Archives of Toxicology

, Volume 84, Issue 1, pp 3–16 | Cite as

Metabolism of arsenic in human liver: the role of membrane transporters

  • Zuzana Drobná
  • Felecia S. Walton
  • David S. Paul
  • Weibing Xing
  • David J. Thomas
  • Miroslav Stýblo
Review Article


Metabolism of inorganic arsenic (iAs) is one of the key factors determining the character of adverse effects associated with exposure to iAs. Results of previous studies indicate that liver plays a primary role in iAs metabolism. This paper reviews these results and presents new data that link the capacity of human hepatocytes to metabolize iAs to the expression of specific membrane transporters. Here, we examined relationship between the expression of potential arsenic transporters (AQP9, GLUT2, P-gp, MRP1, MRP2, and MRP3) and the production and cellular retention of iAs and its methylated metabolites in primary cultures of human hepatocytes exposed for 24 h to subtoxic concentrations of arsenite. Our results show that the retention of iAs and methylarsenic metabolites (MAs) by hepatocytes exposed to sub-micromolar concentrations of arsenite correlates negatively with MRP2 expression. A positive correlation was found between MRP2 expression and the production of dimethylarsenic metabolites (DMAs), specifically, the concentration of DMAs in culture media. After exposures to high micromolar concentrations of arsenite which almost completely inhibited MAs and DMAs production, a positive correlation was found between the expression of GLUT2 and cellular retention of iAs and MAs. MRP3, AQP9, or P-gp expression had no effect on the production or distribution of iAs, MAs, or DMAs, regardless of the exposure level. Hepatocytes from seven donors used in this study did not contain detectable amounts of MRP1 protein. These data suggest that MRP2 plays an important role in the efflux of DMAs, thus, regulating kinetics of the methylation reactions and accumulation of iAs and MAs by human hepatocytes. The membrane transport of iAs by high-capacity GLUT2 transporters is not a rate-limiting step for the metabolism of arsenite at low exposure level, but may play a key role in accumulation of iAs after acute exposures which inhibit iAs methylation.


Arsenic Human liver Membrane transport 


  1. Benbrahim-Tallaa L, Waterland RA, Styblo M, Achanzar WE, Webber MM, Waalkes MP (2005) Molecular events associated with arsenic induced malignant transformation of human prostatic epithelial cells: aberrant genomic DNA methylation and oncogene activation. Toxicol Appl Pharmacol 206:288–298CrossRefPubMedGoogle Scholar
  2. Benramdane L, Accominotti M, Fanton L, Malicier D, Vallon J-J (1999) Arsenic speciation in human organs following fatal arsenic trioxide poisoning—a case report. Clin Chem 45:301–306PubMedGoogle Scholar
  3. Bhattacharjee H, Carbrey J, Rosen BP, Mukhopadhyay R (2004) Drug uptake and pharmacological modulation of drug sensitivity in leukemia by AQP9. Biochem Biophys Res Commun 322:836–841CrossRefPubMedGoogle Scholar
  4. Brambila EM, Achanzar WE, Qu W, Webber MM, Waalkes MP (2002) Chronic arsenic-exposed human prostate epithelial cells exhibit stable arsenic tolerance: mechanistic implications of altered cellular glutathione and glutathione S-transferase. Toxicol Appl Pharmacol 183:99–107CrossRefPubMedGoogle Scholar
