Response of detoxification gene mRNA expression and selection of molecular biomarkers in the clam Ruditapes philippinarum exposed to benzo[a]pyrene.

Benzo[a]pyrene (B[a]P) has a high carcinogenic potential. B[a]P concentrations and molecular biomarkers (mRNA expressions of Pgp, AhR, CYP4, CYP414A1, GST-pi, GST-S2, Cu/Zn-SOD and Mn-SOD) were assayed in gills and digestive glands of the clam Ruditapes philippinarum exposed to 0.03, 0.3 and 3 μg/L B[a]P for 21 days and then exposed to natural seawater for 15 days. Results showed that B[a]P was rapidly accumulated in and then eliminated from tissues of the clams. All gene mRNA expressions in the treated groups were induced significantly with the exception of CYP414A1 and Cu/Zn-SOD in the 0.03 μg/L B[a]P group. According to correlation analysis, mRNA expressions of AhR, GST-pi and Mn-SOD in gills and GST-pi in digestive glands had good correlations with B[a]P concentrations and could be used as molecular biomarkers of B[a]P exposure. This study investigated the molecular response of the genes mentioned above and selected useful molecular biomarkers for B[a]P pollution monitoring.

[1]  J. W. Flesher,et al.  Mechanism of aralkyl-DNA adduct formation from benzo[a]pyrene in vivo. , 1994, Chemical research in toxicology.

[2]  T. Roepke,et al.  Activation of multidrug efflux transporter activity at fertilization in sea urchin embryos (Strongylocentrotus purpuratus). , 2004, Developmental biology.

[3]  B. Ketterer,et al.  Theta, a new class of glutathione transferases purified from rat and man. , 1991, The Biochemical journal.

[4]  L. Pan,et al.  Identification of a novel P450 gene belonging to the CYP4 family in the clam Ruditapes philippinarum, and analysis of basal- and benzo(a)pyrene-induced mRNA expression levels in selected tissues. , 2011, Environmental toxicology and pharmacology.

[5]  P. K. Lam,et al.  Antioxidant responses to benzo[a]pyrene and Aroclor 1254 exposure in the green-lipped mussel, Perna viridis. , 2004, Environmental pollution.

[6]  L. Pan,et al.  Cloning, characterization and tissue distribution of a pi-class glutathione S-transferase from clam (Venerupis philippinarum): Response to benzo[alpha]pyrene exposure. , 2010, Comparative biochemistry and physiology. Toxicology & pharmacology : CBP.

[7]  F. Sörgel,et al.  Inhibition of uridinediphosphate glucuronyltransferase caused by furosemide , 1980, Experientia.

[8]  John W. Farrington,et al.  The Mussel Watch , 1978, Environmental Conservation.

[9]  M. Wigler,et al.  Cloning the differences between two complex genomes , 1993, Science.

[10]  L. Pan,et al.  AHH activity, tissue dose and DNA damage in different tissues of the scallop Chlamys farreri exposed to benzo[a]pyrene. , 2008, Environmental pollution.

[11]  David Brown.  Linking Molecular and Population Processes in Mathematical Models of Quorum Sensing , 2013, Bulletin of mathematical biology.

[12]  L. Pan,et al.  Biomarkers and bioaccumulation of clam Ruditapes philippinarum in response to combined cadmium and benzo[α]pyrene exposure. , 2011, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[13]  L. Pan,et al.  Effects of benzo(k)fluoranthene exposure on the biomarkers of scallop Chlamys farreri. , 2005, Comparative biochemistry and physiology. Toxicology & pharmacology : CBP.

[14]  Yun-Fai Chris Lau,et al.  [20] Suppression subtractive hybridization: A versatile method for identifying differentially expressed genes , 1999 .

[15]  D. Mackay,et al.  Controlling persistent organic pollutants-what next? , 1998, Environmental toxicology and pharmacology.

[16]  J. Hayes,et al.  The glutathione S-transferase supergene family: regulation of GST and the contribution of the isoenzymes to cancer chemoprotection and drug resistance. , 1995, Critical reviews in biochemistry and molecular biology.

[17]  F. Oesch,et al.  A fluorometric assay for quantitating phenol sulfotransferase activities in homogenates of cells and tissues. , 1987, Analytical biochemistry.

[18]  C. Elferink,et al.  Role of the aryl hydrocarbon receptor in cell cycle regulation. , 2002, Chemico-biological interactions.

[19]  H. Jörnvall,et al.  Identification of three classes of cytosolic glutathione transferase common to several mammalian species: correlation between structural data and enzymatic properties. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[20]  A. Pardee,et al.  Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. , 1992, Science.

[21]  G. Theilacker,et al.  Transfer of the chlorinated hydrocarbon PCB in a laboratory marine food chain , 1977 .

