[CANCER RESEARCH 59, 25572561, June 1, 1999]

Advances in Brief

XRCC1 Polymorphisms: Effects on Aflatoxin B1-DNA Adducts and Glycophorin A

Variant Frequency
Ruth M. Lunn, Ronald G. Langlois, Ling Ling Hsieh, Claudia L. Thompson, and Douglas A. Bell1
Laboratory of Computational Biology and Risk Assessment [R. M. L., D. A. B.] and Division of Extramural Research and Training [C. L. T.], National Institute of Environmental
Health Sciences, Research Triangle Park, North Carolina 27709; Lawrence Livermore National Laboratory, Livermore, California 94550 [R. G. L.]; and Department of Public
Health, Chang Gung College, Kwei-San, Tao-Yuan, Taiwan, Republic of China 10018 [L. L. H.]

Abstract repair gaps left during BER (9). PARP detects DNA strand breaks
induced by ionizing radiation and is believed to participate in BER
Hereditary genetic defects in DNA repair lead to increased risk of (10). XRCC1 negatively regulates PARP by binding to it via the
cancer. Polymorphisms in several DNA repair genes have been identified;
XRCC1 central domain (amino acids 301 402; Ref. 11). This central
however, the impact on repair phenotype has not been elucidated. We
explored the relationship between polymorphisms in the DNA repair
region also includes a BRCT domain and shares homology to the
enzyme, XRCC1 (codons 194, 280, and 399), and genotoxic end points yeast rad4/cut5 DNA repair gene (11, 12). Functional importance of
measured in two populations: (a) placental aflatoxin B1 DNA (AFB1-DNA) this region is also suggested by the determination that the DNA
adducts in a group of Taiwanese maternity subjects (n 120); and (b) repair-deficient EM-11 cell line contains a cysteine-to-tyrosine muta-
somatic glycophorin A (GPA) variants in erythrocytes from a group of tion at codon 390 (8).
North Carolina smokers and nonsmokers (n 59). AFB1-DNA adducts Shen et al. (1) identified three coding polymorphisms in the XRCC1
were measured by ELISA, and erythrocyte GPA variant frequency (NN gene at codons 194 (Arg to Trp), 280 (Arg to His) and 399 (Arg to
and N) was assessed in MN heterozygotes with a flow cytometric assay. Gln). These polymorphisms code for nonconservative amino acid
XRCC1 genotypes were identified by PCR-RFLPs. The XRCC1 399Gln changes (including the Arg399Gln change in the PARP binding
allele was significantly associated with higher levels of both AFB1-DNA
domain), which suggests potential functional relevance, but their
adducts and GPA NN mutations. Individuals with the 399Gln allele were
at risk for detectable adducts (odds ratio, 2.4; 95% confidence interval,
impact on phenotype is unknown. We tested whether XRCC1 poly-
1.15.4; P 0.03). GPA NN variant frequency was significantly higher in morphisms were associated with higher levels of genotoxic damage
399Gln homozygotes (19.6 10-6) than in Gln/Arg heterozygotes and found that the 399 Gln allele was significantly associated with
(11.4 10-6; P < 0.05) or Arg/Arg homozygotes (10.1 10-6; P 0.01). higher levels of AFB1-DNA adducts and GPA somatic mutations.
No significant effects were observed for other XRCC1 polymorphisms.
These results suggest that the Arg399Gln amino acid change may alter the Materials and Methods
phenotype of the XRCC1 protein, resulting in deficient DNA repair.
Subjects. AFB1-DNA adducts and XRCC1 genotypes were assessed in 120
Introduction placental DNA samples obtained from uncomplicated pregnancies at Taipei
Chang Gung Memorial Hospital as described by Hsieh and Hsieh (13). Sixty
Hereditary genetic defects in DNA repair lead to a marked in- placentas were collected during high (summer) and low (winter) exposure
creased risk of developing cancer. Although DNA repair deficiency season. GPA VF was measured in erythrocyte samples obtained from 49