  5. Bredfeldt TG, Jagadish B, Eblin KE, Mash EA, Gandolfi AJ (2006) Monomethylarsonous acid induces transformation of human bladder cells. Toxicol Appl Pharmacol 216:69–79CrossRefPubMedGoogle Scholar
  6. Buchet JP, Lauwerys R (1985) Study of inorganic arsenic methylation by rat in vitro: relevance for the interpretation of observations in man. Arch Toxicol 57:125–129CrossRefPubMedGoogle Scholar
  7. Buchet JP, Geubel A, Pauwels S, Mahieu P, Lauwerys R (1984) The influence of liver disease on the methylation of arsenite in humans. Arch Toxicol 55:151–154CrossRefPubMedGoogle Scholar
  8. Cohen SM, Ohnishi T, Arnold LL, Le XC (2007) Arsenic-induced bladder cancer in an animal model. Toxicol Appl Pharmacol 222:258–263CrossRefPubMedGoogle Scholar
  9. Cullen WR, McBride BC, Reglinski J (1984) The reaction of methylarsenicals with thiols: some biological implications. J Inorg Biochem 21:179–194CrossRefGoogle Scholar
  10. Delnomdedieu M, Basti MM, Otvos JD, Thomas DJ (1994) Reduction and binding of arsenate and dimethylarsenate by glutathione: a multinuclear magnetic resonance study. Chem-Biol Interact 90:139–155CrossRefPubMedGoogle Scholar
  11. Devesa V, Del Razo LM, Adair B, Drobná Z, Waters SB, Hughes MF, Styblo M, Thomas DJ (2004) Comprehensive analysis of arsenic metabolites by pH-specific hydride generation atomic absorption spectrometry. J Anal At Spectrom 19:1460–1467CrossRefGoogle Scholar
  12. Drobná Z, Jaspers I, Thomas DJ, Styblo M (2003) Differential activation of AP-1 in human bladder epithelial cells by inorganic and methylated arsenicals. FASEB J 17:67–69PubMedGoogle Scholar
  13. Drobná Z, Waters SB, Walton FS, LeCluyse EL, Thomas DJ, Styblo M (2004) Interindividual variation in the metabolism of arsenic in cultured primary human hepatocytes. Toxicol Appl Pharmacol 201:166–177CrossRefPubMedGoogle Scholar
  14. Drobná Z, Waters SB, Devesa V, Harmon AW, Thomas DJ, Stýblo M (2005) Metabolism and toxicity of As in human urothelial cells expressing rat arsenic (+3 oxidation state) methyltransferase. Toxicol Appl Pharmacol 207:147–159CrossRefPubMedGoogle Scholar
  15. Drobná Z, Xing W, Thomas DJ, Stýblo M (2006) shRNA silencing of AS3MT expression minimizes arsenic methylation capacity of HepG2 cells. Chem Res Toxicol 19:894–898CrossRefPubMedGoogle Scholar
  16. Geubel AP, Mairlot MC, Buchet JP, Dive C, Lauwerys R (1988) Abnormal methylation capacity in human liver cirrhosis. Int J Pharm Res VIII(2):117–122Google Scholar
  17. Healy SM, Wildfang E, Zakharyan RA, Aposhian HV (1999) Diversity of inorganic arsenite biotransformation. Biol Trace Element Res 68:249–266CrossRefGoogle Scholar
  18. Hepner GW, Vesell ES (1975) Quantitative assessment of hepatic function by breath analysis after oral administration of (14C)aminopyrine. Ann Internal Med 83:632–638Google Scholar
  19. Hernández A, Xamena N, Sekaran C, Tokunaga H, Sampayo-Reyes A, Quinteros D, Creus A, Marcos R (2008a) High arsenic metabolic efficiency in AS3MT287Thr allele carriers. Pharmacogenet Genomics 18:349–355CrossRefPubMedGoogle Scholar
  20. Hernández A, Xamena N, Surrallés J, Sekaran C, Tokunaga H, Quinteros D, Creus A, Marcos R (2008b) Role of the Met(287)Thr polymorphism in the AS3MT gene on the metabolic arsenic profile. Mutat Res 637:80–92PubMedGoogle Scholar
