Relationship between the 3435C>tTand 2677G>T/A polymorphysms and ABCB1 expression in peripheral blood mononuclear cells extracted from Thai volunteers
DOI:
https://doi.org/10.14456/tjg.2011.2Keywords:
ABCB1 expression, 3435C>T, 2677G>T/A, peripheral blood mononuclear cells (PBMCs)Abstract
Limited data of ABCB1 3435C>T and 2677G>T/A SNPs and its expression in peripheral mononuclear cells (PBMCs) with proper normalization of endogenous controls used is available in Thai population. The primary objective was to investigate the relationship of these SNPs and its expression in PBMCs extracted from Thai healthy individuals. Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) was utilized to determine the SNPs in 30 females and 26 males. Total RNA was extracted from PBMCs followed by reverse transcription. Actin-β, β-2M and GAPDH were internal controls. Real-time quantitative PCR was performed to determine the expression levels. We found that TT genotype of 3435C>T SNP is correlated with higher of ABCB1 expression levels with a significant covariate of gender. 3435C>T genotypes show a strong relationship to 2677G>T genotypes. Most subjects who carry TT genotype at position 3435 will also carry TT genotype at position 2677. No gender difference was noted in the distributions of these SNPs. The results show that genotypes of SNP 3435C>T and gender have, in part, influenced on the expression levels. Therefore, one who carries TT genotype of 3435C>T SNP may have lower response to drugs which target sites are PBMCs.References
Ameyaw, M.M., Regateiro, F., Li, T., Liu, X., Tariq, M., Mobarek, A., Thornton, N., Folayan, G.O., Githang'a, J., Indalo, A., Ofori-Adjei, D., Price-Evans, D.A. and McLeod, H.L. 2001. MDR1 pharmacogenetics: frequency of the C3435T mutation in exon 26 is significantly influenced by ethnicity. Pharmacogenetics 11: 217–221.
Cascorbi, I., Gerloff, T., Johne, A., Meisel, C., Hoffmeyer, S., Schwab, M., Schaeffeler, E., Eichelbaum, M., Brinkmann, U. and Roots, I. 2001. Frequency of single nucleotide polymorphisms in the P-glycoprotein drug transporter MDR1 gene in white subjects. Clin Pharmacol Ther 69: 169–174.
Calado, R.T., Falcão, R.P., Garcia, A.B., Gabellini, S.M., Zago, M.A. and Franco R.F. 2002. Influence of functional MDR1 gene polymorphisms on P-glycoprotein activity in CD34+ hematopoietic stem cells. Haematologica 87: 564–568.
Chowbay, B., Cumaraswamy, S., Cheung, Y.B., Zhou, Q. and Lee E.J. 2003. Genetic polymorphisms in MDR1 and CYP3A4 genes in Asians and the influence of MDR1 haplotypes on cyclosporine disposition in heart transplant recipients. Pharmacogenetics 13: 89–95.
Fellay, J., Marzolini, C., Meaden, E.R., Back, D.J., Buclin, T., Chave, J.P., Decosterd, L.A, Furrer, H., Opravil, M., Pantaleo, G., Retelska, D., Ruiz L., Schinkel, A.H., Vernazza, P., Eap C.B., Telenti, A. 2002. Swiss HIV Cohort Study. Response to antiretroviral treatment in HIV-1-infected individuals with allelic variants of the multidrug resistance transporter 1: a pharmacogenetics study. Lancet 359: 30–36.
Furuno, T., Landi, M.T., Ceroni, M., Caporaso, N., Bernucci, I., Nappi, G., Martignoni, E., Schaeffeler. E., Eichelbaum, M., Schwab, M. and Zanger, U.M. 2002. Expression polymorphism of the blood-brain barrier component P-glycoprotein (MDR1) in relation to Parkinson's disease. Pharmacogenetics 12: 529–534.