often arises from mutations in genes that result in a loss of the DNA smokers and 10 nonsmokers (17 blacks and 42 whites) heterozygous for the
repair protein, DNA polymorphisms may alter the structure of the GPA antigen. The subjects were part of a community-based sample comprised
DNA repair enzyme and modulate cancer susceptibility. Mutations of 294 healthy unrelated blacks and whites from Durham and Chapel Hill,
North Carolina. This sample population has been used in other genotyping and
and polymorphisms have been identified in many of the genes coding
2 exposure studies (14, 15). XRCC1 genotypes were determined using genomic
for DNA repair enzymes such as XRCC1 (13). XRCC1 was identi- DNA isolated from lymphocytes. One hundred and ten additional whites and
fied by its ability to restore DNA repair activity in the Chinese 81 additional blacks from the same community sample were included in the
hamster ovary cell lines, EM-9 and EM-11, which are hypersensitive genotyping studies to compare XRCC1 allele frequency in different ethnic
to ionizing radiation and alkylating agents (4 6). These cells have groups.
increased spontaneous and mutagen-induced sister chromatid ex- Detection of AFB1-DNA Adducts. AFB1-DNA adduct levels were meas-
change and have defects in rejoining single-strand breaks after expo- ured by competitive ELISA using monoclonal antibody 6A10 and 50 g of
sure to X-ray (6, 7). Both EM-9 and EM-11 cells contain a mutated DNA as described previously (16). The percent of inhibition was calculated by
XRCC1 gene and lack XRCC1 protein (8). comparison with the nonmodified heat-denatured calf thymus DNA control.
Ionizing radiation and alkylating agents cause DNA base damage DNA samples were quantified relative to an imidazole ring-opened AFB1-
DNA standard, which has a modification level of 4 adducts/105 nucleotide.
and strand breaks that elicit BER. The XRCC1 protein complexes
Values below 20% inhibition, corresponding to 0.5 mol/mol DNA, were
with DNA ligase III via a BRCT domain in its COOH terminus and considered not detectable. Each sample was measured in triplicate on three
with DNA polymerase via the XRCC1 NH2 terminus domain to different assay dates and had a variability of less than 10%.
Measurement of GPA Variants in Erythrocytes. Blood samples were
Received 2/4/99; accepted 4/16/99. typed for the M and N antigens using commercial sera (Ortho Diagnostics,
The costs of publication of this article were defrayed in part by the payment of page Raritan, NJ) to determine MN heterozygous individuals. Fifty-nine individuals
charges. This article must therefore be hereby marked advertisement in accordance with
were identified and assessed for variants (N and NN) using the BR6 version
18 U.S.C. Section 1734 solely to indicate this fact.
1
To whom requests for reprints should be addressed, at LCBRA, National Institute of of the GPA assay as described previously (17, 18). A total of 5 106
Environmental Health Sciences, MD C3 03, P. O. Box 12233, Research Triangle Park, erythrocytes were analyzed for each sample.
NC 27709. E-mail: [email protected]. XRCC1 Genotyping. XRCC1 genotypes were detected using a PCR-RFLP
2
The abbreviations used are: XRCC1, X-ray repair cross-complementing 1; BER, base
technique. A multiplex PCR was used to amplify 491 bp and 615 bp of DNA
excision repair; BRCT, BRCA1 COOH terminus; PARP, poly (ADP-ribose) polymerase;
AFB1, aflatoxin B1; GPA, glycophorin A; OR, odds ratio; CI, confidence interval; VF, fragments containing the codon 194 and 399 polymorphisms, respectively.
variant frequency; LS, least square(s). Primers were: (a) 26106F gcc ccg tcc cag gta and 26577R agc ccc aag acc ctt
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 XRCC1 POLYMORPHISMS, AFLATOXIN ADDUCTS, AND GPA MUTATIONS