  21. Hirata M, Mohri T, Hisanaga A, Ishinishi N (1989) Conversion of arsenite and arsenate to methylarsenic and dimethylarsenic compounds by homogenates prepared from livers and kidneys of rats and mice. Appl Organomet Chem 3:335–341CrossRefGoogle Scholar
  22. Hoffmaster KA, Turncliff RZ, LeCluyse EL, Kim RB, Meier PJ, Brouwer KLR (2004) P-glycoprotein expression, localization, and function in sandwich-cultured primary rat and human hepatocytes: relevance to the hepatobiliary disposition of a model opioid peptide. Pharm Res 21:1294–1302CrossRefPubMedGoogle Scholar
  23. Kala SV, Neely MW, Kala G, Prater CI, Atwood DW, Rice JS, Lieberman MW (2000) The MRP2/cMOAT transporter and arsenic-glutathione complex formation are required for biliary excretion of arsenic. J Biol Chem 275:33404–33408CrossRefPubMedGoogle Scholar
  24. Kiermayer C, Michalke B, Schmidt J, Brielmeier M (2007) Effect of selenium on thioredoxin reductase activity in Txnrd1 or Txnrd2 hemizygous mice. Biol Chem 388:1091–1097CrossRefPubMedGoogle Scholar
  25. Konig J, Rost D, Cui Y, Keppler D (1999) Characterization of the human multidrug resistance protein isoform MRP3 localized to the basolateral hepatocyte membrane. Hepatology 29:1156–1163CrossRefPubMedGoogle Scholar
  26. Lee S, Kim J, Kwon K, Yoon HW, Levine RL, Ginsburg A, Rhee SG (1999) Molecular cloning and characterization of a mitochondrial selenocysteine-containing thioredoxin reductase from rat liver. J Biol Chem 274:4722–4734CrossRefPubMedGoogle Scholar
  27. Lee TC, Ho IC, Lu WJ, Huang JD (2006) Enhanced expression of Multidrug resistance-associated protein 2 and reduced expression of Aquaglyceroporin 3 in am arsenic-resistant human cell line. J Biol Chem 281:18401–18407CrossRefPubMedGoogle Scholar
  28. Leung J, Pang A, Yuen WH, Kwong YL, Tse EWC (2007) Relationship of expression of aquaglyceroporin 9 with arsenic uptake and sensitivity in leukemia cells. Blood 109:740–746CrossRefPubMedGoogle Scholar
  29. Lin S, Cullen WR, Thomas DJ (1999) Methylarsenicals and arsinothiols are potent inhibitors of mouse liver thioredoxin reductase. Chem Res Toxicol 12:924–930CrossRefPubMedGoogle Scholar
  30. Lin S, Shi Q, Nix FB, Styblo M, Beck MA, Herbin-Davis KM, Hall LL, Simeonsson JB, Thomas DJ (2002) A novel S-Adenosyl-L-methionine: arsenic(III) methyltransferase from rat liver cytosol. J Biol Chem 277:10795–10803CrossRefPubMedGoogle Scholar
  31. Lindberg AL, Kumar R, Goessler W, Thirumaran R, Gurzau E, Koppova K, Rudnai P, Leonardi G, Fletcher T, Vahter M (2007) Metabolism of low-dose inorganic arsenic in a central European population: influence of sex and genetic polymorphisms. Environ Health Perspect 115:1081–1086PubMedGoogle Scholar
  32. Liu J, Chen H, Miller DS, Saavedra JE, Keefer LK, Johnson DR, Klaassen CD, Waalkes MP (2001) Overexpression of glutathione S-transferase II and multidrug resistance transport proteins is associated with acquired tolerance to inorganic arsenic. Mol Pharmacol 60:302–309PubMedGoogle Scholar
  33. Liu Z, Shen J, Carbrey JM, Mukhopadhyay R, Agre P, Rosen BP (2002) Arsenite transport by mammalian aquaglyceroporins AQP7 and AQP9. Proc Natl Acad Sci USA 99:6053–6058CrossRefPubMedGoogle Scholar
  34. Liu Z, Carbrey JM, Agre P, Rosen BP (2004) Arsenic trioxide uptake by human and rat aquaglyceroporins. Biochem Biophys Res Commun 316:1178–1185CrossRefPubMedGoogle Scholar
