Molecular Mechanisms of Multistage Carcinogenesis

Summary

Principal Investigator: TREVOR PENNING
Abstract: DESCRIPTION (provided by applicant) Cancer is the second leading cause of death in the United States and involves multiple stages including, initiation, promotion, progression, and metastasis. This Program will dissect the discrete molecular events associated with early components of this multistage disease by using complete carcinogens as chemical tweezers [polycyclic aromatic hydrocarbons (PAH)]. The hypothesis to be tested is that reactive and redox active PAH o-quinones generated by constitutively expressed aldo-keto reductase (AKR?s)contribute to PAHmultistage carcinogenesis. We propose that the electrophilic PAH-o-quinones plus the reactive oxygen species (ROS) they generate lead to covalent modifications of DNA in PAH target tissues and this may have mutational consequences leading to initiation. We also propose that the PAH o-quinones and ROS have direct effects on protein kinase C and its downstream signals. These epigenetic effects may have consequences for cell proliferation leading to promotion. These events could explain how PAH act as complete carcinogens. The Program consists of three interactive Projects and two Cores. Each project will use common human cell lines which are either capable of PAH activation (MCF-7 cells) or are PAH targets (bronchoalveolar cells). Project # 1 (Dr. Penning) will compare the roles of human aldehyde reductase (AKR1A1), CYP1A1/CYP1B1, and CYP-peroxidase in the metabolic activation of PAH using a stable expression strategy. Project #2 (Dr. Blair) will develop quantitative LC/MS methods to measure covalent modifications to DNA by reactive PAH-metabolites (o-quinones, anti-diol epoxides and radical cations), ROS and decomposition products of lipid hydroperoxides. The relative abundance of these DNA-adducts will be measured in bronchoalveolar cells (Project #1) and in sites of PAH carcinogenesis i.e, SENCAR mouse skin (Project #2). This will identify the metabolic pathway most responsible for adduct formation. Project #3 (Drs. Kazanietz and Assoian) will determine whether PAH o-quinones and ROS activate or inhibit individual PKC isoforms in vitro and in vivo and measure the phenotypic outcome on PMA-induced cell proliferation or growth inhibition. Changes in growth properties will be correlated to effects on cell cycle signaling pathways. The Bioanalytical Core (Core B) will provide LC/MS support to monitor the purity of PAH metabolites used in all projects, and will provide authentic PAH-DNA adduct reference compounds for Projects 1 and 2. The administrative core (Core A) will provide oversight to the program, a share Program Database, and biostatistical support to tract trends within and between projects. By studying the discrete molecular events responsible for the causation of cancer this Program may lead to the early prevention and intervention of this disease.
Funding Period: 2002-08-30 - 2007-07-31
more information: NIH RePORT

Top Publications

  1. pmc Specificity of human aldo-keto reductases, NAD(P)H:quinone oxidoreductase, and carbonyl reductases to redox-cycle polycyclic aromatic hydrocarbon diones and 4-hydroxyequilenin-o-quinone
    Carol A Shultz
    Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA
    Chem Res Toxicol 24:2153-66. 2011
  2. ncbi Efficient syntheses of C(8)-aryl adducts of adenine and guanine formed by reaction of radical cation metabolites of carcinogenic polycyclic aromatic hydrocarbons with DNA
    Qing Dai
    The Ben May Department for Cancer Research, The University of Chicago, Chicago, Illinois 60637, USA
    J Org Chem 72:4856-63. 2007
  3. pmc Human aldo-keto reductases: Function, gene regulation, and single nucleotide polymorphisms
    Trevor M Penning
    Center of Excellence in Environmental Toxicology, Department of Pharmacology, University of Pennsylvania School of Medicine, 130 C John Morgan Bldg, 3620 Hamilton Walk, Philadelphia, PA 19104 6084, USA
    Arch Biochem Biophys 464:241-50. 2007
  4. pmc Metabolism of benzo[a]pyrene in human bronchoalveolar H358 cells using liquid chromatography-mass spectrometry
    Hao Jiang
    Centers of Excellence in Environmental Toxicology and Cancer Pharmacology, Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104 6084, USA
    Chem Res Toxicol 20:1331-41. 2007
  5. ncbi S-Phase-specific activation of PKC alpha induces senescence in non-small cell lung cancer cells
    Jose L Oliva
    Department of Pharmacology and Institute for Translational Medicine and Therapeutics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104 6160, USA
    J Biol Chem 283:5466-76. 2008
  6. ncbi Strategies for synthesis of adducts of omicron-quinone metabolites of carcinogenic polycyclic aromatic hydrocarbons with 2'-deoxyribonucleosides
    Chongzhao Ran
    The Ben May Department for Cancer Research, The University of Chicago, Chicago, Illinois 60637, USA
    J Org Chem 73:992-1003. 2008
  7. pmc Fjord-region benzo[g]chrysene-11,12-dihydrodiol and benzo[c]phenanthrene-3,4-dihydrodiol as substrates for rat liver dihydrodiol dehydrogenase (AKR1C9): structural basis for stereochemical preference
    Carol A Shultz
    Department of Biochemistry and Biophysics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA
    Chem Res Toxicol 21:668-77. 2008
  8. ncbi Synthesis of dibenzo[def,p]chrysene, its active metabolites, and their 13C-labeled analogues
    Daiwang Xu
    Ben May Department for Cancer Research, University of Chicago, Chicago, IL 60637, USA
    Org Lett 10:1059-62. 2008
  9. ncbi Comparisons of (+/-)-benzo[a]pyrene-trans-7,8-dihydrodiol activation by human cytochrome P450 and aldo-keto reductase enzymes: effect of redox state and expression levels
    Amy M Quinn
    Center of Excellence in Environmental Toxicology, Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104 6084, USA
    Chem Res Toxicol 21:1086-94. 2008
  10. pmc The pattern of p53 mutations caused by PAH o-quinones is driven by 8-oxo-dGuo formation while the spectrum of mutations is determined by biological selection for dominance
    Jong Heum Park
    Department of Pharmacology, Centers for Excellence in Environmental Toxicology and Cancer Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104 6084, USA
    Chem Res Toxicol 21:1039-49. 2008

