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Carcinogenesis Advance Access originally published online on June 8, 2007
Carcinogenesis 2007 28(8):1788-1793; doi:10.1093/carcin/bgm132
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Published by Oxford University Press 2007.

Evaluation of genetic variation in the double-strand break repair pathway and bladder cancer risk

Jonine D. Figueroa1,*, Núria Malats2, Nathaniel Rothman1, Francisco X. Real3, Debra Silverman1, Manolis Kogevinas2,5, Stephen Chanock1,6,10, Meredith Yeager6, Robert Welch6, Mustafa Dosemeci1, Adonina Tardón7, Consol Serra4, Alfredo Carrato8, Reina García-Closas9, Gemma Castaño-Vinyals2 and Montserrat García-Closas1

1 Division of Cancer Epidemiology and Genetics, National Cancer Institute, Department of Health and Human Services, Bethesda, MD, USA
2 Center for Research in Environmental Epidemiology, Barcelona, Spain
3 Unitat de Biologia Cellular i Molecular, Institut Municipal d'Investigació Mèdica, Barcelona, Spain
4 Universitat Pompeu Fabra, Barcelona, Spain
5 Medical School, Heraklion, Greece
6 Core Genotype Facility at the Advanced Technology Center of the National Cancer Institute, Gaithersburg, MD, USA
7 Universidad de Oviedo, Oviedo, Spain
8 Medical Oncology Department, Elche University Hospital, Elche, Spain
9 Unidad de Investigación, Hospital Universitario de Canarias, La Laguna, Spain
10 Pediatric Oncology Branch, National Institutes of Health, Bethesda, MD, USA

* To whom correspondence should be addressed. Tel: 301 402 3654; Fax: 301 402 0916; Email: figueroaj{at}mail.nih.gov


   Abstract
Top
 Abstract
Introduction
 Materials and methods
 Results
 Discussion
 Funding
 Supplementary material
 References
 
The double-strand break DNA repair (DSBR) pathway is implicated in maintaining genomic stability and therefore could affect bladder cancer risk. Here we present data evaluating 39 single-nucleotide polymorphisms (SNPs) in seven candidate genes whose products are involved in DNA break sensing (NBS1, BRCA1 interacting genes BRIP1 and ZNF350), non-homologous end-joining (NHEJ) DNA repair (XRCC4) and homologous recombination (HR) repair (RAD51, XRCC2 and XRCC3). SNPs for RAD51 and XRCC2 covered most of the common variation. Associations with bladder cancer risk were evaluated in 1150 newly diagnosed cases of urinary bladder transitional cell carcinomas and 1149 controls conducted in Spain during 1997–2001. We found that the genetic variants evaluated significantly contributed to bladder cancer risk (global likelihood ratio test P = 0.01). Subjects with the ZNF350 R501S (rs2278415) variant allele showed significantly reduced risk compared with common homozygote variants, odds ratio (OR) [95% confidence interval (95% CI)]: 0.76 (0.62–0.93) per variant allele. Carriers of a putative functional SNP in intron 7 of XRCC4 (rs1805377) had significantly increased bladder cancer risk compared with common homozygotes: 1.33 (1.08–1.64) per variant allele. Lastly, XRCC2 homozygote variants for three promoter SNPs (rs10234749, rs6464268, rs3218373) and one non-synonymous SNP (rs3218536, R188H) were associated with reduced bladder cancer risk (ORs ranging from 0.36 to 0.50 compared with common homozygotes). Meta-analysis for XRCC3 T241M (rs861539) had a significant small increase in risk among homozygote variants: OR (95% CI) = 1.17 (1.00–1.36). Results from this study provide evidence for associations between variants in genes in the DSBR pathway and bladder cancers risk that warrant replication in other study populations.