Gaikovitch, E.A., Cascorbi, I., Mrozikiewicz, P.M., Brockmöller, J., Frötschl, R., Köpke, K. Gerloff, T., Chernov, J.N. and Roots, I. 2003. Polymorphisms of drug-metabolizing enzymes CYP2C9, CYP2C19, CYP2D6, CYP1A1, NAT2 and of P-glycoprotein in a Russian population. Eur J Clin Pharmacol 59: 303–312.
Goto, M., Masuda, S., Saito, H., Uemoto, S., Kiuchi, T., Tanaka, K. and Inui, K. 2002. C3435T polymorphism in the MDR1 gene affects the enterocyte expression level of CYP3A4 rather than Pgp in recipients of living-donor liver transplantation. Pharmacogenetics 12: 451–457.
Hitzl, M., Drescher, S., van der Kuip, H., Schäffeler, E., Fischer, J., Schwab, M., Eichelbaum, M. and Fromm, M.F. 2001. The C3435T mutation in the human MDR1 gene is associated with altered efflux of the P-glycoprotein substrate rhodamine 123 from CD56+ natural killer cells. Pharmacogenetics 11: 293–298.
Hoffmeyer, S., Burk, O., von Richter, O., Arnold, H.P., Brockmöller, J., Johne A., Cascorbi, I., Gerloff, T., Roots, I., Eichelbaum, M. and Brinkmann, U. 2000. Functional polymorphisms of the human multidrug-resistance gene: multiple sequence variations and correlation of one allele with P-glycoprotein expression and activity in vivo. Proc Natl Acad Sci USA 97: 3473–3478.
Hoffmann, U. and Kroemer, H.K. 2004. The ABC transporters MDR1 and MRP2: multiple functions in disposition of xenobiotics and drug resistance. Drug Metab Rev 36: 669–701.
Horinouchi, M., Sakaeda, T., Nakamura, T., Morita, Y., Tamura, T., Aoyama, N., Kasuga., M. and Okumura, K. 2002. Significant genetic linkage of MDR1 polymorphisms at positions 3435 and 2677: functional relevance to pharmacokinetics of digoxin. Pharm Res 19: 581–585.
Illmer, T., Schuler, U.S., Thiede, C., Schwarz, U.I., Kim, R.B., Gotthard, S., Freund, D., Schäkel, U., Ehninger, G. and Schaich, M. 2002. MDR1 gene polymorphisms affect therapy outcome in acute myeloid leukemia patients. Cancer Res 62: 4955–4962.
Ito, S., Ieiri, I., Tanabe, M., Suzuki, A., Higuchi, S. and Otsubo, K. 2001. Polymorphism of the ABC transporter genes, MDR1, MRP1 and MRP2/cMOAT, in healthy Japanese subjects. Pharmacogenetics 11: 175–184.
Kaya, P., Gündüz, U., Arpaci, F., Ural, A.U. and Guran, S. 2005. Identification of polymorphisms on the MDR1 gene among Turkish population and their effects on multidrug resistance in acute leukemia patients. Am J Hematol 80: 26–34.
Kim, R.B., Leake, B.F., Choo, E.F., Dresser, G.K., Kubba, S.V., Schwarz, U.I., Taylor, A., Xie, H.G., McKinsey, J., Zhou, S., Lan, L.B., Schuetz, J.D., Schuetz, E.G. and Wilkinson, G.R. 2001. Identification of functionally variant MDR1 alleles among European Americans and African Americans. Clin Pharmacol Ther 70: 189–199.
Kimchi-Sarfaty, C., Oh, J.M., Kim, I.W., Sauna, Z.E., Calcagno, A.M., Ambudkar, S. V. and Gottesman, M.M. 2007. A "silent" polymorphism in the MDR1 gene changes substrate specificity. Science 315: 525–528.
Li, Y., Wang, Y., Sun, J., Li, Y. and Yang, L. 2006.
Distribution of the functional MDR1 C3435T polymorphism in the Han population of China. Swiss Med Wkly 136: 377–382.