  35. Liu Z, Styblo M, Rosen BP (2006a) Methylarsonous acid transport by aquaglyceroporins. Environ Health Perspect 114:527–531PubMedCrossRefGoogle Scholar
  36. Liu Z, Sanchez MA, Jiang X, Boles E, Landfear SM, Rosen BP (2006b) Mammalian glucose permease GLUT1 facilitates transport of arsenic trioxide and methylarsonous acid. Biochem Biophys Res Commun 351:424–430CrossRefPubMedGoogle Scholar
  37. Lu WJ, Tamai I, Nezu J, Lai ML, Huang JD (2006) Organic anion transporting polypeptide-C mediates arsenic uptake in HEK-293 cells. J Biomed Sci 13:525–533CrossRefPubMedGoogle Scholar
  38. Mahagita C, Grassl SM, Piyachaturawat P, Ballatori N (2007) Human organic anion transporter 1B1 and 1B3 function as bidirectional carriers and do not mediate GSH-bile acid cotransport. Am J Physiol Gastrointest Liver Physiol 293:G271–G278CrossRefPubMedGoogle Scholar
  39. Maheu P, Buchet JP, Roels HA, Lauwerys R (1987) The metabolism of arsenic in humans acutely intoxicated by As2O3. Clin Toxicol 18:1067–1075CrossRefGoogle Scholar
  40. Marafante E, Vahter M, Envall J (1985) The role of methylation in the detoxification of arsenate in the rabbit. Chem-Biol Interact 56:225–238CrossRefPubMedGoogle Scholar
  41. Mass MJ, Tennant A, Roop BC, Cullen WR, Styblo M, Thomas DJ, Kligerman AD (2001) Methylated trivalent arsenic species are genotoxic. Chem Res Toxicol 14:355–361CrossRefPubMedGoogle Scholar
  42. Moriwaki Y, Yamamoto T, Higashino K (1999) Enzymes involved in purine metabolism—a review of histochemical localization and functional implications. Histol Histopathol 14:1321–1340PubMedGoogle Scholar
  43. Mure K, Uddin AN, Lopez LC, Styblo M, Rossman TG (2003) Arsenite induces delayed mutagenesis and transformation in human osteosarcoma cells at extremely low concentrations. Environ Mol Mutagen 41:322–331CrossRefPubMedGoogle Scholar
  44. Naranmandura H, Ibata K, Suzuki KT (2007) Toxicity of dimethylmonothioarsinic acid toward human epidermoid carcinoma A431 cells. Chem Res Toxicol 2:1120–1125CrossRefGoogle Scholar
  45. Németi B, Csanaky I, Gregus Z (2006) Effect of an inactivator of glyceraldehyde-3-phosphate dehydrogenase, a fortuitous arsenate reductase, on disposition of arsenate in rats. Tox Sci 90:49–60CrossRefGoogle Scholar
  46. Paul DS, Harmon AW, Devesa V, Thomas DJ, Styblo M (2007) Molecular mechanisms of diabetogenic effects of arsenic: inhibition of insulin signaling by arsenite and methylarsonous acid. Environ Health Perspect 115:734–742PubMedGoogle Scholar
  47. Payen L, Courtois A, Campion JP, Guillouzo A, Fardel O (2000) Characterization and inhibition by a wide range of xenobiotics of organic anion excretion by primary human hepatocytes. Biochem Pharmacol 60:1967–1975CrossRefPubMedGoogle Scholar
  48. Petrick JS, Ayala-Fierro F, Cullen WR, Carter DE, Aposhian VH (2000) Monomethylarsonous acid (MMA(III)) is more toxic than arsenite in Chang human hepatocytes. Toxicol Appl Pharmacol 163:203–207CrossRefPubMedGoogle Scholar
  49. Petrick JS, Jagadish B, Mash EA, Aposhian HV (2001) Monomethylarsonous acid (MMAIII) and arsenite: LD50 in hamsters and in vitro inhibition of pyruvate dehydrogenase. Chem Res Toxicol 14:651–656CrossRefPubMedGoogle Scholar