Scientific Experts

  • TREVOR PENNING
  • Jose L Oliva
  • Amy M Quinn
  • Ronald G Harvey
  • Ian A Blair
  • Jong Heum Park
  • Hao Jiang
  • Qing Dai
  • Daiwang Xu
  • Qian Ruan
  • Chongzhao Ran
  • Carol A Shultz
  • Andrea B Troxel
  • Yu Min Shen
  • Dipti Mangal
  • Jeffrey Field
  • Srilakshmi Vedantam
  • Yi Jin
  • Stacy L Gelhaus
  • Janice L Walker
  • Marcelo G Kazanietz
  • Jun Gao
  • Ademi E Santiago-Walker
  • Edmund Maser
  • Judy L Bolton
  • Nisha T Palackal
  • Stacy Gelhaus
  • Andrea L Oliva
  • Yazhen Duan
  • Abhita Batra
  • Keunpoong Lim
  • Paola Castagnino
  • Richard K Assoian
  • Betty M Chung
  • Hye Young H Kim
  • Daljit K Vudathala
  • Gary D Kao
  • Lawrence M Szewczuk
  • Scott W Burchiel
  • Fredine T Lauer
  • Judy L Meinkoth
  • Andrew A Voss
  • Sridhar Gopishetty
  • Eric J Brown
  • Isaac N Pessah
  • Aphrothiti J Fikaris

