Overexpression of CYP3A5 attenuates inducibility and activity of CYP3A4 in HepG2 cells

Kuang,Zemin; Huang,Zhijun; Li,Ying; Yang,Guoping; Liu,Meilin; Yuan,Hong

doi:10.3892/mmr.2014.3022

Overexpression of CYP3A5 attenuates inducibility and activity of CYP3A4 in HepG2 cells

Authors:
- Zemin Kuang
- Zhijun Huang
- Ying Li
- Guoping Yang
- Meilin Liu
- Hong Yuan
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Affiliations: Department of Hypertension, Beijing Anzhen Hospital of Capital Medical University, Beijing 100029, P.R. China, Center of Clinical Pharmacology, The Third Xiangya Hospital, Central South University, Changsha, Hunan 410013, P.R. China, Department of Geriatrics, The First Hospital, Peking University, Beijing 100034, P.R. China
Published online on: December 1, 2014 https://doi.org/10.3892/mmr.2014.3022
Pages: 2868-2874

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Abstract

There have been conflicting reports regarding the catalytic role of cytochrome P450 (CYP)3A5, which range from deeming it irrelevant to suggesting it is equally as important as CYP3A4, the most potent and abundant catalytic cytochrome enzyme in the human liver. This was partially attributed to the fact that CYP3A5 is highly polymorphic. However the importance of other underlying mechanisms remain unclear. The aim of the present study was to investigate the interaction between these enzymes. A human HepG2 hepatocellular line stably overexpressing CYP3A5 was constructed. The results suggested that CYP3A5 does not affect CYP3A4 expression directly. However, overexpression of CYP3A5 attenuated the inducibility of CYP3A4 in response to dexamethasone. A luciferase reporter assay indicated that this attenuation was due to a decrease in CYP3A4 promoter activity. Furthermore, a pharmacokinetic assay using quinidine and amlodipine showed that CYP3A4 enzyme activity per mg of microsomal protein was also decreased in the group overexpressing CYP3A5 compared with the dexamethasone‑induced control group. In conclusion, the current study demonstrated that CYP3A5 may affect CYP3A4 at the transcriptional level and may thus modify CYP3A4 expression and activity in the presence of substrates and inducers. The results indicate that CYPs may interact with each other under certain conditions and that this interaction may be a novel mechanism by which drug‑drug interactions are mediated.

