Interactions between GSTM1, GSTT1 and GSTP1 polymorphisms and smoking and intake of fruit and vegetables in relation to lung cancer

doi:10.1016/j.lungcan.2006.10.010

Lung Cancer

Volume 55, Issue 2, February 2007, Pages 137-144

https://doi.org/10.1016/j.lungcan.2006.10.010 Get rights and content

Summary

Polymorphisms in glutathione S-transferases (GST) are weakly associated with risk for lung cancer. We examined gene–environment interactions in relation to lung cancer in 430 cases and 767 comparison persons identified within a prospective cohort of 57,053 persons. We used assays capable of discriminating heterozygous individuals from those with two functional alleles and homozygous deletions in GSTM1 and GSTT1. There was no overall association between the GST polymorphisms and lung cancer. We found that fruit and vegetables reduced the risk of lung cancer only among carriers of at least one functional GSTM1 allele, and among carriers of two GSTP1 Val alleles. There were no significant interactions between the GST polymorphisms and smoking.

Introduction

Tobacco smoking causes the vast majority of lung cancer cases. Glutathione S-transferase (GST) enzymes metabolise potential carcinogens present in tobacco smoke through a conjugation with reduced glutathione. GSTs are a multigene family of phase II enzymes. GSTM1 and GSTP1 metabolise large hydrophobic electrophiles, such as polycyclic aromatic hydrocarbons derived epoxides [1], whereas GSTT1 is involved in the metabolism of smaller compounds, such as monohalomethane and ethylene oxide [2]. GSTs also metabolise compounds formed during oxidative stress, such as hydroperoxides and oxidized lipids, and they are transcriptionally activated during oxidative stress [3].

A homozygous gene deletion that results in total lack of the enzyme (null genotype) is a common polymorphism in both the GSTM1 and the GSTT1 gene. Most previous studies have compared individuals with at least one functional allele (+/+ or +/0) with individuals without any functional alleles (0/0). Meta-analyses have indicated that carriers of GSTM1 0/0 or GSTT1 0/0 have a slightly higher risk of lung cancer as compared with carriers of at least one functional allele [4], [5]. An A/G single nucleotide polymorphism (SNP) located within the substrate-binding domain of GSTP1 results in an amino acid substitution of isoleucine by valine (Ile¹⁰⁵Val). This polymorphism influences enzyme activity [6], but epidemiological studies of the possible relationship between the GSTP1 polymorphism and lung cancer show inconsistent results [7], [8].

Recently, a quantitative PCR-based method has made it possible to discriminate subjects carrying zero, one or two functional alleles of the GSTM1 or the GSTT1 gene [9]. Two studies have investigated the associations between the three-genotype determination of GSTM1 and GSTT1 and colorectal cancer and bladder cancer, respectively, and they indicated that phenotypic differences between +/+ and +/0 genotypes exist [10], [11]. No studies have investigated this in relation to lung cancer.

The purpose of this study was to investigate the relationship between polymorphisms in GSTM1, GSTT1 and GSTP1 and risk of lung cancer, and interactions between the polymorphisms and intake of fruit and vegetables and smoking in the development of lung cancer.

Section snippets

Study group

Diet, Cancer and Health (DCH) is a Danish prospective follow-up study. Invited to participate were 160,725 individuals aged 50–64 years of which 57,053 individuals with no previous cancer diagnosis were recruited [12]. At enrolment (1993–1997), detailed information on diet and smoking habits were collected, and blood was sampled and stored at −150 °C. All participants gave informed consent. The Danish Ethical Committee approved the study.

Data on diet intake was obtained from a 192-item,

Results

The proportion of current smokers and the smoking intensity was substantially higher, and the duration of smoking was substantially longer, among cases than among the sub-cohort members, and the median dietary intake of fruit, all vegetables and cruciferous vegetables was lower among cases than among sub-cohort members (Table 1). Intake of fruit was correlated with intake of total vegetables (Rs = 0.41, P < 0.0001) and intake of cruciferous vegetables (Rs = 0.29, P < 0.0001). Intake of total vegetables

Discussion

There was no overall effect of either of the GST polymorphisms on risk for lung cancer. There were significant interactions between the GSTM1 polymorphism and intake of fruits and vegetables and between the GSTP1 polymorphism and intake of vegetables in relation to risk of lung cancer.