Abbreviations: CI, confidence interval; DSB, double-strand break; DSBR, double-strand break DNA repair; FDR, false discovery rate; HR, homologous recombination; LRT, likelihood ratio test; NHEJ, non-homologous end joining; OR, odds ratio; SNP, single-nucleotide polymorphism


   Introduction
Top
 Abstract
 Introduction
Materials and methods
 Results
 Discussion
 Funding
 Supplementary material
 References
 
Double-strand break DNA repair (DSBR) is implicated in maintaining genomic stability and cancer susceptibility. DNA double-strand breaks (DSBs) can promote genomic instability resulting in chromosomal abnormalities (1–4), which can arise from a variety of exogenous and endogenous exposures including ionizing radiation and tobacco smoke (4–7). Given the importance of DSBR in cellular genomic maintenance, inter-individual variation in DSBR pathway genes that sense and repair this damage may contribute to bladder cancer risk.

Repair of DSBs is a complex process. The first response is sensing a DSB that is mediated by the NBS1 gene and ATM protein kinase (5,8). NBS1–ATM sensing complex has been shown to interact with BRCA1 that has been proposed as a scaffolding protein (3). In addition, BRCA1 has been implicated in DNA damage signaling and can interact with other proteins including the transcriptional repressor ZNF350 (also known as ZBRK1). ZNF350-binding sequences are located at the regulatory region of many DNA damage-inducible genes and it has been shown that ZNF350 is degraded upon response to DNA damage in a BRCA1-dependent manner (9,10). BRIP1 (also known as BACH1) is a DNA helicase and BRCA1-interacting protein that has been implicated in the performance of BRCA1's DNA repair and its tumor suppressor function (11,12).

Once DSBs are recognized, cells can repair a break lesion in one of two ways: homologous recombination (HR) or non-homologous end joining (NHEJ). NHEJ does not rely on homologous sequences for template copy and involves recognition of the break and subsequent ligation by XRCC4 and ligase IV. NHEJ is often accompanied by the loss of genomic material in the repaired DNA molecule (5,8). Unlike NHEJ, repair by HR involves copying information from a homologous template and does not result in the loss of genetic information. HR encompasses many genes, but major players include RAD51 and RAD51-like genes such as XRCC2 and XRCC3 (5,8).

Recent evidence suggest that DSBs could be relevant to bladder cancer, including significant excess risk among populations exposed to ionizing radiation which cause DSBs (13,14); reports of somatic mutations and altered expression of DNA damage response pathway genes ATM and CHK2 (15) and DSBR NHEJ mechanisms to be more error prone in bladder tumors (16). We hypothesized that common genetic variation in DSBR genes may alter repair function and be associated with bladder cancer risk. Therefore, we evaluated 39 single-nucleotide polymorphisms (SNPs) in seven candidate genes whose products are involved in DNA break sensing (NBS1 and BRCA1 interacting genes BRIP1 and ZNF350), NHEJ DNA repair (XRCC4) and HR repair (RAD51, XRCC2 and XRCC3), in 1150 cases of urinary bladder transitional cell carcinomas and 1149 controls from the Spanish Bladder Cancer Study.


   Materials and methods
Top
 Abstract
 Introduction
 Materials and methods
Results
 Discussion
 Funding
 Supplementary material
 References
 
Study population
The Spanish Bladder Cancer Study design has been described elsewhere (17,18). In brief, cases were patients with a new diagnosis of histologically confirmed carcinoma of the urinary bladder from 1998 to 2001, aged 21–80 years [mean (SD) = 66 (10) years], of which 87% were males. Controls were selected from patients admitted to participating hospitals for diagnoses thought to be unrelated to the exposures of interest, individually matched to the cases for age in 5 year categories, sex, ethnicity and region. Overall, 84% (n = 1219) of eligible cases and 88% (n = 1271) of eligible controls agreed to participate in the study and were interviewed. Most of these subjects (97% of cases and 92% of controls) provided a blood or buccal cell sample as a source of DNA. The final study population available for analysis included 1150 cases and 1149 controls after exclusions for low DNA yield or quality, missing smoking information, non-transitional urothelial tumors and non-Caucasian descent.

Subjects were categorized as never smokers (29% of controls) if they smoked <100 cigarettes in their lifetime, and ever smokers otherwise. Ever smokers were further classified as regular smokers (63% of controls) if they smoked one cigarette per day for 6 months or longer, and occasional smokers (8% of controls) otherwise. Of controls that were regular smokers, 37% were current smokers (i.e. they smoked within a year of the reference date) and 63% were former smokers.