Moriya, Y., Nakamura, T., Horinouchi, M., Sakaeda, T., Tamura, T., Aoyama, N. Shirakawa, T., Gotoh, A., Fujimoto, S., Matsuo, M., Kasuga, M. and Okumura, K. 2002. Effects of polymorphisms of MDR1, MRP1, and MRP2 genes on their mRNA expression levels in duodenal enterocytes of healthy Japanese. Biol Pharm Bull 25: 1356–1359.
Oselin K., Nowakowski-Gashaw I., Mrozikiewicz P.M., Wolbergs D., Pähkla R. and Roots I. 2003. Quantitative determination of MDR1 mRNA expression in peripheral blood lymphocytes: possible role of genetic polymorphisms in the MDR1 gene. Eur J Clin Invest 33: 261–267.
Owen, A., Chandler, B., Bray, P.G., Ward, S.A., Hart, C.A., Back, D.J. and Khoo, S.H. 2004. Functional correlation of P-glycoprotein expression and genotype with expression of human immunodeficiency virus type 1 coreceptor CXCR4. J Virol 78: 12022–12029.
Ozawa, S., Soyama, A., Saeki, M., Fukushima-Uesaka, H., Itoda, M., Koyano S., Sai, K., Ohno, Y., Saito, Y. and Sawada, J. 2004. Ethnic differences in genetic polymorphisms of CYP2D6, CYP2C19, CYP3As and MDR1/ABCB1. Drug Metab Pharmacokinet 19: 83–95.
Pauli-Magnus, C. and Kroetz, D.L. 2004. Functional implications of genetic polymorphisms in the multidrug resistance gene MDR1 (ABCB1). Pharm Res 21: 904–913.
Sakaeda, T., Nakamura, T., Horinouchi, M., Kakumoto, M., Ohmoto, N., Sakai, T., Morita, Y., Tamura, T., Aoyama, N., Hirai, M,, Kasuga, M. and Okumura, K. 2001. MDR1genotype-related pharmacokinetics of digoxin after single oral administration in healthy Japanese subjects. Pharm Res 18: 1400–1404.
Schaeffeler, E., Eichelbaum, M., Brinkmann, U., Penger, A., Asante-Poku, S., Zanger, U.M. and Schwab, M. 2001. Frequency of C3435T polymorphism of MDR1 gene in African people. Lancet 358: 383–384.
Schinkel, A.H. and Jonker, J.W. 2003. Mammalian drug efflux transporters of the ATP binding cassette (ABC) family: an overview. Adv Drug Deliv Rev 55: 3–29.
Sudchada, P., Oo-puthinan, S., Kerdpin, O. and Saelim N. 2010 ABCB1 gene expression in peripheral blood mononuclear cells in healthy Thai males and females. Genet Mol Res 9: 1177–1185.
Tanabe, M., Ieiri, I., Nagata, N., Inoue, K., Ito, S., Kanamori, Y., Takahashi, M., Kurata, Y., Kigawa, J., Higuchi, S., Terakawa, N. and Otsubo, K. 2001. Expression of P-glycoprotein in human placenta: relation to genetic polymorphism of the multidrug resistance (MDR)-1 gene. J Pharmacol Exp Ther 297: 1137–1143.
Tang, K., Ngoi, S.M., Gwee, P.C., Chua, J.M., Lee, E.J., Chong, S.S. and Lee C.G. 2002. Distinct haplotype profiles and strong linkage disequilibrium at the MDR1 multidrug transporter gene locus in three ethnic Asian populations. Pharmacogenetics 12: 437–450.
Yi, S.Y., Hong, K.S., Lim, H.S., Chung, J.Y., Oh, D.S., Kim, J.R. Jung, H.R., Cho, J.Y., Yu, K.S., Jang, I.J. and Shin, S.G. 2004. A variant 2677A allele of the MDR1 gene affects fexofenadine disposition. Clin Pharmacol Ther 76: 418–427.
Zhou, S.F. 2008. Structure, function and regulation of P-glycoprotein and its clinical relevance in drug disposition. Xenobiotica 38: 802–832.