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1	Kruijtzer CM, Beijnen JH and Schellens JH: Improvement of oral drug treatment by temporary inhibition of drug transporters and/or cytochrome P450 in the gastrointestinal tract and liver: an overview. Oncologist. 7:516–530. 2002. View Article : Google Scholar : PubMed/NCBI
2	Casarett LJ and Doull J: Toxicology: The Basic Science of Poisons. Macmillan Publishing Company; London, UK: 1975
3	Anzenbacher P and Anzenbacherová E: Cytochromes P450 and metabolism of xenobiotics. Cell Mol Life Sci. 58:737–747. 2001. View Article : Google Scholar : PubMed/NCBI
4	Stevens JC, Hines RN, Gu C, et al: Developmental expression of the major human hepatic CYP3A enzymes. J Pharmacol Exp Ther. 307:573–582. 2003. View Article : Google Scholar : PubMed/NCBI
5	Luo G, Cunningham M, Kim S, et al: CYP3A4 induction by drugs: correlation between a pregnane X receptor reporter gene assay and CYP3A4 expression in human hepatocytes. Drug Metab Dispos. 30:795–804. 2002. View Article : Google Scholar : PubMed/NCBI
6	Katoh M, Nakajima M, Yamazaki H and Yokoi T: Inhibitory effects of CYP3A4 substrates and their metabolites on P-glycoprotein-mediated transport. Eur J Pharm Sci. 12:505–513. 2001. View Article : Google Scholar : PubMed/NCBI
7	Kivistö KT, Niemi M and Fromm MF: Functional interaction of intestinal CYP3A4 and P-glycoprotein. Fundam Clin Pharmacol. 18:621–626. 2004. View Article : Google Scholar : PubMed/NCBI
8	Galetin A, Burt H, Gibbons L and Houston JB: Prediction of time-dependent CYP3A4 drug-drug interactions: impact of enzyme degradation, parallel elimination pathways, and intestinal inhibition. Drug Metab Dispos. 34:166–175. 2006. View Article : Google Scholar
9	Wang RW, Newton DJ, Liu N, Atkins WM and Lu AY: Human cytochrome P-450 3A4: in vitro drug-drug interaction patterns are substrate-dependent. Drug Metab Dispos. 28:360–366. 2000.PubMed/NCBI
10	Walsky RL, Obach RS, Hyland R, et al: Selective mechanism-based inactivation of CYP3A4 by CYP3cide (PF-04981517) and its utility as an in vitro tool for delineating the relative roles of CYP3A4 versus CYP3A5 in the metabolism of drugs. Drug Metab Dispos. 40:1686–1697. 2012. View Article : Google Scholar : PubMed/NCBI
11	Yuan R, Madani S, Wei XX, Reynolds K and Huang SM: Evaluation of cytochrome P450 probe substrates commonly used by the pharmaceutical industry to study in vitro drug interactions. Drug Metab Dispos. 30:1311–1319. 2002. View Article : Google Scholar : PubMed/NCBI
12	Huang W, Lin YS, McConn DJ II, et al: Evidence of significant contribution from CYP3A5 to hepatic drug metabolism. Drug Metab Dispos. 32:1434–1445. 2004. View Article : Google Scholar : PubMed/NCBI
13	Dai Y, Hebert MF, Isoherranen N, et al: Effect of CYP3A5 polymorphism on tacrolimus metabolic clearance in vitro. Drug Metab Dispos. 34:836–847. 2006. View Article : Google Scholar : PubMed/NCBI
14	Kivistö KT, Niemi M, Schaeffeler E, et al: CYP3A5 genotype is associated with diagnosis of hypertension in elderly patients: data from DEBATE study. Am J Pharmacogenomics. 5:191–195. 2005. View Article : Google Scholar
15	Dai Y, Iwanaga K, Lin YS, et al: In vitro metabolism of cyclosporine A by human kidney CYP3A5. Biochem Pharmacol. 68:1889–1902. 2004. View Article : Google Scholar : PubMed/NCBI
16	Kuehl P, Zhang J, Lin Y, et al: Sequence diversity in CYP3A promoters and characterization of the genetic basis of polymorphic CYP3A5 expression. Nat Genet. 27:383–391. 2001. View Article : Google Scholar : PubMed/NCBI
17	Zhang YP, Zuo XC, Huang ZJ, et al: CYP3A5 polymorphism, amlodipine and hypertension. J Hum Hypertens. 28:145–149. 2013. View Article : Google Scholar : PubMed/NCBI