Genotyping
Genotyping was performed using three different methods:

TaqMan (Applied Biosystems, Foster City, CA) assays were used to genotype genomic DNA for three SNPs (rs3218536, rs861539, rs1805794). SNP selection favored SNPs with expected minor allele frequency >0.05 in Caucasians, non-synonymous SNPs, those previously evaluated in relation to bladder cancer risk or those with evidence of functional significance.
GoldenGate (Illumina®, San Diego, CA) assay was used to genotype 26 other SNPs (19) (Table I). SNP selection criteria were similar to those used for TaqMan assays, with the exception of RAD51 where SNPs chosen covered 89% of common variation, according to HapMap for Utah residents with ancestry from northern and western Europe (CEU) (20). Sixty-four (5.6%) of the 1150 cases and 116 (10.1%) of the 1149 controls were excluded from the GoldenGate assay due to low DNA amounts available at the time of analysis.
A SNPplexTM (Applied Biosystems) assay was used to genotype eight SNPs chosen to cover most of the common variation in XRCC2. SNP selection was based on tagSNP analysis (21) (r2 ≥ 0.8, minor allele frequency ≥0.05) of 97 individuals of European descent from the SNP500Cancer reference population. Because of low availability of DNA at the time of analyses or poor performance on the iPLEXTM assay, data were obtained on a subset of 946 cases (eight with buccal DNA samples) and 912 controls (32 with buccal DNA samples). Exclusions due to missing smoking information, non-transitional urothelial tumors and non-Caucasian descent resulted in 918 cases and 904 controls for the analyses.
Table I lists all SNPs determined using TaqMan, GoldenGate or iPLEX assays. TaqMan and iPLEX assays were performed at the Core Genotyping Facility of the Division of Cancer Epidemiology and Genetics, National Cancer Institute, and description and methods for assays can be found at http://snp500cancer.nci.nih.gov (22). All assays were performed using randomly sorted DNA samples from cases and controls, including blinded duplicate samples for quality control. Duplicate quality control DNA samples (up to 93 pairs) displayed >98% concordance except for XRCC2 promoter SNP rs3218374 (96%). All genotypes studied were in Hardy–Weinberg equilibrium in the control population. Pairwise linkage disequilibrium (LD) between SNPs was estimated based on D' and r2 values using Haploview (http://www.broad.mit.edu/mpg/haploview/index.php) (23).


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  		Table I. DSBR genes and SNPs evaluated in the Spanish Bladder Cancer Study

 
Statistical analysis
For each individual SNP, we estimated odds ratios (ORs) and 95% confidence intervals (95% CIs) using logistic regression models adjusting for gender, age at diagnosis in 5 year categories, region and smoking status (never, occasional, former and current). A global test for the association of the DSBR pathway genetic variants analyzed and bladder cancer risk was performed using a likelihood ratio test (LRT). The LRT compared a model with a single term for each genotype and covariates with a model with covariates only (degrees of freedom = number of SNPs). Gene–gene interactions, including GSTM1 and NAT2, and interactions of SNPs with age, gender and smoking habit were evaluated by introducing interaction terms in logistic regression models.

Haplotype frequencies for genes with more than one SNP were estimated using HaploStats (version 1.2.1; http://mayoresearch.mayo.edu/mayo/research/biostat/schaid.cfm). We evaluated the robustness of our results using the false discovery rate (FDR). FDR is the expected ratio of erroneous rejections of the null hypothesis to the total number of rejected hypothesis among all the SNPs analyzed in the report. Phylogenetic trees (neighbor-joining (24), nucleotide P distance) were constructed using MEGA 3.1 (25) (http://www.megasoftware.net/) to assess nucleotide similarity of different haplotypes.