Detail Information

Publications26

  1. pmc Specificity of human aldo-keto reductases, NAD(P)H:quinone oxidoreductase, and carbonyl reductases to redox-cycle polycyclic aromatic hydrocarbon diones and 4-hydroxyequilenin-o-quinone
    Carol A Shultz
    Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA
    Chem Res Toxicol 24:2153-66. 2011
    ..ROS formation was unaffected by the addition of dicumarol, suggesting that NQO1 is not responsible for the two-electron reduction observed and does not offer protection against ROS formation from PAH o-quinones...
  2. ncbi Efficient syntheses of C(8)-aryl adducts of adenine and guanine formed by reaction of radical cation metabolites of carcinogenic polycyclic aromatic hydrocarbons with DNA
    Qing Dai
    The Ben May Department for Cancer Research, The University of Chicago, Chicago, Illinois 60637, USA
    J Org Chem 72:4856-63. 2007
    ..anthracene, benz[a]anthracene, benzo[a]pyrene, and dibenzo[def,p]chrysene, could not be obtained. This difference was shown to be a consequence of the facility of competing hydrolytic deboronation of the corresponding arylboronic acids...
  3. pmc Human aldo-keto reductases: Function, gene regulation, and single nucleotide polymorphisms
    Trevor M Penning
    Center of Excellence in Environmental Toxicology, Department of Pharmacology, University of Pennsylvania School of Medicine, 130 C John Morgan Bldg, 3620 Hamilton Walk, Philadelphia, PA 19104 6084, USA
    Arch Biochem Biophys 464:241-50. 2007
    ..This suggests that there will be inter-individual variation in endogenous and xenobiotic metabolism which in turn affect susceptibility to nuclear receptor signaling and chemical carcinogenesis...
  4. pmc Metabolism of benzo[a]pyrene in human bronchoalveolar H358 cells using liquid chromatography-mass spectrometry
    Hao Jiang
    Centers of Excellence in Environmental Toxicology and Cancer Pharmacology, Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104 6084, USA
    Chem Res Toxicol 20:1331-41. 2007
    ..It also provides evidence that each of the pathways of PAH activation yields their distinctive metabolites in H358 human lung cells and that each pathway may contribute to the carcinogenic process...
  5. ncbi S-Phase-specific activation of PKC alpha induces senescence in non-small cell lung cancer cells
    Jose L Oliva
    Department of Pharmacology and Institute for Translational Medicine and Therapeutics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104 6160, USA
    J Biol Chem 283:5466-76. 2008
    ....
  6. ncbi Strategies for synthesis of adducts of omicron-quinone metabolites of carcinogenic polycyclic aromatic hydrocarbons with 2'-deoxyribonucleosides
    Chongzhao Ran
    The Ben May Department for Cancer Research, The University of Chicago, Chicago, Illinois 60637, USA
    J Org Chem 73:992-1003. 2008
    ..The copper-mediated method offers advantages of economy, adaptability to large-scale preparation, utility for synthesis of (13)C- or (15)N-labeled analogues, and nonformation of bis-adducts as secondary products...
  7. pmc Fjord-region benzo[g]chrysene-11,12-dihydrodiol and benzo[c]phenanthrene-3,4-dihydrodiol as substrates for rat liver dihydrodiol dehydrogenase (AKR1C9): structural basis for stereochemical preference
    Carol A Shultz
    Department of Biochemistry and Biophysics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA
    Chem Res Toxicol 21:668-77. 2008
    ..The similarity between rates of trans-dihydrodiol oxidation by the rat and human liver specific AKRs (AKR1C9 and AKR1C4) implicate these enzymes in hepatocarcinogenesis in rats observed with the fjord-region PAH...
  8. ncbi Synthesis of dibenzo[def,p]chrysene, its active metabolites, and their 13C-labeled analogues
    Daiwang Xu
    Ben May Department for Cancer Research, University of Chicago, Chicago, IL 60637, USA
    Org Lett 10:1059-62. 2008
    ..The 13C2-labeled analogues are required as standards for sensitive methods of analysis of their DNA adducts in human cells using stable isotope dilution liquid chromatography/tandem mass spectrometry...
  9. ncbi Comparisons of (+/-)-benzo[a]pyrene-trans-7,8-dihydrodiol activation by human cytochrome P450 and aldo-keto reductase enzymes: effect of redox state and expression levels
    Amy M Quinn
    Center of Excellence in Environmental Toxicology, Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104 6084, USA
    Chem Res Toxicol 21:1086-94. 2008
    ..These data suggest that AKR enzymes may effectively compete with P450 1A1/1B1 for PAH trans-dihydrodiol activation in human lung cells...
  10. pmc The pattern of p53 mutations caused by PAH o-quinones is driven by 8-oxo-dGuo formation while the spectrum of mutations is determined by biological selection for dominance
    Jong Heum Park
    Department of Pharmacology, Centers for Excellence in Environmental Toxicology and Cancer Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104 6084, USA
    Chem Res Toxicol 21:1039-49. 2008
    ..These observations, if extended to mammalian cells, suggest that mutagenesis can drive the pattern of mutations but that biological selection for dominant mutations drives the spectrum of mutations observed in p53 in lung cancer...
  11. pmc Synthesis of phenol and quinone metabolites of benzo[a]pyrene, a carcinogenic component of tobacco smoke implicated in lung cancer
    Daiwang Xu
    The Ben May Department for Cancer Research, The University of Chicago, Chicago, Illinois 60637, USA
    J Org Chem 74:597-604. 2009
    ....
  12. ncbi Aldo-keto reductase- and cytochrome P450-dependent formation of benzo[a]pyrene-derived DNA adducts in human bronchoalveolar cells
    Qian Ruan
