Proteomic analysis of MCF-7 cells treated with benzo[a]pyrene, dibenzo[a,l]pyrene, coal tar extract, and diesel exhaust extract.

Polycyclic aromatic hydrocarbon (PAH) DNA adducts have been associated with carcinogenesis, which is accompanied by multiple alterations in gene expression. We used two-dimensional electrophoresis to distinguish protein expression changes induced in MCF-7 cells by individual PAH (B[a]P and DB[a,l]P) and PAH mixtures (coal tar extract [SRM 1597] and diesel exhaust extract [SRM 1975]). Spots of interest were identified by MALDI-TOF-TOF. Our results have shown alterations in the expression of heat-shock proteins, cytoskeletal proteins, DNA associated proteins, and glycolytic and mitochondrial proteins. The proteins that were universally altered in expression were actin cytoplasmic 1, tubulin alpha and myosin light chain alkali, cyclophilin B, and heterogeneous ribonucleoprotein B1 (a protein involved in access to telomerase and mRNA maturation). Additional proteins with altered expression include histone H2A.1, heat-shock protein 70-2, galectin-3, nucleoside diphosphate kinase, ATP synthase, and electron transfer flavoprotein. While sharing similarities, each PAH treatment exhibited a unique proteomic fingerprint.

[1]  R. Smart,et al.  A multihit, multistage model of chemical carcinogenesis. , 1999, Carcinogenesis.

[2]  I. Weinstein,et al.  Interaction of benzo(a)pyrene and its dihydrodiol-epoxide derivative with nuclear and mitochondrial DNA in C3H10T 1/2 cell cultures. , 1982, Cancer research.

[3]  L. McCarty,et al.  Can mode of action predict mixture toxicity for risk assessment? , 2004, Toxicology and applied pharmacology.

[4]  J. Emery,et al.  Unique gene expression patterns in liver and kidney associated with exposure to chemical toxicants. , 2001, The Journal of pharmacology and experimental therapeutics.

[5]  J. T. Wachsman,et al.  Mitochondrial DNA in human malignancy. , 2001, Mutation research.

[6]  K. Miller,et al.  IMPACT OF CELLULAR METABOLISM ON THE BIOLOGICAL EFFECTS OF BENZO[A]PYRENE AND RELATED HYDROCARBONS† , 2001, Drug metabolism reviews.

[7]  M. Jäättelä,et al.  The heat shock protein 70 family: Highly homologous proteins with overlapping and distinct functions , 2007, FEBS letters.

[8]  B. Mahadevan,et al.  Altered gene expression patterns in MCF-7 cells induced by the urban dust particulate complex mixture standard reference material 1649a. , 2005, Cancer research.

[9]  F. Larcher,et al.  Aberrant expression of the simple epithelial type II keratin 8 by mouse skin carcinomas but not papillomas , 1992, Molecular carcinogenesis.

[10]  Chi-Ying F. Huang,et al.  Benzo[a]pyrene and glycine N-methyltransferse interactions: gene expression profiles of the liver detoxification pathway. , 2006, Toxicology and applied pharmacology.

[11]  L. Ehrenberg,et al.  Relevance of different biological assays in assessing initiating and promoting properties of polycyclic aromatic hydrocarbons with respect to carcinogenic potency. , 1996, Mutation research.

[12]  A. Luch,et al.  Cytochrome P450 1B1 determines susceptibility to dibenzo[a,l]pyrene-induced tumor formation. , 2002, Chemical research in toxicology.

[13]  G. Perdew,et al.  A dynamic role for the Ah receptor in cell signaling? Insights from a diverse group of Ah receptor interacting proteins , 2002, Journal of biochemical and molecular toxicology.

[14]  B. Mahadevan,et al.  Carcinogenic polycyclic aromatic hydrocarbon‐DNA adducts and mechanism of action , 2005, Environmental and molecular mutagenesis.

[15]  I. Giddings,et al.  Time- and concentration-dependent changes in gene expression induced by benzo(a)pyrene in two human cell lines, MCF-7 and HepG2 , 2006, BMC Genomics.

[16]  M. Tang,et al.  Targeting of lung cancer mutational hotspots by polycyclic aromatic hydrocarbons. , 2000, Journal of the National Cancer Institute.

[17]  W. Cho,et al.  Contribution of oncoproteomics to cancer biomarker discovery , 2007, Molecular Cancer.

[18]  E. Friederich,et al.  The actin cytoskeleton as a therapeutic target: state of the art and future directions. , 2003, Progress in cell cycle research.