The FDR method was applied to the P-value for trend applied only to results from 29 SNPs with r2 ≤ 0.90 evaluated in this report. Rather than using an arbitrary threshold FDR value, we report the values for the most significant associations to allow the reader to evaluate the robustness of our findings. The method by Benjamini et al. (26) was used to calculate FDR values using the package ‘multtest’ in the statistical program R (http://www.r-project.org/). Unless otherwise specified, statistical analyses were performed with STATA Version 9.1, Special Edition (STATA Corporation, College Station, TX).

Meta-analyses were performed to summarize our findings along with previously published studies for the association between the XRCC3 T241M and bladder cancer risk. Peer-reviewed studies with at least 100 cases published by October 2006 in English on this association were found using PubMed. Crude ORs and 95% CIs were obtained from published manuscripts. A random-effect model was used to estimate summary ORs and 95% CIs by weighing each study result by a factor within- and between-study variance (27). Homogeneity of study results was assessed by the Q test and publication bias by the test of Begg et al. (28) and Egger et al. (29).


   Results
Top
 Abstract
 Introduction
 Materials and methods
 Results
Discussion
 Funding
 Supplementary material
 References
 
The study population was of Caucasian descent, mostly males (87%) with a high proportion of cigarette smokers (40% of controls and 41% of cases were former smokers and 41% of cases and 23% of controls were current smokers) (17). Thirty-nine SNPs were analyzed in the current study corresponding to seven genes, three of which are involved in DNA break sensing, one in NHEJ and three in HR (Table I). To reduce redundancy, data from 10 of the 39 SNPs genotyped are not shown because they were highly correlated (r2 ≥ 0.90) with at least one other SNP (see supplementary Table 1, available at Carcinogenesis Online). A global test for the 29 non-redundant SNPs revealed a significant association between the DSBR variants evaluated and bladder cancer risk: global LRT (29 degrees of freedom) P = 0.01. Below we describe findings from individual SNP/gene analyses.

Individual SNP and haplotype analyses
Carriers of the R501S variant of ZNF350 (rs2278415) had a significantly lower risk of bladder cancer compared with individuals with the common homozygote genotype: OR (95% CI) for heterozygotes 0.80 (0.65–1.00) and for homozygotes 0.31 (0.11–0.83), P for trend = 0.01 (Table II, for selected results, and supplementary Table 1, available at Carcinogenesis Online, for complete results). We did not observe evidence for an interaction with age, gender or smoking status. Haplotype analysis of the three ZNF350 SNPs (5'–3' rs4988334, rs2278415 and rs2278414; D' ≥ 0.99, r2 ≥ 0.93) revealed two common haplotypes (>1%). Carriers of the three ZNF350 variants with the genotype CTT were at significantly decreased risk [0.78 (0.64–0.96)] as expected from our initial finding. Additionally, individuals with a second common haplotype present in 0.8% of controls with the genotype CAC were at significantly increased risk [1.97 (1.07–3.66)], and global P for haplotype analysis is 0.03 (data not shown). LD plots are shown in supplementary Figure 1 (available at Carcinogenesis Online).


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  		Table II. Association of eight selecteda polymorphisms in five DSBR genes on bladder cancer risk, adjusted for gender, age, region and smoking status (1150 cases and 1149 controls)

 
We observed a significant increase in risk for bladder cancer among carriers of a XRCC4 variant allele in intron 7 (rs1805377) compared with common homozygotes—OR (95% CI): 1.33 (1.08–1.64) per allele risk. An interaction with smoking was observed with the association being strongest for never smokers, weaker for former smokers and not present for current smokers: smoking-specific OR (95% CI) for never smokers 2.10 (1.32–3.35), former smokers 1.19 (0.86–1.65) and current smokers 1.02 (0.73–1.44); P for interaction LRT (2 degrees of freedom) is 0.05 (supplementary Table 2, available at Carcinogenesis Online). Moreover, an interaction with gender suggested a stronger association among females, [3.02 (1.54–5.95)] than males [1.20 (0.96–1.49)], P for interaction = 0.01 (data not shown). There was no evidence for an interaction with age. XRCC4 haplotype analysis based on four SNPs identified seven common (>1% frequent in controls) haplotypes in this population (Figure 1). Two haplotypes carried the two highly correlated SNPs in intron 7 that were individually associated with bladder cancer risk (rs2891980 and rs1805377; D' > 1.0, r2 > 1.0), and one of the two haplotypes carrying two additional variants (TACAA, 4.1% of controls) showed the strongest association with bladder cancer risk (Figure 1). A common haplotype (34.7% controls) that carried variants in the promoter and intron 4 (XRCC4 rs2075685, rs2662238; D' > 0.79, r2 > 0.59) was significantly associated with decreased risk [OR (95% CI) = 0.84 (0.72–0.98)] compared with the most common haplotype (Figure 1 and supplementary Table 1 and Figure 1, available at Carcinogenesis Online). These findings were consistent with individual SNP analyses, although associations were weaker and not statistically significant.


