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The Pharmaceutical Society of Korea (대한약학회)
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Histone Deacetylase Inhibitor Stimulate CYP3A4 Proximal Promoter Activity in HepG2 Cells

  • Published : 2004.04.01
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Abstract

The expression of CYP3A4 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 of CYP3A4 gene expression have not been understood. In order to gain the insight of the molecular mechanism of CYP3A4 gene expression, study has been undertaken to investigate if the histone deacetylation is involved in the regulation of CYP3A4 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 plasmid PCYP3A4-Luc containing ${\~}1kb$ of the CYP3A4 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 of CYP3A4 proximal promoter activity in the absence of SXR and histone deacetylase (HDAC) inhibitors. 4-Dimethylamino-H-[4-(2-hydroxycarbamoylvinyl)benzyl]benzamide (IN2001), a new class HDAC inhibitor significantly increased CYP3A4 proximal promoter activity over untreated control cells and rifampicin concomitant treatment with IN2001 increased further CYP3A4 proximal promoter activity that was stimulated by IN2001 The results of this study demon-strated that both HDAC inhibitors and SXR are essential to increase of CYP3A4 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 stimulate CYP3A4 proximal promoter activity. Also this data suggested that HDAC inhibitors seemed to facilitate the CYP3A4 proximal promoter to be activated by chemicals.

Keywords

References

  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 human CYP3A4 and rabbit CYP3A6 in primary cultures of adult rat and rabbitHepa-Itocytes. Mol. Pharmacal., 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-Itocytes. J. Biol. Chem., 277,25125-25132 (2002) https://doi.org/10.1074/jbc.M201323200
  3. Gibson, G. G., Regulation of the CYP3A4 gene by hydrocortisone and xenobiotics: Role of the glucocorticoid and pregnane X receptors. Drug Metab. Dispos., 28,493-496 (2000)
  4. Goodwin, S., Hodgson, E., and Liddle, C., The orphan human pregnane X receptor mediates the transcriptional activation of CYP3A4 by rifampicin through a distal enhancer module. Mol. Pharmacal., 56,1329-1339 (1999) https://doi.org/10.1124/mol.56.6.1329
  5. Guengerich, F. P., Cytochrome p-450 3A4: Regulation and role in drug metabolism, Annu. Rev. Pharmacol. Toxicol., 39,1-17 (1999) https://doi.org/10.1146/annurev.pharmtox.39.1.1
  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 P450in human livers, and its transcriptional control. Eur. J. Biochem., 218, 585-595 (1993) https://doi.org/10.1111/j.1432-1033.1993.tb18412.x
  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-4501l1A7 (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). https://doi.org/10.1016/0167-4781(92)90520-A
  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 psychopharmacology. J. Clinic. Psychopharmacol., 15, 387-398 (1995) https://doi.org/10.1097/00004714-199512000-00002
  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 HepaItocytes. Drug Metab. Dispos., 23, 415-421 (1995)
  10. Li, A. P., Kaminski, D. L., and Rasmussen, A., Substrates of human Hepa-Itic cytochrome P450 3A4. Toxicology, 104, 1-8 (1995) https://doi.org/10.1016/0300-483X(95)03155-9
  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-Itocytes. J. Clin. Endocrinol. Metab., 83, 2411-2416 (1998) https://doi.org/10.1210/jc.83.7.2411
  12. Luo, G., Cunningham, M., Kim S., Burn, 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-Itocytes. Drug Metab. Dispos., 30, 795-804 (2002) https://doi.org/10.1124/dmd.30.7.795
  13. Mangelsdorf, D. J., and Evans, R. M., The RXR heterodimers and orphan receptors. Cell, 83, 841-850 (1995) https://doi.org/10.1016/0092-8674(95)90200-7
  14. Marks, P. A., Miller, T., and Richon, V. M., Histone deacetylases. Curr. Opin. Pharmacol., 3, 344-351 (2003) https://doi.org/10.1016/S1471-4892(03)00084-5
  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) https://doi.org/10.1128/MCB.21.7.2298-2311.2001
  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) https://doi.org/10.1016/S0378-4274(03)00216-9
  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 human CYP3A4 gene in vitro. Xenobiotica, 29, 269-279 (1999) https://doi.org/10.1080/004982599238669
  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) https://doi.org/10.1016/S0304-4165(02)00483-X
  20. Pazin, M. J., and Kadonaga, J. T., Whats up and down with histone deacetylation and transcription? Cell, 89, 325-328 (1997) https://doi.org/10.1016/S0092-8674(00)80211-1
  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) https://doi.org/10.1002/hep.1840180535
  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) https://doi.org/10.1074/jbc.274.10.6043
  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) https://doi.org/10.1038/87912
  24. Takeshita, A, Taguchi, M., Koibuchi, N., and Ozawa, Y., Putative role of the orphan nuclear receptor SXR in the mechanism of CYP3A4 inhibition by Xenobiotics. J. Biol. Chem., 277, 32453-32458 (2002) https://doi.org/10.1074/jbc.M111245200
  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) https://doi.org/10.1124/mol.60.6.1399
  26. Wu, C., Chromatin remodeling and the control of gene expression. J. Biol. Chem., 272, 28171-28174 (1997) https://doi.org/10.1074/jbc.272.45.28171