The present case–cohort study is population-based and both cases and their comparison group were selected from the same cohort, which together with complete follow-up of the participants minimizes the risk for

Conflict of interest

None declared.

Acknowledgement

We thank Karsten Sørensen for excellent technical assistance.

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Cited by (34)

Genetic polymorphisms in GSTM1, GSTT1 and GSTP1 genes and risk of lung cancer in a North Indian population
2015, Cancer Epidemiology
Citation Excerpt :
Individuals carrying both null genotypes for GSTM1 and GSTT1 genes had an increased risk for lung cancer as compared to subjects having both genes. These findings are in concurrence with those of previous studies [24,59–61]. However, some reports have failed to find such an association in other populations [13,27,46].
A number of studies done so far in different populations have shown that polymorphisms within the GST genes play an important role in determining individual susceptibility to lung cancer; however, data obtained so far have been contradictory within the same or different populations. Few studies have focused on the combinatorial effect of the GST genes on susceptibility to lung cancer and also for different histological subtypes. Our aim is to investigate the roles of GSTM1, GSTT1, and GSTP1 polymorphisms as genetic modifiers of risk for lung cancer and histological subtypes using a larger sample size in a North Indian population.
In total 540 subjects (270 lung cancer cases and 270 controls) were evaluated for the GST polymorphism. Genotyping for the GSTM1, GSTT1 and GSTP1 gene was done by using a multiplex PCR and PCR-RFLP method.
GSTM1 null genotype was found to be associated with lung cancer (OR = 1.65, 95%CI: 116–2.3, P = 0.005) and this risk was higher in cases of adenocarcinoma (ADCC). GSTT1 and GSTP1 did not show any significant association with lung cancer; however, when stratified for histological subtypes a significant association was observed for ADCC and small-cell lung cancer (SCLC) for both GSTT1 null and variant GSTP1 genotypes. The combined ‘at risk’ genotypes of null GSTM1 and GSTT1 genes were found to be associated with lung cancer risk, and this risk was higher in cases of ADCC (OR = 4.09, 95%CI: 110–10.2, P = 0.002). There is a twofold increased risk for lung cancer with the null GSTM1 and wild-type GSTP1 genotypes (P = 0.0004); similarly, a fourfold increased risk was observed with the null GSTT1 and wild-type GSTP1 genotypes (P = 0.0001).
The deficient GST genotypes seem thus to be important risk modifiers for lung cancer and related histological subtypes, especially in combination.
Genetic polymorphism of glutathione S-transferase P1 (GSTP1) in Delhi population and comparison with other global populations
2014, Meta Gene
Glutathione S-transferases (GSTs) belong to a super family of phase II detoxification enzymes, which play an important role in protecting cells from damage caused by endogenous and exogenous compounds by conjugating reactive intermediates with glutathione to produce less reactive water-soluble compounds. In the present study, we determined the frequencies of two polymorphisms in exon 5 and exon 6 of GSTP1 gene in 500 normal individuals from Delhi. GSTP1 polymorphism was analysed by PCR-RFLP using amplification refractory mutation system (ARMS) assay. Two polymorphic sites in GSTP1 (Ile105 → Val105; Ala114 → Val114) have been analysed simultaneously, which results in four alleles: GSTP1*A (wild-type Ile105; Ala114), GSTP1*B (Val105; Ala114), GSTP1*C (Val105; Val114) and GSTP1*D (Ile105; Val114). The GSTP1 allele frequency in Delhi population was 0.663, 0.248, 0.069, and 0.020 for GSTP1*A, GSTP1*B, GSTP1*C, and GSTP1*D respectively. The frequency of Ile105 and Val105 allele was 0.683 and 0.317 respectively and it was calculated for the purpose of comparison with published data where all the four alleles were not analysed. GSTP1 alleles from Delhi population were compared with reported frequencies from all over India, and from other ethnic groups worldwide. This study would provide a basic database for future genetic studies.
Candidate dietary phytochemicals modulate expression of phase II enzymes GSTP1 and NQO1 in human lung cells
2010, Journal of Nutrition
Many phytochemicals possess cancer-preventive properties, some putatively through phase II metabolism-mediated mutagen/oxidant quenching. We applied human lung cells in vitro to investigate the effects of several candidate phytopreventive agents, including green tea extracts (GTE), broccoli sprout extracts (BSE), epigallocatechin gallate (EGCG), sulforaphane (SFN), phenethyl isothiocyanate (PEITC), and benzyl isothiocyanate (BITC), on inducing phase II enzymes glutathione S-transferase P1 (GSTP1) and NAD(P)H:quinone oxidoreductase 1 (NQO1) at mRNA and protein levels. Primary normal human bronchial epithelial cells (NHBE), immortalized human bronchial epithelial cells (HBEC), and lung adenocarcinoma cells (A549) were exposed to diet-achievable levels of GTE and BSE (0.5, 1.0, 2.0 mg/L), or individual index components EGCG, SFN, PEITC, BITC (0.5, 1.0, 2.0 μmol/L) for 24 h, 48 h, and 6 d, respectively. mRNA assays employed RNA-specific quantitative RT-PCR and protein assays employed Western blotting. We found that in NHBE cells, while GSTP1 mRNA levels were slightly but significantly increased after exposure to GTE or BSE, NQO1 mRNA increased to 2- to 4-fold that of control when exposed to GTE, BSE, or SFN. Effects on NQO1 mRNA expression in HBEC cells were similar. NQO1 protein expression increased up to 11.8-fold in SFN-treated NHBE cells. Both GSTP1 and NQO1 protein expression in A549 cells were constitutively high but not induced under any condition. Our results suggest that NQO1 is more responsive to the studied chemopreventive agents than GSTP1 in human lung cells and there is discordance between single agent and complex mixture effects. We conclude that modulation of lung cell phase II metabolism by chemopreventive agents requires cell- and agent-specific discovery and testing.
