A Multiplex Polymerase Chain Reaction Protocol for the Simultaneous Analysis of the GlutathioneS-Transferase GSTM1 and GSTT1 Polymorphisms

Arand, Michael; Mühlbauer, R.; Hengstler, Jan G.; Jäger, Elke; Fuchs, Jürgen; Winkler, Lars; Oesch, Franz

doi:10.1006/abio.1996.0153

Cited by 291 publications

(166 citation statements)

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“…GSTM1 and GSTT1 null alleles were identified using a multiplex polymerase chain reaction (PCR)-based method (Arand et al 1996). The polymorphic site at nucleotide 638 in exon 7 (Arg213His (*2 allele), rs9282861) of the SULT1A1 gene was genotyped by PCR-restriction fragment length polymorphisms (RFLP) analysis as described by Coughtrie et al (Coughtrie et al 1999), CYP1A1 3 0 -flanking region MspI polymorphism (CYP1A1*2A allele, rs4646903), CYP2E1 PstI polymorphism [CYP2E1*5B allele, rs3813867 (PstI)] and CYP2E1 DraI (*5A or *6 alleles, rs6413432) were also determined by PCR-RFLP analyses.…”

Section: Dna Extraction and Genotypingmentioning

confidence: 99%

A case–control study on the effect of metabolic gene polymorphisms, nutrition, and their interaction on the risk of non-alcoholic fatty liver disease

et al. 2014

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The oxidative stress is a key issue in the etiology of non-alcoholic fatty liver disease (NAFLD). The aim of our study was to evaluate the effect of metabolic gene polymorphisms involved in the oxidative stress (GSTT1, GSTM1, SULT1A1, CYP2E1, and 1A1), lifestyle and nutrition aspects, and their interaction, on the risk of NAFLD. We enrolled 294 cases and 359 controls, and collected demographics, anthropometric, lifestyle, and nutrition data. A subgroup of NAFLD provided additional data on nutrients and on physical activity engagement. Each patient provided a blood sample for DNA extraction and genotyping. Clinical and laboratory data were collected from cases. Multivariable analysis shows a significant protective effect of age, gender, and moderate drinking habits on the risk of NAFLD, while an increased risk for greater consumption of fruit and grilled meat or fish. Significant interactions were reported between alcohol consumption, fruit intake, grilled meat and fish, and selected genetic variants. From the subgroup analysis, a moderate/high consumption of fat and/or grilled meat/fish, and a high consumption of white meat increase the risk of NAFLD. Engaging any physical activity at least 1 time/ week halves the risk of NAFLD. Besides confirming the beneficial effect of moderate alcohol intake and regular physical activity, and the increased risk associated with high fruit and fat intake, for the first time, we report a detrimental effect of grilled food on NAFLD risk. An effect modification by selected gene variants increases the risk in combination with fruit and grilled food intake.

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Section: Dna Extraction and Genotypingmentioning

confidence: 99%

A case–control study on the effect of metabolic gene polymorphisms, nutrition, and their interaction on the risk of non-alcoholic fatty liver disease

et al. 2014

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“…A PCR-based RFLP assay 27 was used to determine the SNP of SULT1A1. To genotype deletion polymorphisms in GSTM1 and GSTT1, a multiplex-PCR procedure was used, the primer sets being based on those described previously by Arand et al 28 Our genotyping protocol for the promoter region containing the TA repeats polymorphism of UGT1A1 was similar to that described previously, 23 using an ABI Prism 3100 DNA sequencer and GE-NESCAN3.1 and GENOTYPER 3.7 software. To ensure that the observed polymorphisms were specific and not the results of experimental variation, the results were confirmed by repeating 15% of the assays and by sequencing 10% of the specimens.…”

Section: Genotypic Polymorphism and Genotypingmentioning

confidence: 99%

Breast cancer risk associated with genotype polymorphism of the catechol estrogen‐metabolizing genes: A multigenic study on cancer susceptibility