[19]  M A Sirover,et al.  New insights into an old protein: the functional diversity of mammalian glyceraldehyde-3-phosphate dehydrogenase. , 1999, Biochimica et biophysica acta.

[20]  Thomas J. Slavin,et al.  A Critical Assessment of Studies on the Carcinogenic Potential of Diesel Exhaust , 2006, Critical reviews in toxicology.

[21]  J. Ferguson,et al.  Effect of a complex environmental mixture from coal tar containing polycyclic aromatic hydrocarbons (PAH) on the tumor initiation, PAH-DNA binding and metabolic activation of carcinogenic PAH in mouse epidermis. , 2001, Carcinogenesis.

[22]  R. Harvey,et al.  Polycyclic Aromatic Hydrocarbons: Chemistry and Carcinogenicity , 1992 .

[23]  P. Cullen,et al.  The Ras binary switch: an ideal processor for decoding complex Ca2+ signals? , 2003, Biochemical Society transactions.

[24]  A. Sakurada,et al.  hnRNP B1 protein may be a possible prognostic factor in squamous cell carcinoma of the lung. , 2003, Lung cancer.

[25]  L. Hendershot,et al.  A subset of chaperones and folding enzymes form multiprotein complexes in endoplasmic reticulum to bind nascent proteins. , 2002, Molecular biology of the cell.

[26]  Q. Khan,et al.  Hydrocarbon carcinogens evade cellular defense mechanism of G1 arrest in nontransformed and malignant lung cell lines. , 2001, Toxicology and applied pharmacology.

[27]  P Boffetta,et al.  Occupational exposures to polycyclic aromatic hydrocarbons, and respiratory and urinary tract cancers: a quantitative review to 2005. , 2006, Annals of oncology : official journal of the European Society for Medical Oncology.

[28]  L. Weiss,et al.  Cytokeratin 14 expression in epithelial neoplasms: a survey of 435 cases with emphasis on its value in differentiating squamous cell carcinomas from other epithelial tumours , 2001, Histopathology.

[29]  P. Kovacic,et al.  Mechanisms of carcinogenesis: focus on oxidative stress and electron transfer. , 2001, Current medicinal chemistry.

[30]  H. Zhu,et al.  Characterization of benzo[a]pyrene quinone-induced toxicity to primary cultured bone marrow stromal cells from DBA/2 mice: potential role of mitochondrial dysfunction. , 1995, Toxicology and applied pharmacology.

[31]  D. Benjamin,et al.  mRNA stability and cancer: an emerging link? , 2007, Expert opinion on biological therapy.

[32]  F Kalberlah,et al.  Cancer risk assessment for oral exposure to PAH mixtures , 2002, Journal of applied toxicology : JAT.

[33]  Oliana Carnevali,et al.  Exposure to xenobiotic compounds: looking for new biomarkers. , 2003, General and comparative endocrinology.

[34]  D. N. Perkins,et al.  Probability‐based protein identification by searching sequence databases using mass spectrometry data , 1999, Electrophoresis.

[35]  John Condeelis,et al.  The cofilin pathway in breast cancer invasion and metastasis , 2007, Nature Reviews Cancer.

[36]  D. Phillips,et al.  Covalent binding of dibenzpyrenes and benzo[a]pyrene to DNA: evidence for synergistic and inhibitory interactions when applied in combination to mouse skin. , 1990, Carcinogenesis.

[37]  A. Dipple,et al.  Polycyclic aromatic hydrocarbon carcinogens. , 1990, Progress in clinical and biological research.

[38]  R. Finley,et al.  Galectin-3 Translocates to the Perinuclear Membranes and Inhibits Cytochrome c Release from the Mitochondria , 2002, The Journal of Biological Chemistry.

[39]  M. Rentrop,et al.  Aberrant expression during two‐stage mouse skin carcinogenesis of a type 147‐kDa keratin, k13, normally associated with terminal differentiation of internal stratified epithelia , 1988, Molecular carcinogenesis.

[40]  A. Dipple,et al.  7,12-dimethylbenz[a] anthracene--DNA binding in mouse skin: response of different mouse strains and effects of various modifiers of carcinogenesis. , 1984, Carcinogenesis.

[41]  P. Percipalle,et al.  The function of actin in gene transcription. , 2007, Histology and histopathology.