Copy Number Variants of GSTM1 and GSTT1 in Relation to Lung Cancer Risk in a Prospective Cohort Study
2009, Annals of Epidemiology
Citation Excerpt :
These results are consistent with previous observation that hemizygous and wild-type genotypes are associated with functional differences in enzymatic activity (15). The only previous study we are aware of to report on the associations between GSTM1 and GSTT1 genotypes and lung cancer risk that classified hemizygous deletion separately from wild-type observed no difference between the hemizygous and homozygous null genotypes of GSTM1 and GSTT1 when compared to the referent wild-type genotype (34). Our results are in agreement with the previous lack of associations observed for GSTT1, and thus suggest caution in interpreting our results for GSTM1.
Previous studies did not discriminate wild-type from hemizygous genotypes of GSTM1 and GSTT1. In this study, we investigated wild-type, hemizygous deletion, and homozygous deletion genotypes of GSTM1 and GSTT1 and lung cancer risk.
We conducted a nested case-control study of 143 primary incident lung cancer cases matched to 447 cancer-free controls. Genotyping was carried out using a real-time polymerase chain reaction (PCR)-based assay. Conditional logistic regression models were used to estimate odds ratios (OR) and 95% confidence intervals (CI).
Compared to GSTM1 wild-type carriers, the relative odds of lung cancer increased from 1.49 (95% CI = 0.66–3.40) to 1.80 (95% CI = 0.81–4.02) for the hemizygous and homozygous deletion genotypes, respectively (p-trend = 0.13). The strongest associations were seen among those who smoked less than one pack per day and had greater than or equal to one deletion variant of GSTM1 (OR = 3.25; 95% CI = 0.93–11.34; p-trend = 0.07) whereas the reverse was observed for smokers who smoked greater than or equal to one pack per day (OR = 0.80; 95% CI = 0.24–2.67; p-interaction = 0.08). No clear associations were observed for GSTT1 genotypes.
Risk of lung cancer increased as the number of deletion variants increased for GSTM1, although the associations were nonsignificant. Discriminating between the wild-type, hemizygous, and homozygous deletion GSTM1 genotypes permitted a more precise characterization of the associations between GSTM1 deletion variants and lung cancer.
GSTM1 copy number and lung cancer risk
2009, Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis
The GSTM1 null genotype is associated with a small increased lung cancer risk when compared to controls with at least one copy of the GSTM1 gene. As two copies of the GSTM1 gene might provide more protection than a single copy, we have determined GSTM1 copy number in a lung cancer case-control study. Cases with incident lung cancer were identified through a Bronchoscopy Unit and two separate hospital based control groups with non-malignant disease were selected with one from the same Bronchoscopy Unit and the other from a chest clinic at the same hospital. Subjects with at least one GSTM1 copy had a decreased lung cancer risk whatever the control group: the odds ratio (95% CI), after adjustment for age, gender and smoking duration, was 0.64 (0.41–0.98) and 0.54 (0.32–0.91) with bronchoscopy and chest clinic controls, respectively. Lung cancer risk varied with GSTM1 copy number with chest clinic controls only: the OR was 0.56 (0.32–0.97) for one copy of the GSTM1 gene and with two copies 0.43 (0.15–1.22), a trend that was significant (p = 0.02): with bronchoscopy controls the trend was not significant (p = 0.07). Results then confirm that the presence of GSTM1 provides protection against the risk of lung cancer. In addition there is equivocal evidence that this protection varies with the number of gene copies.
Haplotype-tagging single nucleotide polymorphisms in the GSTP1 gene promoter and susceptibility to lung cancer
2009, Cancer Epidemiology
Background: Glutathione S-transferase (GST) P1 is a major phase II xenobiotic-metabolizing enzyme in the human lung. Our laboratory had previously identified nine single nucleotide polymorphisms (SNPs) in the GSTP1 gene promoter, which were then grouped into three main haplotypes (Hap1, Hap2, and Hap3) based on statistical inference. Hap3 was found to display a high expression phenotype. The main objective of the current study was to test the association between GSTP1 promoter haplotypes with the risk of lung cancer after determining the promoter haplotypes experimentally through cloning and sequencing. Methods: We conducted a case–control analysis of 150 subjects with lung cancer and 329 controls with no personal history of the disease. The three statistically inferred GSTP1 promoter haplotypes were confirmed experimentally through cloning and sequencing. Haplotype-tagging SNPs were selected and GSTP1 haplotypes were tested for genetic association to lung cancer using unconditional logistic regression after adjusting for confounders. Statistical interaction between GSTP1 promoter haplotypes with either cigarette smoking or dietary fruit and vegetable intake were tested using the likelihood ratio test. Results: We did not find protective effects of Hap3 against lung cancer, despite an adequately powered design for this main effect. Homozygous variants of tagSNPs −1738 T > A and −354 G > T, which tag Hap2, showed an increased (but statistically non-significant) risk of lung cancer among all subjects as well as among individuals with low fruit and vegetable intake, compared to homozygous wildtypes for these SNPs. We did not find significant interactions between Hap2 and dietary intake of fruits and vegetables. Conclusions: Our results do not support significant main and modifying effects for GSTP1 promoter haplotypes on susceptibility to lung cancer in this population, but reinforce the protective effects of dietary intake of fruits and vegetables.