Figure 1
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  		Fig. 1. Phylogenetic tree of XRCC4 haplotypes and association with bladder cancer risk in the Spanish Bladder Cancer Study. **Polymorphic bases in 5'–3' order: XRCC4 rs2075685 rs2662238 rs2891980 rs3777015 and rs1805377. The rs numbers in bold and those in boxes denote SNPs that were associated with bladder cancer risk in individual genotype analysis. The referent group has the common variant for each individual SNP which is also the most common haplotype.

 
XRCC2 homozygote variants in three SNPs in the promoter (rs10234749, rs6464268, rs3218373) showed decreased risk compared with common homozygotes: OR (95% CI), respectively: 0.50 (0.31–0.81), 0.40 (0.19–0.84), 0.36 (0.13–0.96) (Table II). In addition, homozygotes for the R188H variant (rs3218536) were related to significant reduced risk [0.36 (0.13–1.00)]. Homozygote variants for all these SNPs were rare in the control population (≤3%) and showed no evidence for interaction with smoking, age or gender, except for XRCC2 rs6464268 LRT P value (2 degrees of freedom) is 0.05 (supplementary Table 2, available at Carcinogenesis Online). These variants were in LD (D' > 0.75) but were not highly correlated (r2 < 0.17). Haplotype analysis for this gene did not provide additional information above that of the individual associations seen (data not shown; LD plots shown in supplementary Figure 1, available at Carcinogenesis Online).

Robustness of our findings was evaluated using the FDR method, as described in the Materials and Methods. FDR values after accounting for the 29 SNPs tested with r2 > 0.90 for the SNPs with significant risk associations were 0.11 for ZNF350 rs2278415, 0.11 for XRCC4 rs1805377, 0.17 for XRCC2 rs6464268 and 0.61, 0.57 and 0.66 for XRCC2 rs10234749, rs3218373 and rs3218536, respectively.

Analysis of gene–gene pairwise interactions between the six SNPs significantly associated with bladder cancer risk or with the two established genetic susceptibility variants in GSTM1 and NAT2 (17) did not reveal any significant interactions (data not shown).

Meta-analysis
The XRCC3 non-synonymous SNP T241M (rs861539) has been related to bladder cancer risk in previous studies. To summarize these findings, we performed a meta-analysis using data from the six previous studies done in the USA, Italy and Sweden (2003 cases and 2140 controls) (30–35). The summary relative risk estimate for homozygous variant genotype versus the homozygous common genotype including our data was 1.17 (1.00–1.36), P = 0.05 (3086 cases and 3150 controls), suggesting a small increase in risk (Figure 2). We found no statistical evidence for publication bias or study heterogeneity, Q test P = 0.55.


Figure 2
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  		Fig. 2. Meta-analysis of XRCC3 rs861539 Ex8-53C>T, Thr241Met. Note: Common homozygote CC (Thr/Thr) is referent group. Studies are weighted and ranked according to the inverse of the variance of the log OR estimate. Q test for heterogeneity for CT P=0.07 and TT P=0.55.

 

   Discussion
Top
 Abstract
 Introduction
 Materials and methods
 Results
 Discussion
Funding
 Supplementary material
 References
 
Evaluation of 39 variants in seven DSBR pathway genes suggested that the genetic variants evaluated could contribute to bladder cancer risk (global LRT P = 0.01). Individual SNP analyses showed significant associations with risk in three genes with different roles in the DSBR pathway: ZNF350, XRCC4 and XRCC2.