  50. Planchamp C, Hadengue A, Stieger B, Bourquin J, Vonlaufen A, Frossard JL, Quadri R, Becker CD, Pastor CM (2007) Function of both sinusoidal and canalicular transporters controls the concentration of organic anions within hepatocytes. Mol Pharmacol 71:1089–1097CrossRefPubMedGoogle Scholar
  51. Raml R, Rumpler A, Goessler W, Vahter M, Li L, Ochi T, Francesconi KA (2007) Thio-dimethylarsinate is a common metabolite in urine samples from arsenic-exposed women in Bangladesh. Toxicol Appl Pharmacol 222:374–380CrossRefPubMedGoogle Scholar
  52. Reay PF, Asher CJ (1977) Preparation and purification of 74As-labeled arsenate and arsenite for use in biological experiments. Anal Biochem 78:557–560CrossRefPubMedGoogle Scholar
  53. Roach PJ (2002) Glycogen and its metabolism. Curr Mol Med 2:101–120CrossRefPubMedGoogle Scholar
  54. Sens DA, Park S, Gurel V, Sens MA, Garrett SH, Somji S (2004) Inorganic cadmium- and arsenite-induced malignant transformation of human bladder urothelial cells. Toxicol Sci 79:56–63CrossRefPubMedGoogle Scholar
  55. Styblo M, Thomas DJ (1997) Binding of arsenicals to proteins in an in vitro methylation system. Toxicol Appl Pharmacol 147:1–8CrossRefPubMedGoogle Scholar
  56. Styblo M, Delnomdedieu M, Thomas DJ (1995a) Biological mechanism and toxicological consequences of the methylation of arsenic. In: Goyer RA, Cherian MG (eds) Handbook of experimental pharmacology, vol 115, toxicology of metals—biochemical aspects. Springer, New York, pp 407–433Google Scholar
  57. Styblo M, Yamauchi H, Thomas DJ (1995b) Comparative in vitro methylation of trivalent and pentavalent arsenic species. Toxicol Appl Pharmacol 135:172–178CrossRefPubMedGoogle Scholar
  58. Styblo M, Serves SV, Cullen WR, Thomas DJ (1997) Comparative inhibition of yeast glutathione reductase by arsenicals and arsenothiols. Chem Res Toxicol 10:27–33CrossRefPubMedGoogle Scholar
  59. Styblo M, Del Razo LM, LeCluyse EL, Hamilton GA, Wang C, Cullen WR, Thomas DJ (1999) Metabolism of arsenic in primary cultures of human and rat hepatocytes. Chem Res Toxicol 12:560–565CrossRefPubMedGoogle Scholar
  60. Styblo M, Del Razo LM, Vega L, Germolec DR, LeCluyse EL, Hamilton GA, Reed W, Wang C, Cullen WR, Thomas DJ (2000) Comparative toxicity of trivalent and pentavalent inorganic and methylated arsenicals in human cells. Arch Toxicol 74:289–299CrossRefPubMedGoogle Scholar
  61. Styblo M, Drobná Z, Jaspers I, Lin S, Thomas DJ (2002) The role of biomethylation in toxicity and carcinogenicity of arsenic. A research update. Environ Health Perspect 110(Suppl 5):767–771PubMedGoogle Scholar
  62. Takeshita A, Shinjo K, Naito K, Matsui H, Shigeno K, Nakamura S, Horii T, Maekawa M, Kitamura K, Naoe T, Ohmishi K, Ohno R (2003) P-glycoprotein (P-gp) and multidrug resistance-associated protein 1 (MRP1) are induced by arsenic trioxide (As2O3), but are not the main mechanism of As2O3-resistance in acute promyelotic leukemia cells. Leukemia 17:648–650CrossRefPubMedGoogle Scholar
  63. Thomas DJ, Styblo M, Shan L (2001) Cellular metabolism and systemic toxicity of arsenic. Toxicol Appl Pharmacol 176:127–144CrossRefPubMedGoogle Scholar
  64. Thomas DJ, Li J, Waters SB, Xing W, Adair BM, Drobna Z, Devesa V, Styblo M (2007) Arsenic (+3 oxidation state) methyltransferase and methylation of arsenicals. Exp Biol Med 232:3–13Google Scholar
