Genotyping Study of Cytochrome P450 2A6 Alel CYP2A6*1 and CYP2A6*9 among Javanese Indonesian Smokers
Abstract
CYP2A6 belongs to the CYP2 family of P450 cytochromes were highly polimorphy. CYP2A6*1 (wild type) was an active allele, CYP2A6*9 was agen decreased allele and the CYP2A6*4 was an inactive allele. By using the polymerase chain reaction technique (PCR), the CYP2A6 polymorphism was studied among Javanese Indonesian smokers. In the process of genotyping the CYP2A6*9 allele, some subjects who had been genotyped as CYP2A6*1/*4 in our previous studies were regenotyped as CYP2A6*9 in this study. The primer forward 2A6*9S and the primer reverse 2A6*9AS-wild type were used in these study. The Promega Go Taq Green Master Mix reagent were used to amplifl y the allele CYP2A6*1 in the positions at -395 to -28 of the CYP2A6 gene. The sample studied consisted of 20 smokers with Cigarrete per-Day (CPD) <10 and 13 smoker with CPD 11-20 from Javanese Indonesian population. In theses research, the subjects had been genotyped as CYP2A6*1/*4 in our previous studies. The allele frequencies of CYP2A6*1, CYP2A6*4, and CYP2A6*9 were 48.5%, 48.5%, and 3%, respectively. When these allele were considered simultaneously, among the subject, 63.9% were genotyped for CYP2A6*1/*4 and 6.1% were genotyped for CYP2A6*1/*4/*9. Based on the data collected, it could be concluded that the polymorphism of CYP2A6 among Javanese population sample study was not aff ected on smoking behavior.
References
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27. Gambier N, Batt a-M, Marie B, Pfi ster M, Siest G, Visvikis-Siest S. Association of CYP2A6*1B genetic variant with the amount of smoking in French adults from the Stanislas cohort. Pharmacogenomics J. 2005.5(4):271–5.
28. Martini S. Makna merokok pada remaja putri perokok. 2014.3(2):119–27. 29. Wismanto, B. Sarwo B. Strategi Penghentian Perilaku Merokok. Semarang: Unika Soegijapranata. 2007;21– 2.
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31. Chenoweth MJ, O’Loughlin J, Sylvestre M-P, Tyndale RF. CYP2A6 slow nicotine metabolism is associated with increased quitting by adolescent smokers. Pharmacogenet Genomics. 2013.23(4):232–5.
32. Ikeda K, Yoshisue K, Matsushima E, Nagayama S, Kobayashi K, Tyson CA, et al. Bioactivation of tegafur to 5-fl uorouracil is catalyzed by cytochrome P-450 2A6 in human liver microsomes in vitro. Clin Cancer Res. 2000.6(11):4409–15.
33. Daigo S, Takahashi Y, Fujieda M, Ariyoshi N, Yamazaki H, Koizumi W, et al. A novel mutant allele of the CYP2A6 gene (CYP2A6*11 ) found in a cancer patient who showed poor metabolic phenotype towardstegafur. Pharmacogenetics. 2002.12(4):299–306.
34. Djordjevic N, Carrillo JA, Gervasini G, Jankovic S, Aklillu E. In vivo evaluation of CYP2A6 and xanthine oxidase enzyme activities in the Serbian population. Eur J Clin Pharmacol. 2010.66(6):571–8.
35. Kimura M, Yamazaki H, Fujieda M, Kiyotani K, Honda G, Saruwatari J, et al. Cyp2a6 is a principal enzyme involved in hydroxylation of 1,7-dimethylxanthine, a main caff eine metabolite, in humans. Drug Metab Dispos. 2005.33(9):1361–6.
36. Arab-Alameddine M, Di Iulio J, Buclin T, Rotger M, Lubomirov R, Cavassini M, et al. Pharmacogeneticsbased population pharmacokinetic analysis of efavirenz in HIV-1-infected individuals. Clin Pharmacol Ther. 2009.85(5):485–94.
37. Iulio J di, Fayet A, Arab-Alameddine M, Rotger M, Lubomirov R, Cavassini M, et al. In vivo analysis of efavirenz metabolism in individuals with impaired CYP2A6 function. Pharmacogenet Genomics. 2009.19(4):300–9.