18	Shi Y, Li Y, Tang J, et al: Influence of CYP3A4, CYP3A5 and MDR-1 polymorphisms on tacrolimus pharmacokinetics and early renal dysfunction in liver transplant recipients. Gene. 512:226–231. 2013. View Article : Google Scholar
19	Zuo XC, Ng CM, Barrett JS, et al: Effects of CYP3A4 and CYP3A5 polymorphisms on tacrolimus pharmacokinetics in Chinese adult renal transplant recipients: a population pharmacokinetic analysis. Pharmacogenet Genomics. 23:251–261. 2013. View Article : Google Scholar : PubMed/NCBI
20	Suzuki Y, Itoh H, Fujioka T, et al: Association of plasma concentration of 4β-hydroxycholesterol with CYP3A5 polymorphism and plasma concentration of indoxyl sulfate in stable kidney transplant recipients. Drug Metab Dispos. 42:105–110. 2014. View Article : Google Scholar
21	Bhatnagar V, Garcia EP, O’Connor DT, et al: CYP3A4 and CYP3A5 polymorphisms and blood pressure response to amlodipine among African-American men and women with early hypertensive renal disease. Am J Nephrol. 31:95–103. 2010. View Article : Google Scholar :
22	Kim KA, Park PW, Lee OJ, et al: Effect of CYP3A5^*3 genotype on the pharmacokinetics and pharmacodynamics of amlodipine in healthy Korean subjects. Clin Pharmacol Ther. 80:646–656. 2006. View Article : Google Scholar : PubMed/NCBI
23	Overbergh L, Valckx D, Waer M and Mathieu C: Quantification of murine cytokine mRNAs using real time quantitative reverse transcriptase PCR. Cytokine. 11:305–312. 1999. View Article : Google Scholar : PubMed/NCBI
24	Paine MF, Khalighi M, Fisher JM, et al: Characterization of interintestinal and intraintestinal variations in human CYP3A-dependent metabolism. J Pharmacol Exp Ther. 283:1552–1562. 1997.
25	Galetin A, Brown C, Hallifax D, Ito K and Houston JB: Utility of recombinant enzyme kinetics in prediction of human clearance: impact of variability, CYP3A5, and CYP2C19 on CYP3A4 probe substrates. Drug Metab Dispos. 32:1411–1420. 2004. View Article : Google Scholar : PubMed/NCBI
26	Ghosh SS, Basu AK, Ghosh S, et al: Renal and hepatic family 3A cytochromes P450 (CYP3A) in spontaneously hypertensive rats. Biochem Pharmacol. 50:49–54. 1995. View Article : Google Scholar : PubMed/NCBI
27	Zhang L, Miyaki K, Wang W and Muramatsu M: CYP3A5 polymorphism and sensitivity of blood pressure to dietary salt in Japanese men. J Hum Hypertens. 24:345–350. 2010. View Article : Google Scholar
28	Ho H, Pinto A, Hall SD, et al: Association between the CYP3A5 genotype and blood pressure. Hypertension. 45:294–298. 2005. View Article : Google Scholar
29	Riffel AK, Schuenemann E and Vyhlidal CA: Regulation of the CYP3A4 and CYP3A7 promoters by members of the nuclear factor I transcription factor family. Mol Pharmacol. 76:1104–1114. 2009. View Article : Google Scholar : PubMed/NCBI
30	Maruyama M, Matsunaga T, Harada E and Ohmori S: Comparison of basal gene expression and induction of CYP3As in HepG2 and human fetal liver cells. Biol Pharm Bull. 30:2091–2097. 2007. View Article : Google Scholar : PubMed/NCBI
31	Sarkar MA and Jackson BJ: Theophylline N-demethylations as probes for P4501A1 and P4501A2. Drug Metab Dispos. 22:827–834. 1994.PubMed/NCBI
32	Badyal DK and Dadhich AP: Cytochrome P450 and drug interactions. Ind J Pharmacol. 33:248–259. 2001.
33	Moore LB, Parks DJ, Jones SA, et al: Orphan nuclear receptors constitutive androstane receptor and pregnane X receptor share xenobiotic and steroid ligands. J Biol Chem. 275:15122–15127. 2000. View Article : Google Scholar : PubMed/NCBI
34	Drocourt L, Ourlin JC, Pascussi JM, Maurel P and Vilarem MJ: Expression of CYP3A4, CYP2B6, and CYP2C9 is regulated by the vitamin D receptor pathway in primary human hepatocytes. J Biol Chem. 277:25125–25132. 2002. View Article : Google Scholar : PubMed/NCBI