[22]  L. Pan,et al.  Molecular cloning and sequence analysis and the response of a aryl hydrocarbon receptor homologue gene in the clam Ruditapes philippinarum exposed to benzo(a)pyrene. , 2010, Comparative biochemistry and physiology. Toxicology & pharmacology : CBP.

[23]  W. Jarman,et al.  Characterization of multixenobiotic/multidrug transport in the gills of the mussel Mytilus californianus and identification of environmental substrates , 1995 .

[24]  Yang Yong-liang.  Distribution and sources of polycyclic aromatic hydrocarbon in sediments of Jiaozhou Bay , 2003 .

[25]  G. Gores,et al.  A fluorometric assay for quantitating DNA strand breaks during apoptosis. , 1995, Analytical biochemistry.

[26]  Hae-Chul Park,et al.  A manganese superoxide dismutase (MnSOD) from Ruditapes philippinarum: comparative structural- and expressional-analysis with copper/zinc superoxide dismutase (Cu/ZnSOD) and biochemical analysis of its antioxidant activities. , 2012, Fish & shellfish immunology.

[27]  P. Hebert,et al.  Biological and physical factors affecting the body burden of organic contaminants in freshwater mussels , 1990, Archives of environmental contamination and toxicology.

[28]  P. Degan,et al.  Tissue dose, DNA adducts, oxidative DNA damage and CYP1A-immunopositive proteins in mussels exposed to waterborne benzo[a]pyrene. , 1998, Mutation research.

[29]  Some Thoughts Concerning Quotients, Risks, and Decision-Making , 1996 .

[30]  Structure, function and evolution of glutathione transferases: implications for classification of non-mammalian members of an ancient enzyme superfamily. , 2001 .

[31]  Jianmin Zhao,et al.  Identification and expression profile of a new cytochrome P450 isoform (CYP414A1) in the hepatopancreas of Venerupis (Ruditapes) philippinarum exposed to benzo[a]pyrene, cadmium and copper. , 2012, Environmental toxicology and pharmacology.

[32]  S. Lukyanov,et al.  Suppression subtractive hybridization: a versatile method for identifying differentially expressed genes. , 1999, Methods in enzymology.

[33]  P. Lam,et al.  Relationships between tissue concentrations of polycyclic aromatic hydrocarbons and antioxidative responses of marine mussels, Perna viridis. , 2001, Aquatic toxicology.

[34]  M. Depledge,et al.  Assessment of trace metal toxicity using molecular, physiological and behavioural biomarkers , 1995 .

[35]  Xu Xue-ren.  A preliminary study on PAHs in the surface sediment samples from Dalian Bay , 2001 .

[36]  A. Kong,et al.  Induction of phase I, II and III drug metabolism/transport by xenobiotics , 2005, Archives of pharmacal research.

[37]  D. Nelson,et al.  P450 gene nomenclature based on evolution. , 1991, Methods in enzymology.

[38]  Chen She-jun.  Source, Transport and Risk Assessment of PAHs in Surface Sediments from Pearl River Delta , 2006 .

[39]  Jianmin Zhao,et al.  Expression profiles of seven glutathione S-transferase (GST) genes from Venerupis philippinarum exposed to heavy metals and benzo[a]pyrene. , 2012, Comparative biochemistry and physiology. Toxicology & pharmacology : CBP.

[40]  M. Gilek,et al.  Tissue differences, dose-response relationship and persistence of DNA adducts in blue mussels (Mytilus edulis L) exposed to benzo[a]pyrene. , 2003, Aquatic toxicology.

[41]  L. Pan,et al.  Application of SSH and quantitative real time PCR to construction of gene expression profiles from scallop Chlamys farreri in response to exposure to tetrabromobisphenol A. , 2012, Environmental toxicology and pharmacology.

[42]  E. Cavalieri,et al.  Central role of radical cations in metabolic activation of polycyclic aromatic hydrocarbons. , 1995, Xenobiotica; the fate of foreign compounds in biological systems.

[43]  Taisen Iguchi,et al.  Linking molecular and population stress responses in Daphnia magna exposed to cadmium. , 2008, Environmental science & technology.

[44]  D. Sheehan,et al.  Structure, function and evolution of glutathione transferases: implications for classification of non-mammalian members of an ancient enzyme superfamily. , 2001, The Biochemical journal.

[45]  M. Pfaffl,et al.  A new mathematical model for relative quantification in real-time RT-PCR. , 2001, Nucleic acids research.

[46]  M. Bachmann,et al.  Expression of P-glycoprotein gene in marine sponges. Identification and characterization of the 125 kDa drug-binding glycoprotein. , 1992, Carcinogenesis.