Aberrant DNA methylation of drug metabolism and transport genes in nodular goiter
© Zhang et al; licensee BioMed Central Ltd. 2011
Received: 31 August 2011
Accepted: 12 October 2011
Published: 12 October 2011
The genes encoding drug-metabolizing enzymes and transporters play an important role in maintaining the normal life processes of human body. Their disorder or defect will lead to the occurrence and development of various diseases. Currently, most of studies have focused on genetic variations in these genes, however, in the present study, we analyzed promoter methylation of 11 drug metabolism and transport genes in a cohort of nodular goiter and normal thyroid tissues using methylation-specific PCR (MSP). Our data first revealed a distinct methylation profiling in drug metabolism and transport genes between nodular goiter and normal thyroid tissues, particularly ABCB4, CYP1B1 and CYP24A1 and SLC1A2. Given these genes contribute to the development and progression of various diseases, such as multidrug resistance and tumorigenesis, these epigenetic events may thus play a critical role in the pathogenesis of nodular goiter.
The genes encoding drug-metabolizing enzymes and transporters play an important role in transporting various kinds of molecules import or export the cells, which is closely associated with the development of various human diseases, mainly including solute carrier (SLC) superfamily, cytochrome P450 (CYP) superfamily and ATP binding cassette (ABC) superfamily . To date, most of studies focused on investigating SNPs or gene mutation in these genes, however, it is has recently been reported that epigenetic mechanisms were involved in the regulation of these genes . In the present study, we choose 11 drug metabolism and transport genes, including ABCB1, ABCB4, ABCG2, CYP1A1, CYP1B1, CYP24A1, CYP27B1, CYP39A1, SLC1A2, SLC19A3, and SLC26A2, to detect their methylation status of promoter region in a cohort of nodular goiter and normal thyroid tissues using methylation-specific PCR (MSP).
Methylation-specific PCR (MSP) primers used in the present study
Annealing temperature (°C)
To date, most of studies have focused on polymorphisms (e.g. SNPs) or gene mutation in drug metabolism and transport genes in some human diseases, including thyroid disease, such as Graves' disease . Unlike the previous studies, this study systematically analyzed promoter methylation of drug metabolism and transport genes in nodular goiter tissues. Promoter methylation is the most common epigenetic events that aberrantly regulate gene expression in different tissues, which play a key role in the early disease . The pathogenesis of nodular goiter is multifactorial and probably differs from patient to patient. In contrast to the endemic goiter, iodine deficiency is not a primary causal factor. Environmental factors, such as natural goitrogens, iodine intake, malnutrition, drugs, stress, pollution or infections, constitutional factors, such as female gender, and several genetic factors, such as circulating thyroid growth factors, contribute to different degree to the development of nodular thyroid enlargement. However, in the present study, our data suggested that aberrant methylation of some genes, such as drug metabolism and transport genes, may play an important role in the pathogenesis of nodular goiter. In support of this, a previous study also showed that the Platelet-derived growth factor B-chain was also abnormally methylated in multinodular goiters .
ABCB4 belongs to the multidrug resistance subfamily, encoding a full transporter and member of the p-glycoprotein family of membrane proteins with phosphatidylcholine as its substrate , which was more frequently methylated in normal thyroid tissue compared with nodular goiter in the present study. Of note, this gene was highly expressed in various human cancers [8, 9]. Its overexpression may induce multidrug resistance and create a waste of energy . CYP1B1 belongs to the cytochrome P450 family 1 and is a major member of the extrahepatic xenobiotic-metabolizing CYP enzyme family. It can transform estrogen to strong carcinogenic compounds 4-hydroxy estrogen, and is overexpressed in various malignant tumors, and promotes the development of tumors [10, 11]. CYP1B1 was significantly lowly methylated in nodular goiter compared with normal thyroid tissues in the present study, suggesting that its aberrant expression may play a key role in in the pathogenesis of nodular goiter. CYP24A1 is relevant with the metabolism of vitamin D3, which emerged as a protective factor for human body. CYP24A1 encodes a mitochondrial protein which initiates the degradation of 1,25-dihydroxyvitamin D3, the physiologically active form of vitamin D3, by hydroxylation of the side chain . A previous study showed that the administration of calcitriol in combination with CYP24A1 inhibitor enhanced antiproliferative effects, increased systemic calcitriol exposure, and promoted the activation of caspase-independent apoptosis pathway . Interestingly, a very recent study showed that that CYP24A1 expression is marked increased in papillary thyroid cancer (PTC) compared with normal thyroid tissues . It is consistent with our recent findings that CYP24A1 was significantly lowly methylated in PTC compared with normal tissues (Lihong Zhang, Jing Shi, Meiju Ji, Guanjun Zhang, Jiao Fu, Li Xu, Bingyin Shi, and Peng Hou: Methylation analysis of drug metabolism and transport genes in papillary thyroid cancer, submitted). SLC1A2 encodes a membrane-bound protein which is the principal transporter that clears the excitatory neurotransmitter glutamate from the extracellular space at synapses in the central nervous system, to prevent neuronal damage from excessive activation of glutamate receptors . Importantly, our data showed that demethylation agent 5-Aza-2'-dC could induce the expression of these methylated genes in at least one cell line, indicating that expression of these genes could be regulated by promoter methylation (data not shown).