38. Tan L, Yu J-T, Sun Y-P, Ou J-R, Song J-H, Yu Y. The influence of cytochrome oxidase CYP2A6, CYP2B6, and CYP2C9 polymorphisms on the plasma concentrations of valproic acid in epileptic patients. Clin Neurol Neurosurg. 2010.112(4):320–3.
39. Endo T, Nakajima M, Fukami T, Hara Y, Hasunuma T, Yokoi T, et al. Genetic polymorphisms of CYP2A6 aff ect the in-vivo pharmacokinetics of pilocarpine. Pharmacogenet Genomics. 2008.18(9):761–72.
40. Endo T, Ban M, Hirata K, Yamamoto A, Hara Y, Momose Y. Involvement of CYP2A6 in the formation of a novel metabolite, 3-hydroxypilocarpine, from pilocarpine in human liver microsomes. Drug Metab Dispos. 2007.35(3):476–83.
41. Nunoya KI, Yokoi T, Kimura K, Kainuma T, Satoh K, Kinoshita M, et al. A new CYP2A6 gene deletion responsible for the in vivo polymorphic metabolism of (+)-cis-3,5-dimethyl-2-(3-pyridyl)thiazolidin-4- one hydrochloride in humans. J Pharmacol Exp Ther. 1999.289(1):437–42.
42. Desta Z, Kreutz Y, Nguyen AT, Li L, Skaar T, Kamdem LK, et al. Plasma letrozole concentrations in postmenopausal women with breast cancer are associated with CYP2A6 genetic variants, body mass index, and age. Clin Pharmacol Ther. 2011.90(5):693– 700.
2. Karaconji IB. Facts about nicotine toxicity. Arh Hig Rada Toksikol. 2005.56(4):363–71.
3. Oscarson M. Genetic polymorph isms in th e cytochrome P450 2A6 (CYP2A6) gene: implications for interindividual diff erences in nicotine metabolism. Drug Metab Dispos. 2001.29(2):91–5.
4. Raunio H, Rautio A, Gullstén H, Pelkonen O. Polymorphisms of CYP2A6 and its practical consequences. Br J Clin Pharmacol. 2001.52(4):357– 63.
5. Yoshida R, Nakajima M, Watanabe Y, Kwon J-T, Yokoi T. Genetic polymorphisms in human CYP2A6gene causing impaired nicotine metabolism. Br J Clin Pharmacol. 2002.54(5):511–7.
6. Hukkanen J, Jacob P 3rd, Benowitz NL. Metabolism and disposition kinetics of nicotine. Pharmacol Rev. 2005.57(1):79–115.
7. Kwon JT, Nakajima M, Chai S, Yom YK, Kim HK, Yamazaki H, et al. Nicotine metabolism and CYP2A6 allele frequencies in Koreans. Pharmacogenetics. 2001.
8. Nakajima M, Kwon JT, Tanaka N, Zenta T, Yamamoto Y, Yamamoto H, et al. Relationship between interindividual diff erences in nicotine metabolism and CYP2A6 genetic polymorphism in humans. Clin Pharmacol Ther. 2001.69(1):72–8.
9. Oscarson M, McLellan RA, Gullstén H, Yue QY, Lang MA, Bernal ML, et al. Characterisation and PCRbased detection of a CYP2A6 gene deletion found at a high frequency in a Chinese population. FEBS Lett. 1999.448(1):105–10.
10. Rao Y, Hoff mann E, Zia M, Bodin L, Zeman M, Sellers EM, et al. Duplications and defects in the CYP2A6 gene: identifi cation, genotyping, and in vivo eff ects on smoking. Mol Pharmacol. 2000.58(4):747–55. 11. Yusof W, Gan SH. High prevalence of CYP2A6*4 and CYP2A6*9 alleles detected among a Malaysian population. Clin Chim Acta. 2009.403(1–2):105–9.
12. Nakajima M, Yokoi T. Interindividual variability in nicotine metabolism: C-oxidation and glucuronidation. Drug Metab Pharmacokinet. 2005.20(4):227–35.
13. Ando M, Hamajima N, Ariyoshi N, Kamataki T, Matsuo K, Ohno Y. Association of CYP2A6 gene deletion with cigarette smoking status in Japanese adults. J Epidemiol. 2003.13(3):176–81.
