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Histone deacetylase inhibitor stimulateCYP3A4 proximal promoter activity in hepg2 cells


The expression ofCYP3A4 gene is induced by a variety of structurally unrelated xenobiotics including the antibiotic rifampicin, pregnenolone 16-carbonitrile (PCN), and endogenous hormones, that might mediate through steroid and xenobiotic receptor (SXR) system. The molecular mechanisms underlying regulation ofCYP3A4 gene expression have not been understood. In order to gain the insight of the molecular mechanism ofCYP3A4 gene expression, study has been undertaken to investigate if the histone deacetylation is involved in the regulation ofCYP3A4 gene expression by proximal promoter in human hepatoma HepG2 cells. Also we have investigated to see if SXR is involved in the regulation of CYP3A4 proximal promoter activity in human hepatoma HepG2 cells. HepG2 cells were transfected with a plasmidpCYP3A4-Luc containing~1 kb of theCYP3A4 proximal promoter region (-863 to +64 bp) in front of a reporter gene, luciferase, in the presence or absence of pSAP-SXR. In HepG2 cells, CYP3A4 inducers, such as rifampicin, PCN and RU486 showed minimal stimulation ofCYP3A4 proximal promoter activity in the absence of SXR and histone deacetylase (HDAC) inhibitors. 4-Dimethylamino-N-[4-(2-hydroxycarbamoylvinyl)benzyl]benzamide (IN2001), a new class HDAC inhibitor significantly increasedCYP3A4 proximal promoter activity over untreated control cells and rifampicin concomitant treatment with IN2001 increased furtherCYP3A4 proximal promoter activity that was stimulated by IN2001. The results of this study demonstrated that both HDAC inhibitors and SXR are essential to increase ofCYP3A4 proximal promoter activity by CYP3A4 inducers such as PCN, rifampicin, and RU486. Especially SXR seems to be important for the dose dependent response of CYP3A4 inducing chemicals to stimulateCYP3A4 proximal promoter activity. Also this data suggested that HDAC inhibitors seemed to facilitate theCYP3A4 proximal promoter to be activated by chemicals.

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  1. Barwick, J. L., Quattrochi, L. C., Mills, A. S., Potenza, C., Tukey, R. H., and Guzelian, P. S., Trans-species gene transfer for analysis of glucocorticoid-inducible transcriptional activation of transiently expressed humanCYP3A4 and rabbitCYP3A6 in primary cultures of adult rat and rabbit Hepa-ltocytes.Mol. Pharmacol., 50, 10–16 (1996).

  2. Drocourt, L., Ourlin, J. C., Pascussi, J. M., Maurel, P., and Vilarem, M. J., Expression of CYP3A4, CYP2B6, and CYP2C9 is regulated by the vitamin D receptor pathway in primary human Hepa-ltocytes.J. Biol. Chem., 277, 25125–25132 (2002).

  3. Gibson, G. G., Regulation of theCYP3A4 gene by hydrocortisone and xenobiotics: Role of the glucocorticoid and pregnane X receptors.Drug Metab. Dispos., 28, 493–496 (2000).

  4. Goodwin, B., Hodgson, E., and Liddle, C., The orphan human pregnane X receptor mediates the transcriptional activation ofCYP3A4 by rifampicin through a distal enhancer module.Mol. Pharmacol., 56, 1329–1339 (1999).

  5. Guengerich, F. P., Cytochrome p-450 3A4: Regulation and role in drug metabolism,Annu. Rev. Pharmacol. Toxicol., 39, 1–17 (1999).

  6. Hashimoto, H., Toide, K., Kitamura, R., Fujita, M., Tagawa, S., Itoh, S., and Kamataki, T., Gene structure of CYP3A4, an adult-specific form of cytochrome P450 in human livers, and its transcriptional control.Eur. J. Biochem., 218, 585–595 (1993).

  7. Itoh, S., Yanagimoto, T., Tagawa, S., Hashimoto, H., Kitamura, R., Nakajima, Y., Okochi, T., Fujimoto, S., Uchino, J., and Kamataki, T., Genomic organization of human fetal specific P-450IIIA7 (cytochrome P-450HFLa)-related gene(s) and interaction of transcriptional regulatory factor with its DNA element in the 5′ flanking region.Biochim. Biophys. Acta, 1130, 133–138 (1992).

  8. Ketter, T. A., Flockhart, D. A., Post, R. M., Denicoff, K., Pazzaglia, P. J., Marangell, L. B., George, M. S., and Callahan, A. M., The emerging role of cytochrome P450 3A in psycho-pharmacology.J. Clinic. Psychopharmacol., 15, 387–398 (1995).

  9. Kocarek, T. A., Schuetz, E. G., Strom, S. C., Fisher, R. A., and Guzelian, P. S., Comparative analysis of cytochrome P4503A induction in primary cultures of rat, rabbit, and human Hepa-Itocytes.Drug Metab. Dispos., 23, 415–421 (1995).

