

[52]  S. Gygi,et al.  Correlation between Protein and mRNA Abundance in Yeast , 1999, Molecular and Cellular Biology.

[53]  J. Blenis,et al.  Cloning and Characterization of a Human STE20-like Protein Kinase with Unusual Cofactor Requirements* , 1997, The Journal of Biological Chemistry.

[54]  R. Schüle,et al.  FHL2, a novel tissue‐specific coactivator of the androgen receptor , 2000, The EMBO journal.

[55]  J. Swinnen,et al.  Coordinate regulation of lipogenic gene expression by androgens: evidence for a cascade mechanism involving sterol regulatory element binding proteins. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[56]  Trey Ideker,et al.  Transcriptome profiling to identify genes involved in peroxisome assembly and function , 2002, The Journal of cell biology.