Statistical Analysis. Allele frequencies were estimated, and differences

between the various ethnic groups (blacks, whites, and Taiwanese) were tested
by pair-wise comparisons of 2 2 2 contingency table analysis. The asso-
ciation between XRCC1 alleles and AFB1-DNA adducts was evaluated by
traditional 2 k-table analysis (OR and 95% CI) and by a logistic regression
model controlling for season (adjusted OR and 95% CI). We evaluated the
effect of genotype on GPA variants by estimating the LS mean GPA VF (NN
and N) for each genotype, using an analysis of covariance model that
adjusted for age and smoking status.

Results and Discussion

Estimated genotype and allele frequencies for XRCC1 polymor-

phisms in black, white, and Taiwanese population samples are given
in Table 1. All of the distributions were in Hardy-Weinberg equilib-
rium. The 194Trp and 280His alleles were at a low frequency in
blacks [0.05 (194Trp) and 0.02 (280His)] and whites [0.06 (194Trp)
and 0.03 (280His)] but were significantly more prevalent in Taiwan-
ese [0.27 (194Trp) and 0.11 (280His) P 0.001]. The 399Gln-allele
frequency was significantly different among all of the three popula-
tions with the Gln allele occurring the highest in whites (0.37),
intermediate in Taiwanese (0.26) and lowest in blacks (0.17; Table 1).
Shen et al. (1) estimated variant allele frequency of 0.25 (194Trp),
0.08 (280His) and 0.25 (399Gln) by sequencing the XRCC1 gene from
12 unidentified individuals. These frequencies are most similar to that
observed in the Taiwanese population (Table 1).
To investigate whether the XRCC1 polymorphisms were associated
with differences in DNA repair, which might be reflected in levels of
genotoxic damage, we compared XRCC1 genotypes with levels of
AFB1-DNA adducts (Tables 2 and 3) and GPA somatic mutations
(Table 4). AFB1-DNA adducts in placental samples obtained from
Fig. 1. A, 3% Metaphor gel on MspI digestion of the multiplex XRCC1 PCR products 120 healthy Taiwanese women were measured by competitive ELISA,
containing codons 194 and 399. Codon 194 and 399 genotypes are indicated above and and the data were previously reported (13). The ELISA assay detects
below the Lanes, respectively. Uncut, the two undigested products (491-bp and 615-bp,
codons 194 and 399, respectively). Representative wild-type homozygotes (Arg/Arg and AFB1-adduct levels greater than 0.5 mol/mol DNA. Individuals
Arg/Arg, 194 and 399, respectively), heterozygotes (Arg/Trp and Arg/Gln, 194 and 399 with genotypes containing the 399Gln allele were more likely to have
respectively), and variant homozygotes (Trp/Trp and Gln/Gln, 194 and 399, respectively) detectable AFB1-DNA adducts (OR, 2.4; 95% CI, 1.15.4; P 0.03;
are shown for each polymorphism. M, 100-bp molecular weight standard (Promega). B,
2% NuSieve gel on RspI digestion of XRCC1 PCR product containing codon 280. Uncut,
depicts the 861-bp PCR product. Wild-type homozygotes (Arg/Arg) and heterozygotes
(Arg/His) are shown. Table 1 XRCC1 genotype and allele frequency among Taiwanese and North
Carolinian (NC) populations of whites (W) and blacks (B)
NC community
tca ct for codon 194; and (b) 27776F ttg tgc ttt ctc tgt gtc ca and 28371R tcc population
tcc agc ctt ttc tga ta for codon 399. A separate PCR using primers 27405F ttg Reported allele
acc ccc agt ggt gct aa and 28247R cgc tgg gac cac ctg tgt t were used to amplify Whites Blacks Taiwanese frequency
XRCC1 Polymorphisms (n 169) (n 98) (n 120) (n 12)a
the 861-bp DNA fragment containing the codon 280 polymorphism. PCR
conditions for both methods consisted of 50 ng of genomic DNA, 3 mM Exon 6,
Codon 194
MgCl2, 200 M each dNTPS, 0.5 units Taq (Promega, Madison, Arg/Arg 150 (89%) 89 (91%) 67 (56%)
WI) TaqStart Antibody (Sigma, St. Louis, MO), and either 0.6 M (codon Arg/Trp 18 (11%) 9 (9%) 42 (35%)
194) or 0.8 M (codon 280 and 399) each primer in 1 PCR buffer (Promega). Try/Trp 1 (0.5%) 0 (0%) 11 (8%)
PCR program was a 4-min denaturation step at 94C followed by 30 cycles of Allele frequency (Trp) 0.06b,c,d 0.05c,d 0.27d 0.25
30 s at 94C and 90 s at 68C. The Arg allele at codon 194 and the Arg allele Exon 9,
at codon 399 both create MspI sites. The multiplex 491-bp and 615-bp PCR Codon 280
products (codons 194 and 399, respectively) were digested at 37C for 2 h and Arg/Arg 159 (94%) 94 (96%) 95 (79%)
resolved on 3% Metaphor agarose gels (FMC Bioproducts, Rockland, ME; see Arg/His 10 (6%) 4 (4%) 23 (20%)
His/His 0 (0%) 0 (0%) 2 (2%)
Fig. 1A). A 174-bp fragment was present in all of the samples because of an c,d c,d
invariant MspI site (in the 491-bp fragment) that served as an internal control Allele frequency (His) 0.03 0.02 0.11d 0.08
for complete digestion. The Arg/Arg, Arg/Trp, and Trp/Trp genotypes for Exon 10
codon 194 resulted in 21-bp and 292-bp; 21-bp, 292-bp, and 313-bp; and Codon 399
Arg/Arg 65 (38%) 67 (69%) 63 (53%)
313-bp digestion products, respectively. The Gln allele (codon 399) was Arg/Gln 83 (49%) 27 (28%) 51 (43%)
distinguished from the Arg allele as an undigested fragment (615-bp) com- Gln/Gln 21 (13%) 3 (3%) 6 (4%)
pared with the 221- and 374-bp digested fragments of the Arg allele. RspI Allele frequency (Gln) 0.37d 0.17d 0.26d 0.25
digestion of the 861-bp PCR containing codon 280 was incubated separately at a
From Shen et al. (1).
37C for 2 h. The digestion fragments 60-bp, 221-bp, 580-bp, and 640-bp b
Allelic distribution for codon 194, 280, and 399 genotypes in Blacks (B), Whites
were separated on 2% 3:1 NuSieve agarose gels (FMC Bioproducts; see Fig. (W), and Taiwanese (T) sample populations were in Hardy Weinberg equilibrium.
c
1B). The Arg allele creates a RspI site at nucleotide 27466 and results in the Allele frequencies for W versus B were not significantly different: codon 194,
580-bp and 60-bp products that are not recognized by the allele but is P d Allele 0.7; codon 280, P 0.7.
frequency for the following were significantly different: codon 194, W versus
contained in the 640-bp fragment. The 221-bp fragment is a result of an T, B versus T, P 0.0001; codon 280, W versus T, B versus T, P 0.0005; codon 399,
invariant RspI site present in all of the samples. W versus B, P 0.0001; codon 399, W versus T, B versus T, P 0.05.
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 XRCC1 POLYMORPHISMS, AFLATOXIN ADDUCTS, AND GPA MUTATIONS