Cheng

Chen

Huang

et al. 2004

Intl Journal of Cancer

102

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Estrogen has been suggested to trigger breast cancer development via an initiating mechanism involving its metabolite, catechol estrogen (CE). To examine this hypothesis, we carried out a multigenic case-control study of 469 incident breast cancer patients and 740 healthy controls to define the role of important genes involved in the different metabolic steps that protect against the potentially harmful effects of CE metabolism. We studied the 3 genes involved in CE detoxification by conjugation reactions involving methylation (catechol-O-methyltransferase, COMT), sulfation (sulfotransferase 1A1, SULT1A1), or glucuronidation (UDP-glucuronosyltransferase 1A1, UGT1A1), one (manganese superoxide dismutase, MnSOD) involved in protection against reactive oxidative species-mediated oxidation during the conversion of CEsemiquinone (CE-SQ) to CE-quinone (CE-Q), and 2 of the glutathione S-transferase superfamily, GSTM1 and GSTT1, involved in CE-Q metabolism. Support for this hypothesis came from the observations that (i) there was a trend toward an increased risk of breast cancer in women harboring a greater number of putative high-risk genotypes of these genes (p < 0.05); (ii) this association was stronger and more significant in those women who were more susceptible to estrogen [no history of pregnancy or older (>26 years) at first full-term pregnancy (FFTP)]; and (iii) the risks associated with having one or more high-risk genotypes were not the same in women having experienced different menarche-to-FFTP intervals, being more significant in women having been exposed to estrogen for a longer period (>12 years) before FFTP. Furthermore, because CE-Q can attack DNA, leading to the formation of double-strand breaks (DSB), we examined whether the relationship between cancer risk and the genotypic polymorphism of CE-metabolizing genes was modified by the genotypes of DSB repair genes, and found that a joint effect of CE-metabolizing genes and one of the two DSB repair pathways, the homologous recombination pathway, was significantly associated with breast cancer development. Based on comprehensive CE metabolizing gene profiles, our study provides support to the hypotheses that breast cancer can be initiated by estrogen exposure and that increased estrogen exposure confers a higher risk of breast cancer by causing DSB to DNA.Key words: breast cancer; estrogen; catechol estrogen; polymorphism; molecular epidemiology An increased risk of breast cancer due to prolonged exposure to estrogen has been well documented by epidemiological observations showing that estrogen-related risk factors, including age at menarche, age at menopause, parity and age at first full-term pregnancy (FFTP), are significantly associated with breast cancer risk. 1,2 Why estrogen exposure should increase breast cancer risk is an intriguing question that has not been conclusively answered. One simple explanation is that the proliferative effect of estrogen on breast epithelium promotes the growth of tumor cells, leading to progression of breast cancer. [3...

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“…Die Arbeitsersparnis bei einer gleichzeitigen Bestimmung von GSTT1-und GSTM1-Doppel-Null-Genotypen in einer Multiplex-PCR ist ebenso offensichtlich wie der Nutzen der Kontrollmöglichkeit für eine erfolgreiche Amplifikation durch einen internen Standard. Mehrere Arbeitsgruppen kombinierten die bekannten 459 bp und 218 bp großen GSTT1-und GSTM1-Ampflifikate in einer Reaktion [32,66,67]. Die Größe des Kontrollamplifikats wurde so gewählt, dass die zugehörige Elektrophoresebande zwischen denen der GST-Amplifikate liegt.…”

Section: Diskussion Der Methodeunclassified

“…Die Größe des Kontrollamplifikats wurde so gewählt, dass die zugehörige Elektrophoresebande zwischen denen der GST-Amplifikate liegt. Als interner Standard wurden Sequenzen des F-Aktin-Gens [66] bzw. des Exons 7 [67] oder des 3'-nicht-translatierten Bereichs [68] des CYP1A1-Gens für die PCR verwendet.…”

Section: Diskussion Der Methodeunclassified

Glutathion‐S‐Transferase T1 und M1 (Genotypisierung) [Biomonitoring Methods in German language, 2004]

Krause¹,

Müller²

2012

The MAK‐Collection for Occupational Health and Safety

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Veröffentlicht in der Reihe Analytische Methoden zur Prüfung gesundheitsschädlicher Arbeitsstoffe: Analysen in biologischem Material , 16. Lieferung, Ausgabe 2004 Der Artikel enthält folgende Kapitel: Grundlage des Verfahrens Geräte, Chemikalien, Lösungen, Gele Geräte Chemikalien Lösungen PCR‐Mastermix für 20 Proben in 50‐µL‐Ansätzen Gele Probenaufarbeitung Lagerung DNA‐Isolierung Bestimmung der Reinheit und Konzentration der DNA Vorbereitung und Durchführung der PCR PCR Elektrophorese Kalibrierung Auswertung Qualitätskontrolle Störeinflüsse Diskussion der Methode Addendum Zusätzlich benötigte Reagenzien und Geräte: Mastermixes Restriktionsenzyme Amplifikation unter Verwendung von PCR‐Beads Restriktionsanalyse der PCR‐Produkte Gelelektrophoretische Trennung Auswertung

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A Multiplex Polymerase Chain Reaction Protocol for the Simultaneous Analysis of the GlutathioneS-Transferase GSTM1 and GSTT1 Polymorphisms

Cited by 291 publications

References 1 publication

A case–control study on the effect of metabolic gene polymorphisms, nutrition, and their interaction on the risk of non-alcoholic fatty liver disease

A case–control study on the effect of metabolic gene polymorphisms, nutrition, and their interaction on the risk of non-alcoholic fatty liver disease

Breast cancer risk associated with genotype polymorphism of the catechol estrogen‐metabolizing genes: A multigenic study on cancer susceptibility

Glutathion‐S‐Transferase T1 und M1 (Genotypisierung) [Biomonitoring Methods in German language, 2004]

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