ZNF350, which regulates the expression of DNA damage-responsive genes (9), showed an association with reduced bladder cancer risk. The functional significance of this association is not known, although other polymorphisms have been associated with cancer risk (36). The NBS1 gene is critical in DNA damage sensing and germ line mutations have been shown to predispose individuals to cancer (5,8). The NBS1 E185Q SNP (rs1805794) has been evaluated in relation to bladder cancer risk in two prior studies in the USA and Sweden (32,35), suggesting an association between the variant allele and increased risk. We found no significant association with this SNP in our study population; however, our data were consistent with a small increase in bladder cancer risk. Risk related to DNA break sensing genes is biologically plausible given recent reports that DNA damage responses are altered in bladder tumors (15).

The XRCC4 gene is necessary for DNA ligation in NHEJ (37,38), and consistent with a previous study of 696 cases and 629 controls in the USA (35), XRCC4 (rs1805377) was related with increased risk for bladder cancer. This SNP may have functional significance since the nucleotide change from G to A potentially abolishes an acceptor splice site at exon 8 (39). However, findings need to be replicated in additional study populations, particularly since homozygote variants were rare in the control populations [1% in our study and 2% in the study by Wu et al. (35)], and did not show significant associations with risk.

RAD51 is involved in HR, and in spite of having good coverage for common genetic variation for RAD51 (almost 90% of all genetic variants according to HapMap), we found no significant association with bladder cancer risk for any individual SNPs or haplotypes. In contrast, comprehensive evaluation of XRCC2 common genetic variation revealed reduced risk associations among homozygote variants for four rare SNPs, three of which were in the promoter region and the other a non-synonymous SNP R188H (rs3218536). Two previous studies assessed the association of the XRCC2 R188H with bladder cancer with conflicting results (31,35). Our study showed a significant protective association among homozygote variants with bladder cancer risk that was consistent with an Italian study of 317 cases and 317 controls (31). The other US study of 696 cases and 629 controls showed a protective association among heterozygotes but an increased risk among homozygotes (35); thus, further studies are needed to clarify this potential association.

The XRCC3 T241M (rs861539) variant has been shown to be functionally defective in suppressing duplication of the genome, which is thought to be important for maintaining genomic stability (40). Six previous studies in the USA, Italy and Sweden (30–35) including a total of 3112 cases and 3149 controls have evaluated its relationship with bladder cancer risk. The summary OR (95% CI) suggests a weak, though statistically significant, increase in risk with an OR of 1.17 (1.00–1.36). Even larger sample sizes are required to confirm or refute this potential recessive association.

FDR calculations indicated that the observed associations with bladder cancer risk for ZNF350 rs2278415, XRCC4 rs1805377 and XRCC2 rs6464268 are robust, and thus, follow-up in independent study populations is warranted. On the other hand, the associations between reduced risk of bladder cancer risk and the rare homozygous variant genotypes for three additional SNPs in the XRCC2 gene (rs10234749, rs3218373 and rs3218536) were not robust, and thus, findings need to be interpreted with caution. Strengths of our study included a large sample size with high participation rates and a relatively comprehensive analysis of genetic variation in DSBR. Except for XRCC2 and RAD51, we chose to evaluate SNPs prioritized by variants with potential functional significance since data were not yet available to ID-tagged SNPs throughout. Therefore, additional variation in DSBR not captured by the analyzed SNPs could or could not be associated with bladder cancer risk.

In conclusion, our results provide evidence for an association between genetic variants in the DSBR pathway and bladder cancer risk, which is biologically plausible given that germ line mutations in this pathway have been associated with cancer predisposition, and evidence that these mechanisms are important for maintaining genomic stability. Future studies are needed to confirm these findings.


   Funding
Top
 Abstract
 Introduction
 Materials and methods
 Results
 Discussion
 Funding
Supplementary material
 References
 
Intramural Research Program of the National Institutes of Health, National Cancer Institute, Division of Cancer Epidemiology and Genetics; FIS/Spain (00/0745, G03/174, G03/160, C03/09, C03/10).