Table 2 The association between AFB1DNA adducts and XRCC1 polymorphisms

Nondetectable Detectable
adducts adducts
(n 51) (n 69) Crude ORa 95% CI; P Adjusted ORb 95% CI; P
Exon 6, codon 194
Arg/Arg 25 42 1.0c 1.0c
Arg/Trp 21 21 0.6 0.31.4; 0.27 0.5 0.21.2; 0.12
Trp/Trp 5 6 0.7 0.23.1; 0.86 0.7 0.22.6; 0.61
Arg/Trp Trp/Trp 26 27 0.6 0.31.3; 0.19 0.6 0.31.2; 0.14
Exon 9, codon 280
Arg/Arg 41 54 1.0c 1.0c
Arg/His 9 14 1.2 0.43.3; 0.91 1.1 0.42.9; 0.82
His/His 1 1 0.8 061.5; 0.60 0.7 012.9; 0.84
Arg/His His/His 10 15 1.1 0.43.1; 1.0 1.1 0.42.7; 0.87
Exon 10, codon 399
Arg/Arg 33 30 1.0c 1.0c
Arg/Gln 17 34 2.2 1.05.1; 0.07 2.7 1.26.1; 0.017
Gln/Gln 1 5 5.5d 0.6131; 0.21 6.4 0.760.8; 0.11
Arg/Gln Gln/Gln 18 39 2.4 1.15.4; 0.03 2.9 1.36.4; 0.008
a
Fisher exact OR.
b
OR adjusted for season, calculated by logistic regression.
c
Reference group.
d
Test for trend 2 5.1; P 0.024.

Table 2). Moreover, a gene-dosage effect was observed (test for trend, may help determine whether this possible difference is real or is due
X2 5.12; P 0.024). That is, individuals homozygous for the to chance. As reported by Hsieh and Hsieh (13), AFB1-DNA adducts
399Gln allele had a higher risk of having detectable AFB1-DNA were observed to be higher during the summer than the winter. The
adducts (OR, 5.5; 95% CI, 0.6 131; P 0.2) than heterozygous association between DNA damage and the 399Gln allele was similar
individuals (OR, 2.2; 95% CI, 1.15.1; P 0.07). However, the for AFB1-DNA adducts detected in samples collected in the summer
number of homozygous individuals was very small (n 6), limiting (OR, 2.7; (95% CI, 0.711.3; P 0.16) and winter (OR, 3.0; 95% CI,
the interpretation of this finding. No statistically significant associa- 1.0 10.2; P 0.04; data not shown). Adjusting for seasonal variation
tion was observed between the detection of AFB1-DNA adducts and did not substantially modify the ORs for the association of XRCC1
the 194Trp or 280His alleles; however, individuals carrying a 194Trp genotypes and AFB1-DNA adducts (Table 2).
allele were slightly more common in the nondetectable AFB1-DNA We examined the relationship of XRCC1 genotypes and different
adduct group (OR, 0.6; 95% CI, 0.31.3; P 0.19). A larger study levels of AFB1-DNA adducts (Table 3). Detectable AFB1-DNA ad-

