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RESEARCH ARTICLE
Association between RECQL5 genetic polymorphismsand susceptibility to breast cancer
Yu-Jun He & Zuo-Yi Qiao & Bo Gao & Xiao-Hua Zhang &
Ya-Yuan Wen
Received: 15 July 2014 /Accepted: 20 August 2014# International Society of Oncology and BioMarkers (ISOBM) 2014
Abstract Previous studies indicated that the RECQL5 genepolymorphism was associated with human cancers. However,the association of RECQL5 gene polymorphism with breastcancer remains unclear. In the present study, we investigatedthe association between polymorphisms of the RECQL geneand breast cancer in a Chinese population. We selected fourpolymorphisms of the RECQL5 gene (rs820186, rs820196,rs820200, and rs4789223) for the present study. Thegenotyping was performed using the TaqMan method in510 patients with breast cancer and 510 age- and sex-matched non-cancer controls. We found that rs820196 andrs828200 polymorphisms of RECQL5 were associated withbreast cancer. For rs820196, the CC genotype (16.7 vs9.4 %, P<0.001) and C allele (42.5 vs 34.3 %, P<0.001)were common in the breast cancer patients than in thecontrol subjects, respectively. For rs828200, the GG genotype(23.7 vs 18.0 %, P<0.001) and G allele (52.7 vs 43.8 %,P<0.001) were common in the breast cancer patients than inthe control subjects, respectively. Haplotype analysis showed
that C-G (odds ratio (OR)=2.247, 95 % confidence interval(CI) 1.854∼2.722; P<0.001) was associated with increasedrisk for breast cancer. However, the C-T (OR=0.175, 95 % CI0.110∼0.278; P<0.001) and T-G (OR=0.544; 95 % CI0.428∼0.692; P<0.001) were associated with decreased riskfor breast cancer, respectively. The present study indicatedthat the RECQL5 genetic polymorphism and haplotypes wereassociated with breast cancer in a Chinese population.
Keywords RECQL5 . Breast cancer . Polymorphism .
Haplotype
Introduction
Breast cancer is now the most frequently diagnosed cancerand the leading global cause of cancer death in women,accounting for 23 % of cancer diagnoses (1.38 million women)and 14 % of cancer deaths (458,000 women) each year [1].Although breast cancer has a markedly higher incidence indeveloped countries, half of new breast cancer diagnoses andan estimated 60 % of breast cancer deaths are now thought tooccur in the developing world [2]. The incidence and mortalityrate in developing countries are even higher because of limitedmedical infrastructure and awareness [3, 4]. China, one of thedeveloping countries, has a high incidence of breast cancer inthe world.
As with most human cancers, breast cancer is a complexdisease that maybe is a result of the interaction betweenenvironmental factors and genetics [5–9]. Previous studiesindicated that DNA damage, which, if not repaired ormisrepaired, may result in genomic instability, cancer trans-formation, or cell death [10, 11].
The RECQ family is a highly conserved group of DNAhelicases required for the maintenance of genome stability and
Y.<J. He : B. Gao :X.<H. Zhang :Y.<Y. Wen (*)Department of General Surgery, Daping Hospital and ResearchInstitute of Surgery, Third Military Medical University,Chongqing 400042, Chinae-mail: [email protected]
Y.<J. Hee-mail: [email protected]
B. Gaoe-mail: [email protected]
X.<H. Zhange-mail: [email protected]
Z.<Y. QiaoDepartment of Emergency, PLA 324 Hospital, Chongqing 400020,Chinae-mail: [email protected]
Tumor Biol.DOI 10.1007/s13277-014-2528-2
integrity. They have an important role in DNA replication,telomere maintenance, DNA damage signaling, and DNArepair pathways including mismatch repair, nucleotide exci-sion repair, and direct repair [12, 13]. Previous studies indi-cated that the mutation in RECQL family gene was associatedwith human cancer [12, 14, 15]. And RECQL5 haplotype wasreported to be associated with osteosarcoma [16] and laryn-geal cancer [17] previously. However, the relation betweenRECQL5 polymorphism and breast cancer remains unknown.In the present study, we investigated the relationship betweenRECQL5 polymorphisms and breast cancer in a Chinesepopulation.
Material and methods
Patients
The present study was approved by the Ethics Committee ofthe Third Military Medical University and each patient gavewritten consent. Blood samples were obtained from all thesubjects including 510 patients with breast cancer from May2007 to June 2014 and 510 cancer-free age-matched controls.The diagnosis of breast cancer was established by pathologicalexamination, mammography, core needle breast biopsy, andopen surgical breast biopsy. Epidemiological history such asage at onset of breast cancer, diet, socioeconomic status,occupation, reproductive history, family history, and
consanguinity was taken through personal interview withbreast cancer patients using specific proforma. All the breastcancer patients were recruited from the Third Military MedicalUniversity of China. The healthy controls were frequency-matched to the patients by age and were randomly selectedfrom the routine physical examination program in the samedistrict. All the healthy subjects had no documented history ofcancer or autoimmune diseases. It was ensured that the enrolledpatients and controls were of the Han ethnicity. The patientsranged in age from 35 to 76 years (54.3±7.9), and the controlsubjects ranged in age from 35 to 78 years (55.5±9.6).