[42]  A. Luch,et al.  Stereoselective activation of dibenzo[a,l]pyrene and its trans-11,12-dihydrodiol to fjord region 11,12-diol 13,14-epoxides in a human mammary carcinoma MCF-7 cell-mediated V79 cell mutation assay. , 1997, Chemical research in toxicology.

[43]  S. Safe,et al.  Characterization of calcium oscillations in normal and benzo[a]pyrene-treated clone 9 cells. , 2002, Toxicological sciences : an official journal of the Society of Toxicology.

[44]  W. Baird,et al.  Diesel exhaust influences carcinogenic PAH-induced genotoxicity and gene expression in human breast epithelial cells in culture. , 2007, Mutation research.

[45]  C. B. Pickett,et al.  Regulatory mechanisms controlling gene expression mediated by the antioxidant response element. , 2003, Annual review of pharmacology and toxicology.

[46]  Stairs Pw,et al.  Benzo[a]pyrene differentially alters mitochondrial and nuclear DNA synthesis in primary hepatocyte cultures. , 1983 .

[47]  N. Galeva,et al.  Comparison of one‐dimensional and two‐dimensional gel electrophoresis as a separation tool for proteomic analysis of rat liver microsomes: Cytochromes P450 and other membrane proteins , 2002, Proteomics.

[48]  Lue Ping Zhao,et al.  Phenotypic anchoring of global gene expression profiles induced by N-hydroxy-4-acetylaminobiphenyl and benzo[a]pyrene diol epoxide reveals correlations between expression profiles and mechanism of toxicity. , 2005, Chemical research in toxicology.

[49]  Yu-Min Shen,et al.  Comparison of p53 mutations induced by PAH o-quinones with those caused by anti-benzo[a]pyrene diol epoxide in vitro: role of reactive oxygen and biological selection. , 2006, Chemical research in toxicology.

[50]  David M DeMarini,et al.  Transcriptional responses to complex mixtures: a review. , 2007, Mutation research.

[51]  A. Weston,et al.  Transcriptional signatures of environmentally relevant exposures in normal human mammary epithelial cells: benzo[a]pyrene. , 2005, Cancer letters.

[52]  H. Satoh,et al.  Expression of hnRNP A2/B1 proteins in human cancer cell lines. , 2000, International journal of oncology.

[53]  W. Baird,et al.  Benzo(e)pyrene-induced alterations in the metabolic activation of benzo(a)pyrene and 7,12-dimethylbenz(a)anthracene by hamster embryo cells. , 1984, Cancer research.

[54]  William C Copeland,et al.  Mitochondrial DNA Alterations in Cancer , 2002, Cancer investigation.

[55]  T. Fujimori,et al.  RT-PCR amplification of RNA extracted from formalin-fixed, paraffin-embedded oral cancer sections: analysis of p53 pathway. , 2003, Anticancer research.

[56]  T. Slaga,et al.  Early expression of type I K13 keratin in the progression of mouse skin papillomas. , 1990, Carcinogenesis.

[57]  M. Fukuda,et al.  Nucleoside Diphosphate Kinases in Mammalian Signal Transduction Systems: Recent Development and Perspective , 2003, Journal of bioenergetics and biomembranes.

[58]  R D Appel,et al.  Protein identification and analysis tools in the ExPASy server. , 1999, Methods in molecular biology.

[59]  R. Tukey,et al.  The Role of the Ah Receptor and p38 in Benzo[a]pyrene-7,8-dihydrodiol and Benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide-induced Apoptosis* , 2003, Journal of Biological Chemistry.

[60]  D. Aden,et al.  Histone complements of human tissues, carcinomas, and carcinoma-derived cell lines , 1984, Molecular and Cellular Biochemistry.

[61]  C. Qiu,et al.  Gene expression profiling in the mammary gland of rats treated with 7,12‐dimethylbenz[a]anthracene , 2006, International journal of cancer.

[62]  D. Hebels,et al.  Binary PAH mixtures cause additive or antagonistic effects on gene expression but synergistic effects on DNA adduct formation. , 2007, Carcinogenesis.

[63]  G. Semenza,et al.  'The metabolism of tumours': 70 years later. , 2001, Novartis Foundation symposium.

[64]  W. A. Toscano,et al.  Transcriptional regulation of glyceraldehyde-3-phosphate dehydrogenase by 2,3,7,8-tetrachlorodibenzo-p-dioxin. , 1995, Biochemical and biophysical research communications.

[65]  C. Heidelberger,et al.  Binding of tritium-labeled polycyclic hydrocarbons to DNA of mouse skin. , 1967, Cancer research.