14. Ariyoshi N, Miyamoto M, Umetsu Y, Kunitoh H, Dosaka-Akita H, Sawamura Y-I, et al. Genetic polymorphism of CYP2A6 gene and tobacco-induced lung cancer risk in male smokers. Cancer Epidemiol biomarkers Prev a Publ Am Assoc Cancer Res cosponsored by Am Soc Prev Oncol. 2002.11(9):890–4.
15. Fujieda M, Yamazaki H, Saito T, Kiyotani K, Gyamfi MA, Sakurai M, et al. Evaluation of CYP2A6 genetic polymorphisms as determinants of smoking behavior and tobacco-related lung cancer risk in male Japanese smokers. Carcinogenesis. 2004.25(12):2451–8.
16. Minematsu N, Nakamura H, Furuuchi M, Nakajima T, Takahashi S, Tateno H, et al. Limitation of cigarette consumption by CYP2A6*4, *7 and *9 polymorphisms. Eur Respir J. 2006.27(2):289–92.
17. Kadlubar S, Anderson JP, Sweeney C, Gross MD, Lang NP, Kadlubar FF, et al. Phenotypic CYP2A6 variation and the risk of pancreatic cancer. JOP. 2009.10(3):263–70.
18. Liu Y, Xu Y, Li F, Chen H, Guo S. CYP2A6 deletion polymorphism is associated with decreased susceptibility of lung cancer in Asian smokers: a metaanalysis. Tumour Biol J Int Soc Oncodevelopmental Biol Med. 2013.34(5):2651–7.
19. Minematsu N, Nakamura H, Iwata M, Tateno H, Nakajima T, Takahashi S, et al. Association of CYP2A6 deletion polymorphism with smoking habitand development of pulmonary emphysema. Thorax. 2003.58(7):623–8.
20. Wang L, Zang W, Liu J, Xie D, Ji W, Pan Y, et al. Association of CYP2A6*4 with susceptibility of lung cancer: a meta-analysis. PLoS One. 2013.8(4):e59556.
21. Patramurti C, Nurrochmad A, Martono S, Science P, Mada G, Chemistry P. Polymorphism of cytochrome P450 2A6 (CYP2A6*1 and CYP2A6*4 ) among Javanese Indonesian Smoker. 2015.26(1):11–9.
22. Yoshida, R., Nakajima, M., Nishimura, K., Tokudome, S., Kwon, J.-T., dan Yokoi T. Eff ects of polymorphism in promoter region of human CYP2A6 gene ( CYP2A6 * 9 ) on expression level of messenger ribonucleic acid and enzymatic activity in vivo and in vitro. 2003.9236(3):69–76.
23. Mwenifumbo JC, Sellers EM, Tyndale RF. Nicotine metabolism and CYP2A6 activity in a population of black African descent: Impact of gender and light smoking. Drug Alcohol Depend. 2007.89(1):24–33.
24. Schoedel K a, Hoff mann EB, Rao Y, Sellers EM, Tyndale RF. Ethnic variation in CYP2A6 and association of genetically slow nicotine metabolism and smoking in adult Caucasians. Pharmacogenetics. 2004.14(9):615–26.
25. Ingelman-Sundberg M, Oscarson M, Daly a K, Garte S, Nebert DW. Human cytochrome P-450 (CYP) genes: a web page for the nomenclature of alleles. Cancer Epidemiol Biomarkers Prev. 2001.10(12):1307–8.
26. Liu T, Chen W-Q, David SP, Tyndale RF, Wang H, Chen Y-M, et al. Interaction between heavy smoking and CYP2A6 genotypes on type 2 diabetes and its possible pathways. Eur J Endocrinol. 2011.165(6):961–7.
27. Gambier N, Batt a-M, Marie B, Pfi ster M, Siest G, Visvikis-Siest S. Association of CYP2A6*1B genetic variant with the amount of smoking in French adults from the Stanislas cohort. Pharmacogenomics J. 2005.5(4):271–5.
28. Martini S. Makna merokok pada remaja putri perokok. 2014.3(2):119–27. 29. Wismanto, B. Sarwo B. Strategi Penghentian Perilaku Merokok. Semarang: Unika Soegijapranata. 2007;21– 2.
30. Ray R, Tyndale RF, Lerman C. Nicotine dependence pharmacogenetics: role of genetic variation in nicotinemetabolizing enzymes. J Neurogenet. 2009.23(3):252– 61.