  65. Vega L, Styblo M, Patterson R, Cullen W, Wang C, Germolec D (2001) Differential effects of trivalent and pentavalent arsenicals on cell proliferation and cytokine secretion in normal human epidermal keratinocytes. Toxicol Appl Pharmacol 172:225–232CrossRefPubMedGoogle Scholar
  66. Vernhet L, Seite MP, Allain N, Guillouzo A, Fardel O (2001) Arsenic induces expression of the multidrug resistance-associated protein 2 (MRP2) gene in primary rat and human hepatocytes. J Pharmacol Exp Ther 298:234–239PubMedGoogle Scholar
  67. Walton FS, Waters SB, Jolley SL, LeCluyse EL, Thomas DJ, Styblo M (2003) Selenium compounds modulate the activity of recombinant rat AsIII-methyltransferase and the methylation of arsenite by rat and human hepatocytes. Chem Res Toxicol 16:261–265CrossRefPubMedGoogle Scholar
  68. Walton FS, Harmo AW, Paul DS, Drobná Z, Patel YM, Styblo M (2004) Inhibition of insulin-dependent glucose uptake by trivalent arsenicals: possible mechanism of arsenic-induced diabetes. Toxicol Appl Pharmacol 198:424–433CrossRefPubMedGoogle Scholar
  69. Waters SB, Devesa-Perez V, Del Razo LM, Styblo M, Thomas DJ (2004a) Endogenous reductants support catalytic function of recombinant rat cyt19, an arsenic methyltransferase. Chem Res Toxicol 17:404–409CrossRefPubMedGoogle Scholar
  70. Waters SB, Devesa V, Fricke MW, Creed JT, Styblo M, Thomas DJ (2004b) Glutathione modulates recombinant rat arsenic (+3 oxidation state) methyltransferase-catalyzed formation of trimethylarsine oxide and trimethylarsine. Chem Res Toxicol 17:1621–1629CrossRefPubMedGoogle Scholar
  71. Wood TC, Salavagionne O, Mukherjee B, Wang L, Klumpp AF, Thomae BA, Eckloff BW, Schaid DJ, Wieben EC, Weinshilboum RM (2006) Human arsenic methyltransferase (AS3MT) pharmacogenetics: gene resequencing and functional genomics studies. J Biol Chem 281:7364–7373CrossRefPubMedGoogle Scholar
  72. Xie Y, Liu J, Liu Y, Klaassen CD, Waalkes MP (2004) Toxicokinetic and genomic analysis of chronic arsenic exposure in multidrug-resistance mdr1a/1b(−/−) double knockout mice. Mol Cell Biochem 255:11–18CrossRefPubMedGoogle Scholar
  73. Yin ZL, Dahlstrom JE, Le Couteur DG, Board PG (2001) Immunohistochemistry of omega class glutathione S-transferase in human tissues. J Histochem Cytochem 49:983–987PubMedGoogle Scholar
  74. Zhou L, Jing Y, Styblo M, Chen Z, Waxman S (2005) Glutathione S-transferase π inhibits As2O3-induced apoptosis in lymphoma cells: involvement of hydrogen peroxide. Blood 105:1198–1203CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Zuzana Drobná
    • 1
  • Felecia S. Walton
    • 1
  • David S. Paul
    • 1
  • Weibing Xing
    • 2
    • 3
  • David J. Thomas
    • 4
  • Miroslav Stýblo
    • 1
    • 5
  1. 1.Department of NutritionUniversity of North Carolina at Chapel HillChapel HillUSA
  2. 2.Curriculum in ToxicologyThe University of North Carolina at Chapel HillChapel HillUSA
  3. 3.Duke Clinical Research InstituteDuke University Medical Center, Duke UniversityDurhamUSA
  4. 4.Integrated Systems Toxicology Division, National Health and Environmental Effects Research Laboratory, Office of Research and DevelopmentUS Environmental Protection AgencyResearch Triangle ParkUSA
  5. 5.Center for Environmental Medicine, Asthma, and Lung BiologyThe University of North Carolina at Chapel HillChapel HillUSA

Personalised recommendations