  10. Li, A. P., Kaminski, D. L., and Rasmussen, A., Substrates of human Hepa-ltic cytochrome P450 3A4.Toxicology, 104, 1–8 (1995).

  11. Liddle, C., Goodwin, B. J., George, J., Tapner, M., and Farrell, G. C., Separate and interactive regulation of cytochrome P450 3A4 by triiodothyronine, dexamethasone, and growth hormone in cultured Hepa-ltocytes.J. Clin. Endocrinol. Metab., 83, 2411–2416 (1998).

  12. Luo, G., Cunningham, M., Kim S., Bum, T., Lin, J., Sinz, M., Hamilton, G., Rizzo, C., Jolley, S., Gilbert, D., Downey, A., Mudra, D., Graham, R., Carroll, K., Xie, J., Madan, A., Parkinson, A., Christ, D., Selling, B., Lecluyse, E., and Gan, L., CYP3A4 induction by drugs: Correlation between a pregnane X receptor reporter gene assay and CYP3A4 expression in human Hepa-ltocytes.Drug Metab. Dispos., 30, 795–804 (2002).

  13. Mangelsdorf, D. J., and Evans, R. M., The RXR heterodimers and orphan receptors.Cell, 83, 841–850 (1995).

  14. Marks, P. A., Miller, T., and Richon, V. M., Histone deacetylases.Curr. Opin. Pharmacol., 3, 344–351 (2003).

  15. Mathur, M., Tucker P. W., and Samuels, H. H., PSF is a novel corepressor that mediates its effect through Sin3A and the DNA binding domain of nuclear hormone receptors.Mol. Cell Biol., 21, 2298–2311 (2001).

  16. Michalets, E. L., Update: clinically significant cytochrome P-450 drug interactions.Pharmacotherapy, 18, 84–112 (1998).

  17. Nakajima, A., Iwanari, M., and Yokoi, T., Effects of histone deacetylation and DNA methylation on the constitutive and TCDD-inducible expressions of the human CYP1 family in MCF-7 and HeLa cells.Toxicol. Lett., 144, 247–256 (2003).

  18. Ogg, M. S., Williams, J. M., Tarbit, M., Goldfarb, P. S., Grays, T. J. B., and Gibson, G. G., A reporter gene assay to assess the molecular mechanisms of xenobiotic-dependent induction of the humanCYP3A4 genein vitro.Xenobiotica, 29, 269–279 (1999).

  19. Pascussi, J. M., Gerbal-Chaloin, S., Drocourt, L., Maurel, P., and Vilarem, M. J., The expression of CYP2B6, CYP2C9 and CYP3A4 genes, a tangle of networks of nuclear and steroid receptors.Biochim. Biophysic. Act., 1619, 243–253 (2003).

  20. Pazin, M. J., and Kadonaga, J. T., Whats up and down with histone deacetylation and transcription?Cell, 89, 325–328 (1997).

  21. Schuetz, E. G., Schuetz, J. D., Strom, S. C., Thompson, M. T., Fisher, R. A., Molowa, D. T., Li, D., and Guzelian, P. S., Regulation of human liver cytochromes P-450 in family 3A in primary and continuous culture of human Hepatocytes.Hepatology 18, 1254–1262 (1993).

  22. Sueyoshi, T., Kawamoto, T., Zelko, I., Honkakoski, P., and Negishi, M., The repressed nuclear receptor CAR responds to phenobarbital in activating the human CYP2B6 gene.J. Biol. Chem., 274, 6043–6046 (1999).

  23. Synold, T. W., Dussault, I., and Forman, B. M., The orphan nuclear receptor SXR coordinately regulates drug metabolism and efflux.Nat. Med., 7, 584–590 (2001).

  24. Takeshita, A., Taguchi, M., Koibuchi, N., and Ozawa, Y., Putative role of the orphan nuclear receptor SXR in the mechanism ofCYP3A4 inhibition by Xenobiotics.J. Biol. Chem., 277, 32453–32458 (2002).

  25. Thummel, K. E., Brimer, C., Yasuda, K., Thottassery, J., Senn, T., Lin, Y., Ishizuka, H., Kharasch, E., Schuetz, J., and Schuetz, E., Transcriptional control of intestinal cytochrome P-4503A by 1alpha,25-dihydroxy vitamin D3.Mol. Pharmacol., 60, 1399–1406 (2001).

  26. Wu, C., Chromatin remodeling and the control of gene expression.J. Biol. Chem., 272, 28171–28174 (1997).

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Correspondence to Ja Young Kim or Mee Ryung Ahn or Dae-Kee Kim or Yhun Yhong Sheen.

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Kim, J.Y., Ahn, M.R., Kim, D. et al. Histone deacetylase inhibitor stimulateCYP3A4 proximal promoter activity in hepg2 cells. Arch Pharm Res 27, 407 (2004).

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Key words

  • CYP3A4
  • PCN
  • Rifampicin
  • RU486
  • SXR
  • HDAC
  • IN2001
  • HepG2