[66]  H. Rubin.  Synergistic mechanisms in carcinogenesis by polycyclic aromatic hydrocarbons and by tobacco smoke: a bio-historical perspective with updates. , 2001, Carcinogenesis.

[67]  R. Somogyi,et al.  Gene Expression Microarray Data Analysis for Toxicology Profiling , 2000, Annals of the New York Academy of Sciences.

[68]  J. Trosko,et al.  The Emperor Wears No Clothes in the Field of Carcinogen Risk Assessment: Ignored Concepts in Cancer Risk Assessment , 2005 .

[69]  S M Morris,et al.  Gene expression profiles and genetic damage in benzo(a)pyrene diol epoxide-exposed TK6 cells. , 2004, Mutation research.

[70]  D. Phillips,et al.  32P-postlabelling analysis of the covalent binding of benzo[ghi]perylene to DNA in vivo and in vitro. , 1993, Carcinogenesis.

[71]  F. Oswald,et al.  The E2F transcription factor activates a replication-dependent human H2A gene in early S phase of the cell cycle , 1996, Molecular and cellular biology.

[72]  V. Pancholi,et al.  Multifunctional α-enolase: its role in diseases , 2001, Cellular and Molecular Life Sciences CMLS.

[73]  S. Ohta.  A multi-functional organelle mitochondrion is involved in cell death, proliferation and disease. , 2003, Current medicinal chemistry.

[74]  P. Hainaut,et al.  Patterns of p53 G-->T transversions in lung cancers reflect the primary mutagenic signature of DNA-damage by tobacco smoke. , 2001, Carcinogenesis.

[75]  G. Kroemer,et al.  HSP27 and HSP70: Potentially Oncogenic Apoptosis Inhibitors , 2003, Cell cycle.

[76]  W. Au,et al.  Variant metabolizing gene alleles determine the genotoxicity of benzo[a]pyrene , 2001, Environmental and molecular mutagenesis.

[77]  N. Robinson,et al.  Application of Benzo(a)pyrene and Coal Tar Tumor Dose–Response Data to a Modified Benchmark Dose Method of Guideline Development , 2004, Environmental health perspectives.

[78]  J. Going,et al.  Stages on the way to breast cancer , 2003, The Journal of pathology.

[79]  S. Hsu,et al.  Benzo[g,h,i]perylene synergistically transactivates benzo[a]pyrene-induced CYP1A1 gene expression by aryl hydrocarbon receptor pathway. , 2001, Toxicology and applied pharmacology.

[80]  E. Sausville,et al.  Aryl hydrocarbon receptor mediates sensitivity of MCF-7 breast cancer cells to antitumor agent 2-(4-amino-3-methylphenyl) benzothiazole. , 2002, Molecular pharmacology.

[81]  C. Kahn,et al.  Regulation of growth and tumorigenicity of breast cancer cells by the low molecular weight GTPase Rad and nm23. , 2001, Cancer research.

[82]  John M. Walker,et al.  The Proteomics Protocols Handbook , 2005, Humana Press.

[83]  Q. Khan,et al.  DNA reactions, mutagenic action and stealth properties of polycyclic aromatic hydrocarbon carcinogens (review). , 1999, International journal of oncology.

[84]  I. Weinstein,et al.  Mitochondrial DNA is a major cellular target for a dihydrodiol-epoxide derivative of benzo[a]pyrene. , 1980, Science.

[85]  K. Fujimori,et al.  Enhanced expression of a glyceraldehyde-3-phosphate dehydrogenase gene in human lung cancers. , 1987, Cancer research.

[86]  V. Kobliakov,et al.  Effect of some carcinogenic and non-carcinogenic polycyclic aromatic hydrocarbons on gap junction intercellular communication in hepatoma cell cultures. , 2006, European journal of cell biology.

[87]  A. Long,et al.  A human cell line from a pleural effusion derived from a breast carcinoma. , 1973, Journal of the National Cancer Institute.

[88]  R. Anzion,et al.  Highly increased urinary 1-hydroxypyrene excretion rate in patients with atopic dermatitis treated with topical coal tar , 2002, Archives of Dermatological Research.

[89]  J. Whitlock,et al.  Induction of cytochrome P4501A1. , 1999, Annual review of pharmacology and toxicology.

[90]  Cathy H. Wu,et al.  The Universal Protein Resource (UniProt): an expanding universe of protein information , 2005, Nucleic Acids Res..