31. Chenoweth MJ, O’Loughlin J, Sylvestre M-P, Tyndale RF. CYP2A6 slow nicotine metabolism is associated with increased quitting by adolescent smokers. Pharmacogenet Genomics. 2013.23(4):232–5.
32. Ikeda K, Yoshisue K, Matsushima E, Nagayama S, Kobayashi K, Tyson CA, et al. Bioactivation of tegafur to 5-fl uorouracil is catalyzed by cytochrome P-450 2A6 in human liver microsomes in vitro. Clin Cancer Res. 2000.6(11):4409–15.
33. Daigo S, Takahashi Y, Fujieda M, Ariyoshi N, Yamazaki H, Koizumi W, et al. A novel mutant allele of the CYP2A6 gene (CYP2A6*11 ) found in a cancer patient who showed poor metabolic phenotype towardstegafur. Pharmacogenetics. 2002.12(4):299–306.
34. Djordjevic N, Carrillo JA, Gervasini G, Jankovic S, Aklillu E. In vivo evaluation of CYP2A6 and xanthine oxidase enzyme activities in the Serbian population. Eur J Clin Pharmacol. 2010.66(6):571–8.
35. Kimura M, Yamazaki H, Fujieda M, Kiyotani K, Honda G, Saruwatari J, et al. Cyp2a6 is a principal enzyme involved in hydroxylation of 1,7-dimethylxanthine, a main caff eine metabolite, in humans. Drug Metab Dispos. 2005.33(9):1361–6.
36. Arab-Alameddine M, Di Iulio J, Buclin T, Rotger M, Lubomirov R, Cavassini M, et al. Pharmacogeneticsbased population pharmacokinetic analysis of efavirenz in HIV-1-infected individuals. Clin Pharmacol Ther. 2009.85(5):485–94.
37. Iulio J di, Fayet A, Arab-Alameddine M, Rotger M, Lubomirov R, Cavassini M, et al. In vivo analysis of efavirenz metabolism in individuals with impaired CYP2A6 function. Pharmacogenet Genomics. 2009.19(4):300–9.
38. Tan L, Yu J-T, Sun Y-P, Ou J-R, Song J-H, Yu Y. The influence of cytochrome oxidase CYP2A6, CYP2B6, and CYP2C9 polymorphisms on the plasma concentrations of valproic acid in epileptic patients. Clin Neurol Neurosurg. 2010.112(4):320–3.
39. Endo T, Nakajima M, Fukami T, Hara Y, Hasunuma T, Yokoi T, et al. Genetic polymorphisms of CYP2A6 aff ect the in-vivo pharmacokinetics of pilocarpine. Pharmacogenet Genomics. 2008.18(9):761–72.
40. Endo T, Ban M, Hirata K, Yamamoto A, Hara Y, Momose Y. Involvement of CYP2A6 in the formation of a novel metabolite, 3-hydroxypilocarpine, from pilocarpine in human liver microsomes. Drug Metab Dispos. 2007.35(3):476–83.
41. Nunoya KI, Yokoi T, Kimura K, Kainuma T, Satoh K, Kinoshita M, et al. A new CYP2A6 gene deletion responsible for the in vivo polymorphic metabolism of (+)-cis-3,5-dimethyl-2-(3-pyridyl)thiazolidin-4- one hydrochloride in humans. J Pharmacol Exp Ther. 1999.289(1):437–42.
42. Desta Z, Kreutz Y, Nguyen AT, Li L, Skaar T, Kamdem LK, et al. Plasma letrozole concentrations in postmenopausal women with breast cancer are associated with CYP2A6 genetic variants, body mass index, and age. Clin Pharmacol Ther. 2011.90(5):693– 700.
Published
2017-04-30
How to Cite
PATRAMURTI, CHRISTINE; FENTY, FENTY.
Genotyping Study of Cytochrome P450 2A6 Alel CYP2A6*1 and CYP2A6*9 among Javanese Indonesian Smokers.
JURNAL ILMU KEFARMASIAN INDONESIA, [S.l.], v. 15, n. 1, p. 50-56, apr. 2017.
ISSN 2614-6495.
Available at: <http://jifi.farmasi.univpancasila.ac.id/index.php/jifi/article/view/453>. Date accessed: 30 aug. 2025.
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