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Interactions between GSTM1, GSTT1 and GSTP1 polymorphisms and smoking and intake of fruit and vegetables in relation to lung cancer

Summary

Introduction

Section snippets

Study group

Results

Discussion

Conflict of interest

Acknowledgement

Mutat Res

J Biol Chem

Cancer Lett

Lancet

J Clin Epidemiol

Lung Cancer

Lung Cancer

Lung Cancer

J Nutr

Lancet

Lancet

The glutathione S-transferase supergene family: regulation of GST and the contribution of the isoenzymes to cancer chemoprotection and drug resistance

Crit Rev Biochem Mol Biol

Glutathione transferases

Annu Rev Pharmacol Toxicol

Meta- and pooled analyses of the effects of glutathione S-transferase M1 polymorphisms and smoking on lung cancer risk

Carcinogenesis

Combinations of glutathione S-transferase genotypes and risk of early-onset lung cancer in Caucasians and African Americans: a population-based study

Carcinogenesis

Possible gene dosage effect of glutathione S-transferases on atopic asthma: using real-time PCR for quantification of GSTM1 and GSTT1 gene copy numbers

Hum Mutat

GSTM1, GSTT1, and GSTP1 polymorphisms and risk of advanced colorectal adenoma

Cancer Epidemiol Biomarkers Prev

Diet, cancer and health—a population study and establishment of a biological bank in Denmark

Ugeskr Laeger

Development of a semiquantitative food frequency questionnaire to assess food, energy and nutrient intake in Denmark

Int J Epidemiol