35	Mao Z, Luan X, Cao G, et al: DEC1 binding to the proximal promoter of CYP3A4 ascribes to the downregulation of CYP3A4 expression by IL-6 in primary human hepatocytes. Biochem Pharmacol. 84:701–711. 2012. View Article : Google Scholar : PubMed/NCBI
36	Tirona RG, Lee W, Leake BF, et al: The orphan nuclear receptor HNF4alpha determines PXR-and CAR-mediated xenobiotic induction of CYP3A4. Nat Med. 9:220–224. 2003. View Article : Google Scholar : PubMed/NCBI
37	Matsumura K, Saito T, Takahashi Y, et al: Identification of a novel polymorphic enhancer of the human CYP3A4 gene. Mol Pharmacol. 65:326–334. 2004. View Article : Google Scholar : PubMed/NCBI
38	Jover R, Bort R, Gómez-Lechón MJ and Castell JV: Down-regulation of human CYP3A4 by the inflammatory signal interleukin-6: molecular mechanism and transcription factors involved. FASEB J. 16:1799–1801. 2002.PubMed/NCBI
39	Kacevska M, Ivanov M, Wyss A, et al: DNA methylation dynamics in the hepatic CYP3A4 gene promoter. Biochimie. 94:2338–2344. 2012. View Article : Google Scholar : PubMed/NCBI
40	Busi F and Cresteil T: CYP3A5 mRNA degradation by nonsense mediated mRNA decay. Mol Pharmacol. 68:808–815. 2005.PubMed/NCBI
41	Hu YF, He J, Chen GL, et al: CYP3A5^3 and CYP3A4^18 single nucleotide polymorphisms in a Chinese population. Clin Chim Acta. 353:187–192. 2005. View Article : Google Scholar : PubMed/NCBI
42	Chou FC, Tzeng SJ and Huang JD: Genetic polymorphism of cytochrome P450 3A5 in Chinese. Drug Metab Dispos. 29:1205–1209. 2001.PubMed/NCBI
43	Jounaïdi Y, Guzelian PS, Maurel P and Vilarem MJ: Sequence of the 5′-flanking region of CYP3A5: comparative analysis with CYP3A4 and CYP3A7. Biochem Biophys Res Commun. 205:1741–1747. 1994. View Article : Google Scholar
44	Zeigler-Johnson C, Friebel T, Walker AH, et al: CYP3A4, CYP3A5, and CYP3A43 genotypes and haplotypes in the etiology and severity of prostate cancer. Cancer Res. 64:8461–8467. 2004. View Article : Google Scholar : PubMed/NCBI
45	Bajpai P, Tripathi AK and Agrawal D: Genetic polymorphism of CYP3A5 in Indian chronic myeloid leukemia patients. Mol Cell Biochem. 336:49–54. 2010. View Article : Google Scholar
46	Krusekopf S, Roots I and Kleeberg U: Differential drug-induced mRNA expression of human CYP3A4 compared to CYP3A5, CYP3A7 and CYP3A43. Eur J Pharmacol. 466:7–12. 2003. View Article : Google Scholar : PubMed/NCBI
47	Lu C and Li AP: Species comparison in P450 induction: effects of dexamethasone, omeprazole, and rifampin on P450 isoforms 1A and 3A in primary cultured hepatocytes from man, Sprague-Dawley rat, minipig, and beagle dog. Chem Biol Interact. 134:271–281. 2001. View Article : Google Scholar : PubMed/NCBI
48	Pascussi JM, Drocourt L, Gerbal-Chaloin S, et al: Dual effect of dexamethasone on CYP3A4 gene expression in human hepatocytes. Sequential role of glucocorticoid receptor and pregnane X receptor. Eur J Biochem. 268:6346–6358. 2001. View Article : Google Scholar : PubMed/NCBI
49	Roberts PJ, Rollins KD, Kashuba AD, et al: The influence of CYP3A5 genotype on dexamethasone induction of CYP3A activity in African Americans. Drug Metab Dispos. 36:1465–1469. 2008. View Article : Google Scholar : PubMed/NCBI
50	Pirmohamed M, James S, Meakin S, et al: Adverse drug reactions as cause of admission to hospital: prospective analysis of 18,820 patients. BMJ. 329:15–19. 2004. View Article : Google Scholar : PubMed/NCBI
51	Zhao Y, Song M, Guan D, et al: Genetic polymorphisms of CYP3A5 genes and concentration of the cyclosporine and tacrolimus. Transplant Proc. 37:178–181. 2005. View Article : Google Scholar : PubMed/NCBI