In conclusions, although nodular goiter is a benign disease, it seriously influences the quality of life of many patients because it usually causes dyspnea and swallowing difficulties. Thus, it is amazing if we can inhibit the occurrence or proceeding of this disease. In the present study, we observed that a distinct methylation profiling in drug metabolism and transport genes between nodular goiter and normal thyroid tissues, suggesting that aberrant expression of these genes may play an important role in the pathogenesis of nodular goiter.
This work was supported by the National Natural Science Foundation of China (No. 30901459 and 30973372).
- Lee NH: Pharmacogenetics of drug metabolizing enzymes and transporters: effects on pharmacokinetics and pharmacodynamics of anticancer agents. Anticancer Agents Med Chem 2010, 10: 583–592. 10.2174/187152010794474019PubMed CentralView ArticlePubMedGoogle Scholar
- Hirota T, Takane H, Higuchi S, Ieiri I: Epigenetic regulation of genes encoding drug-metabolizing enzymes and transporters; DNA methylation and other mechanisms. Curr Drug Metab 2008, 9: 34–38. 10.2174/138920008783331130View ArticlePubMedGoogle Scholar
- Hou P, Ji M, Xing M: Association of PTEN gene methylation with genetic alterations in the phosphatidylinositol 3-kinase/AKT signaling pathway in thyroid tumors. Cancer 2008, 113: 2440–2447. 10.1002/cncr.23869View ArticlePubMedGoogle Scholar
- Bufalo NE, Santos RB, Cury AN, Andrade RA, Morari J, Morari EC, Leite JL, Monte O, Romaldini JH, Ward LS: Genetic polymorphisms associated with cigarette smoking and the risk of Graves' disease. Clin Endocrinol (Oxf) 2008, 68: 982–987. 10.1111/j.1365-2265.2007.03121.xView ArticleGoogle Scholar
- Moss TJ, Wallrath LL: Connections between epigenetic gene silencing and human disease. Mutat Res 2007, 618: 163–174. 10.1016/j.mrfmmm.2006.05.038PubMed CentralView ArticlePubMedGoogle Scholar
- Matsuo K, Tang SH, Zeki K, Gutman RA, Fagin JA: Aberrant deoxyribonucleic acid methylation in human thyroid tumors. J Clin Endocrinol Metab 1993, 77: 991–995. 10.1210/jc.77.4.991PubMedGoogle Scholar
- Oude Elferink RP, Paulusma CC: Function and pathophysiological importance of ABCB4 (MDR3 P-glycoprotein). Pflugers Arch 2007, 453: 601–610. 10.1007/s00424-006-0062-9View ArticlePubMedGoogle Scholar
- Arai Y, Masuda M, Sugawara I, Arai T, Motoji T, Tsuruo T, Oshimi K, Mizoguchi H: Expression of the MDR1 and MDR3 gene products in acute and chronic leukemias. Leuk Res 1997, 21: 313–319. 10.1016/S0145-2126(96)00130-0View ArticlePubMedGoogle Scholar
- Meijer GA, Schroeijers AB, Flens MJ, Meuwissen SG, van der Valk P, Baak JP, Scheper RJ: Increased expression of multidrug resistance related proteins Pgp, MRP1, and LRP/MVP occurs early in colorectal carcinogenesis. J Clin Pathol 1999, 52: 450–454. 10.1136/jcp.52.6.450PubMed CentralView ArticlePubMedGoogle Scholar
- Saini S, Hirata H, Majid S, Dahiya R: Functional significance of cytochrome P450 1B1 in endometrial carcinogenesis. Cancer Res 2009, 69: 7038–7045. 10.1158/0008-5472.CAN-09-1691View ArticlePubMedGoogle Scholar
- Su JM, Lin P, Wang CK, Chang H: Overexpression of cytochrome P450 1B1 in advanced non-small cell lung cancer: a potential therapeutic target. Anticancer Res 2009, 29: 509–515.PubMedGoogle Scholar
- Sakaki T, Kagawa N, Yamamoto K, Inouye K: Metabolism of vitamin D3 by cytochromes P450. Front Biosci 2005, 10: 119–134.PubMedGoogle Scholar
- Muindi JR, Yu WD, Ma Y, Engler KL, Kong RX, Trump DL, Johnson CS: CYP24A1 inhibition enhances the antitumor activity of calcitriol. Endocrinology 2010, 151: 4301–4312. 10.1210/en.2009-1156PubMed CentralView ArticlePubMedGoogle Scholar
- Balla B, Kósa JP, Tobiás B, Halászlaki C, Takács I, Horváth H, Speer G, Nagy Z, Horányi J, Járay B, Székely E, Lakatos P: Marked increase in CYP24A1 gene expression in human papillary thyroid cancer. Thyroid 2011, 21: 459–460. 10.1089/thy.2010.0420View ArticlePubMedGoogle Scholar
- Levy LM, Warr O, Attwell D: Stoichiometry of the glial glutamate transporter GLT-1 expressed inducibly in a Chinese hamster ovary cell line selected for low endogenous Na+-dependent glutamate uptake. J Neurosci 1998, 18: 9620–9628.PubMedGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.