    Centers for Cancer Pharmacology and Excellence in Environmental Toxicology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104 6160, USA
    Chem Res Toxicol 20:424-31. 2007
    ..These data raise the intriguing possibility that P450 1A1/P450 1B1 and AKR1A1 may be protective against (+)-B[a]PDE-mediated DNA damage...
  13. pmc 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
    Yu Min Shen
    Department of Pharmacology, Center of Excellence in Environmental Toxicology and Center for Clinical Epidemiology and Biostatistics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104 6084, USA
    Chem Res Toxicol 19:1441-50. 2006
    ..The resultant 8-oxo-dGuo yields a pattern of mutations but not a spectrum consistent with that seen in lung cancer; we suggest that the emergence of the spectrum requires biological selection...
  14. ncbi AKR1B10: a new diagnostic marker of non-small cell lung carcinoma in smokers
    Trevor M Penning
    Department of Pharmacology, University of Pennsylvania School of Medicine, 3620 Hamilton Walk, Philadelphia, PA 19104 6084, USA
    Clin Cancer Res 11:1687-90. 2005
  15. ncbi Synthesis of adducts of o-quinone metabolites of carcinogenic polycyclic aromatic hydrocarbons with 2'-deoxyribonucleosides
    Qing Dai
    Ben May Institute for Cancer Research, The University of Chicago, Chicago, Illinois 60637, USA
    Org Lett 7:999-1002. 2005
    ..These syntheses entail Pd-catalyzed coupling of protected amine derivatives of catechols with suitably protected halopurine analogues of 2'-deoxyribonucleosides...
  16. ncbi N6-Arylation of 2'-deoxyadenosine via copper-catalyzed direct coupling with aryl halides
    Chongzhao Ran
    Ben May Institute for Cancer Research, University of Chicago, Chicago, Illinois 60637, USA
    J Org Chem 70:3724-6. 2005
    ..The method is useful for aryl halides with either electron-donating or electron-withdrawing groups...
  17. pmc Formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dGuo) by PAH o-quinones: involvement of reactive oxygen species and copper(II)/copper(I) redox cycling
    Jong Heum Park
    Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA
    Chem Res Toxicol 18:1026-37. 2005
    ....
  18. ncbi Ryanodine receptor-mediated rapid increase in intracellular calcium induced by 7,8-benzo(a)pyrene quinone in human and murine leukocytes
    Jun Gao
    The University of New Mexico College of Pharmacy Toxicology Program, Albuquerque, 87131 0001, USA
    Toxicol Sci 87:419-26. 2005
    ..Collectively, these results demonstrate a novel mechanism of Ca(2+) elevation by an environmentally relevant metabolite of BaP in murine and human lymphocytes...
  19. ncbi Protein kinase C delta stimulates apoptosis by initiating G1 phase cell cycle progression and S phase arrest
    Ademi E Santiago-Walker
    Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, 19104 6061, USA
    J Biol Chem 280:32107-14. 2005
    ..Importantly, they reveal the existence of a novel, cell cycle-dependent mechanism through which PKCdelta stimulates cell death...
  20. ncbi Competing roles of aldo-keto reductase 1A1 and cytochrome P4501B1 in benzo[a]pyrene-7,8-diol activation in human bronchoalveolar H358 cells: role of AKRs in P4501B1 induction
    Hao Jiang
    Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104 6084, USA
    Chem Res Toxicol 19:68-78. 2006
    ....
  21. ncbi Quantification of benzo[a]pyrene diol epoxide DNA-adducts by stable isotope dilution liquid chromatography/tandem mass spectrometry
    Qian Ruan
    Center for Cancer Pharmacology, University of Pennsylvania School of Medicine, 854 BRB II III, 421 Curie Boulevard, Philadelphia, PA 19104 6160, USA
    Rapid Commun Mass Spectrom 20:1369-80. 2006
    ..Also, the adduct profile suggests that this occurs by binding of (+)-anti-B[a]PDE to DNA in a manner that facilitates covalent binding to dGuo rather than dAdo residues...
  22. pmc Polycyclic aromatic hydrocarbon (PAH) o-quinones produced by the aldo-keto-reductases (AKRs) generate abasic sites, oxidized pyrimidines, and 8-oxo-dGuo via reactive oxygen species
    Jong Heum Park
    Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104 6084, USA
    Chem Res Toxicol 19:719-28. 2006
    ..However, in the presence of Fe(III)-mediated PAH o-quinone redox-cycling, the *OH radical scavengers and sodium azide consistently attenuated their formation, indicating that the ROS responsible was *OH...
  23. ncbi Aldo-keto reductases and bioactivation/detoxication
    Yi Jin
    Department of Pharmacology, Center of Excellence in Environmental Toxicology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA
    Annu Rev Pharmacol Toxicol 47:263-92. 2007
    ..g., aflatoxin, endogenous toxicants, and those formed from the breakdown of lipid peroxides. AKRs are stress-regulated genes and play a central role in the cellular response to osmotic, electrophilic, and oxidative stress...
  24. ncbi Post-transcriptional destabilization of p21cip1 by protein kinase C in fibroblasts
    Janice L Walker
    Department of Pharmacology, University of Pennsylvania School of Medicine, 3620 Hamilton Walk, Philadelphia, PA 19104 6084, USA
    J Biol Chem 281:38127-32. 2006
    ..Thus, post-transcriptional destabilization of p21(cip1) appears to be a major mitogenic effect of PKCdelta in fibroblasts...
  25. ncbi Competing roles of cytochrome P450 1A1/1B1 and aldo-keto reductase 1A1 in the metabolic activation of (+/-)-7,8-dihydroxy-7,8-dihydro-benzo[a]pyrene in human bronchoalveolar cell extracts
    Hao Jiang
    Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104 6084, USA
    Chem Res Toxicol 18:365-74. 2005
    ..These model systems provide a cellular context in which the dominant DNA adducts/lesions formed by either pathway may be compared...