   Supplementary material
Top
 Abstract
 Introduction
 Materials and methods
 Results
 Discussion
 Funding
 Supplementary material
References
 
Supplementary Table 1, Table 2 and Figure 1 can be found at http://carcin.oxfordjournals.org/.


   Acknowledgments
 
We thank Robert C.Saal from Westat, Rockville, MD, and Leslie Carroll and Jane Wang from IMS, Silver Spring, MD, for their support in study and data management; Doug Richesson from DCEG, NCI, for his support in data analysis, Dr Maria Sala from IMIM, Barcelona, Spain, for her work in data collection; Francisco Fernandez for his work on data management, Dr Montserrat Torà for her work in the coordination of sample collection and blood processing; and physicians, nurses, interviewers and study participants for their efforts during field work. Lastly, Jonine Figueroa would like to thank the NCI Division of Cancer Prevention, Cancer Prevention Fellowship Program for their support.

Participating Study Centers in Spain

Institut Municipal d'Investigació Mèdica, Universitat Pompeu Fabra, Barcelona—Coordinating Center (M.Kogevinas, N.Malats, F.X.Real, F.Fernandez, M.Sala, G.Castaño, M.Torà, D.Puente, C.Villanueva, C.Murta, J.Fortuny, E.López, S.Hernández, R.Jaramillo); Hospital del Mar, Universitat Autònoma de Barcelona, Barcelona (J.Lloreta, S.Serrano, L.Ferrer, A.Gelabert, J.Carles, O.Bielsa, K.Villadiego); Hospital Germans Tries i Pujol, Badalona, Barcelona (L.Cecchini, J.M.Saladié, L.Ibarz); Hospital de Sant Boi, Sant Boi, Barcelona (M.Céspedes); Centre Hospitalari Parc Taulí, Sabadell, Barcelona (C.Serra, D.García, J.Pujadas, R.Hernando, A.Cabezuelo, C.Abad, A.Prera, J.Prat); ALTHAIA, Manresa, Barcelona (M.Domènech, J.Badal, J.Malet); Hospital Universitario, La Laguna, Tenerife (R.García-Closas, J.Rodríguez de Vera, A.I.Martín); Hospital La Candelaria, Santa Cruz, Tenerife (J.Taño, F.Cáceres); Hospital General Universitario de Elche, Universidad Miguel Hernández, Elche, Alicante (A.Carrato, F.García-López, M.Ull, A.Teruel, E.Andrada, A.Bustos, A.Castillejo, J.L.Soto); Universidad de Oviedo, Oviedo, Asturias (A.Tardón); Hospital San Agustín, Avilés, Asturias (J.L.Guate, J.M.Lanzas, J.Velasco); Hospital Central Covadonga, Oviedo, Asturias (J.M.Fernández, J.J.Rodríguez, A.Herrero); Hospital Central General, Oviedo, Asturias (R.Abascal, C.Manzano); Hospital de Cabueñes, Gijón, Asturias (M.Rivas, M.Arguelles); Hospital de Jove, Gijón, Asturias (M.Díaz, J.Sánchez, O.González); Hospital de Cruz Roja, Gijón, Asturias (A.Mateos, V.Frade); Hospital Alvarez-Buylla, Mieres, Asturias (P.Muntañola, C.Pravia); Hospital Jarrio, Coaña, Asturias (A.M.Huescar, F.Huergo); Hospital Carmen y Severo Ochoa, Cangas, Asturias (J.Mosquera).

Conflict of Interest Statement: None declared.


   References
Top
 Abstract
 Introduction
 Materials and methods
 Results
 Discussion
 Funding
 Supplementary material
 References
 

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Received February 9, 2007; revised April 18, 2007; accepted May 25, 2007.


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L. M. Dong, J. D. Potter, E. White, C. M. Ulrich, L. R. Cardon, and U. Peters
Genetic Susceptibility to Cancer: The Role of Polymorphisms in Candidate Genes
JAMA, May 28, 2008; 299(20): 2423 - 2436.
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