Table 3 Codon 399 polymorphisms and AFB1-DNA adducts stratified by level of adducts
Codon 399 genotype
AFB1DNA
adduct level Arg/Arg Arg/Gln Gln/Gln ORa 95% CI; P ORb 95% CI; P
c d
Nondetectable 33 18 1.0 1.0d
Intermediatec 11 23 3.8 1.410.7; 0.004 5.2 1.914.3; 0.002
Highc 19 16 1.5 0.64.1; 0.38 1.5 0.92.4; 0.1
Detectable 30 39 2.4 1.15.4; 0.03 2.9 1.36.4; 0.008
(intermediate high)
a
Fisher exact OR.
b
OR adjusted for season, calculated by logistic regression.
c
Nondetectable, values are below the limit of detect for the assay; Intermediate, measurable values were less than or equal to the median (2.1 mol/mol DNA); High, values greater
than the median level of adducts.
d
Reference group.

Table 4 The association of GPA NN and N variants and XRCC1 genotypes

Mean GPA N VF (per 106 cells) Mean GPA NN VF (per 106 cells)

All subjectsa Smokersb Nonsmokersb All subjectsa Smokersb Nonsmokersb

c c c c c
Genotypes N LS mean SE N LS mean SE N LS mean SE N LS mean SE N LS mean SE N LS mean SEc
Exon 6, Codon 194
Arg/Arg 55 13.6 0.9 45 12.9 0.9 10 16.9 2.9 55 11.6 1.0 45 11.1 1.1 10 13.5 2.7
Arg/Trp 4 13.1 3.5 4 12.9 3.2 0 4 10.4 4.0 4 11.0 3.9 0
Trp/Trp 0 0 0 0 0
Exon 9, Codon 290
Arg/Arg 56 13.6 0.9 47 12.9 0.9 9 17.1 3.2 56 11.5 1.0 47 11.1 1.1 9 13.3 3.0
Arg/His 3 13 3.9 2 13.3 4.3 1 15.0 9.7 3 11.8 4.5 2 11.1 5.3 1 15.2 9.0
His/His 0 0 0 0 0
Exon 10, Codon 399
Arg/Arg 31 13.3 1.2 26 12.6 1.2 5 18.1 4.8 31 10.1d 1.3 26 10.0d 1.4 5 12.1 4.4
Arg/Gln 22 13.9 1.4 19 13.3 1.4 3 16.1 6.0 22 11.4e 1.5 19 10.6e 1.6 3 15.5 5.5
Gln/Gln 6 13.5 2.8 4 12.4 3.1 2 14.9 7.9 6 19.6d,e 3.0 4 21.0d,e 3.5 2 14.1 7.2
a
Adjusted for smoking and age.
b
For smokers or nonsmokers adjusted for age.
c
Standard error of the mean.
d
P 0.01, Gln/Gln versus Arg/Arg.
e
P 0.05, Gln/Gln versus Arg/Gln.
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 XRCC1 POLYMORPHISMS, AFLATOXIN ADDUCTS, AND GPA MUTATIONS

ducts levels ranged from 0.6 to 6.3 mol/mol DNA with a median of tions in GPA VF, thus implicating it as a marker of exposure, damage,
2.1 mol/mol DNA. We classified individuals as nondetectable and cancer risk (22). N and NN variants arise from independent
( 0.5 mol/mol DNA), intermediate ( 0.5 and 2.1 mol/mol molecular mechanisms (22). Gene inactivation mechanisms such as
DNA), and high ( 2.1 mol/mol DNA). Individuals with genotypes point mutations, deletions, and chromosome loss are likely to result in
containing the 399Gln allele were more likely to have an intermediate N variants, whereas mitotic recombination, gene conversion, and
level of adducts (OR, 3.8; 95% CI, 1.4 10.7; P 0.004) than a high chromosome missegregation are more important in generating NN
level of adducts (OR, 1.5; 95% CI, 0.6 6.4; P 0.38). No significant variants (23). The relationship of 399Gln allele to NN but not N
association occurred between XRCC1 polymorphisms at codon 194 variants may result from a greater role of the central domain of
and 280 and adducts (data not shown). XRCC1 in recombination repair than repair of lesions that cause gene
AFB1 mediates its carcinogenicity mainly through the formation of inactivation. Both PARP and XRCC1 participate in DNA strand-break
AFB1-guanine adducts. These highly unstable adducts can either form rejoining and homologous recombination (7, 24 26). Cells without
more stable ring-opened structures or undergo spontaneous depurina- the XRCC1 gene and mice lacking the PARP gene exhibit high levels
tion, resulting in apurinic sites and eliciting BER (19 21). Adminis- of sister chromatid exchange, which suggests increased recombination
tration of AFB1 in rats causes single-strand breaks and increases activity (7, 26). PARP inhibitors cause an increase in recombination
PARP, DNA ligase, and DNA polymerase enzyme activity (20). frequency and genomic instability (27), and XRCC1 expression is
These enzymes interact with XRCC1 during BER, suggesting that elevated during male meiosis in the mouse, which implies a role in
XRCC1 may be important in the repair of AFB1-DNA adducts. Thus, meiotic recombination (28).
the association of AFB1-DNA adducts and the 399Gln allele is bio- This is the first report to investigate associations between pheno-
logically plausible. Moreover, codon 399 is located in the PARP- type (measures of genotoxic damage) and three missense polymor-
binding region of the XRCC1 gene. The specific functional effect of phisms194(Arg to Trp), 280 (Arg to His), and 399 (Arg to Gln)in
the Arg399Gln change on XRCC1 binding with PARP remains to be the XRCC1 gene. We find evidence to suggest that the XRCC1 399Gln
explored. allele is associated with increased levels of DNA damage that may be
due to reduced DNA repair function. Individuals with the Gln allele
Although the 399Gln allele of XRCC1 was related to the detection
were more likely to have higher levels of AFB1-DNA adducts and
of AFB1-DNA adducts, the effect seems to be greatest at lower adduct
GPA NN somatic variants. BER is important in the repair of AFB1-
levels (Table 3). Possibly, in tissues with higher levels of AFB1-DNA
DNA adducts, whereas errors in recombination may generate GPA
adducts, the BER pathway may become saturated, which would tend
NN variants. XRCC1 is implicated in both of the repair processes.
to reduce differences between functional and less functional alleles. A
Moreover, the Arg399Gln polymorphism occurs in a region of the
similar phenomena has been observed in rodent exposure studies in
XRCC1 gene that contains biologically important domains (PARP
which very high levels of AFB1-induced DNA damage resulted in a
binding and BRCT) and has homology with another DNA repair-
decline of PARP activity. (20).
related gene (yeast rad4/cut5 gene). Future studies need to character-
Differences in AFB1-DNA adducts also reflect differences in ex-
ize the role of the XRCC1 399Gln allele in functional DNA repair
posure. External exposure information was not available for the sub-
assays and to test to see whether it affects the levels of other biomar-
jects so that the direct effects of adduct repair cannot be assessed. In kers of DNA damage.
vitro or in vivo studies using a fixed AFB1 exposure should provide
insight into the effect of genotype on the repair of AFB1-DNA
Acknowledgments
adducts.
We also examined the relationship of XRCC1 genotypes and so- We thank Drs. Harvey Mohrenweiser and Richard Shen for sharing infor-
matic mutations characterized by the GPA assay (Table 4). The GPA mation about XRCC1 polymorphisms. We also thank the following: Richard
assay detects two types of allele loss variants N (allele loss) and NN Morris and Dr. Xugang Guo for assistance with statistical analysis; Kathleen
(allele loss and duplication) present in erythrocytes. We measured the Bones for technical support; and Gary Pittman and Dr. William Kaufmann for
critical review of the manuscript.
VF of both N and NN mutations in smokers and nonsmokers who
were heterozygous for GPA. The mean VF (N and NN) was esti-
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XRCC1 Polymorphisms: Effects on Aflatoxin B1-DNA Adducts
and Glycophorin A Variant Frequency
Ruth M. Lunn, Ronald G. Langlois, Ling Ling Hsieh, et al.

Cancer Res 1999;59:2557-2561.

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