Genotyping
According to the previous studies [16, 17], we selected fourtagging single nucleotide polymorphisms (SNPs) (rs820186,rs820196, rs820200, and rs4789223) for the present study.Genomic DNA was extracted from the peripheral blood leu-kocytes using a DNA extraction kit (Beijing Bioteke Co. Ltd).Genotyping was confirmed by the TaqMan method as de-scribed previously [16, 17].
Statistical analysis
We used the SPSS 17.0 for Windows (IL, CA, USA) for thestatistical analysis. We assessed the Hardy-Weinberg equilib-rium using the standard χ2 test or Fisher’s exact test. Thedistributions of genotype and allele frequencies between two
Table 1 Genotype distribution ofRECQL5 tag SNPs between caseand control subjects
SNPs Genotype and allele Breast cancer (n=510) Control (n=510) P value
rs820186 AA 15 (0.029) 13 (0.025) 0.752
AG 172 (0.337) 163 (0.320)
GG 323 (0.633) 334 (0.655)
A 202 (0.198) 189 (0.185) 0.464
G 818 (0.802) 831 (0.815)
rs820196 CC 85 (0.167) 48 (0.094) <0.001
CT 263 (0.516) 254 (0.498)
TT 162 (0.318) 208 (0.408)
C 433 (0.425) 350 (0.343) <0.001
T 587 (0.575) 670 (0.657)
rs820200 GG 121 (0.237) 92 (0.180) <0.001
GT 296 (0.580) 263 (0.516)
TT 93 (0.182) 155 (0.304)
G 538 (0.527) 447 (0.438) <0.001
T 482 (0.473) 573 (0.562)
rs4789223 AA 73 (0.143) 76 (0.149) 0.201
AG 227 (0.445) 249 (0.488)
GG 210 (0.412) 185 (0.363)
A 373 (0.366) 401 (0.393) 0.264
G 647 (0.634) 619 (0.607)
Tumor Biol.
groups were compared by χ2 tests. The genotype-specificrisks were estimated as odds ratios (ORs) and their 95 %confidence interval (CI). Linkage disequilibrium (LD) analy-ses and haplotype analyses were performed using the SHEsissoftware (http://analysis2.bio-x.cn/myAnalysis.php) [18, 19].Statistical significance was established at P<0.05.
Results
We did not find significant difference from the Hardy-Weinberg equilibrium values (data not shown) of these fourSNPs. As shown in Table 1, we found that rs820196 andrs828200 polymorphisms of RECQL5 were associated withbreast cancer. For rs820196, the CC genotype (16.7 vs 9.4 %,P<0.001) and C allele (42.5 vs 34.3 %, P<0.001) werecommon in the breast cancer patients than in the controlsubjects, respectively. For rs828200, the GG genotype (23.7vs 18.0 %, P<0.001) and G allele (52.7 vs 43.8 %, P<0.001)were common in the breast cancer patients than in thecontrol subjects, respectively. The genotype and the alleledistributions of other two SNPs (rs820186 and rs4789223)were not different between the breast cancer patients andthe control participants.
Figure 1 shows the patterns of LD in the RECQL5 gene,with their r2 values. All four SNPs are located in one haplo-type block. As shown in Table 2, haplotype analysis showedthat C-G (OR=2.247, 95 % CI 1.854∼2.722; P<0.001) wasassociated with increased risk for breast cancer. However, theC-T (OR=0.175, 95 % CI 0.110∼0.278; P<0.001) and T-G(OR=0.544; 95%CI 0.428∼0.692;P<0.001) were associatedwith decreased risk for breast cancer, respectively.
Discussion
In our study, we found that RECQL5 gene polymorphisms(rs820196 and rs828200) and haplotypes were significantlyassociated with breast cancer risk in a Chinese population.And this is the first study to reveal the association betweenRECQL5 genetic polymorphisms and susceptibility to breastcancer.
As described previously, RECQ helicases comprise a high-ly conserved family of DNA helicases that operate to maintaingenomic DNA stability in humans [20]. Defects in humanRECQ helicases are associated with premature aging, genomeinstability, and cancer predisposition [21]. RECQ helicasefamily members possess intrinsic DNA-dependent ATPaseand ATP-dependent DNA-unwinding activity, with variablepreferences for specific DNA structures. Recently, Pellatt et al.reported that RECQL5 polymorphism was associated withtelomere length and colon cancer [22].
Recent publications indicated that the genetic polymor-phisms of RECQL5 are associated with human cancer, suchas laryngeal cancer [17] and osteosarcoma [16]. Zhi et al. [16]explored the relation between osteosarcoma risk and RECQL5polymorphism and found that the CC genotype in rs820196(17.5 vs 8.3 %, P=0.005) and AA genotype in rs4789223(21.7 vs 14.2 %, P<0.001) were more frequent in the osteo-sarcoma group compared to the control group, respectively. Qiet al. [17] studied the association between laryngeal cancerrisk and RECQL5 polymorphisms and found that thers820196 polymorphism of RECQL5 was associated withlarynx cancer; the CC genotype (16.4 vs 9.3 %, P=0.013)and C allele (42 vs 34.2 %, P=0.006) were common in larynxcancer patients than in the control subjects, respectively. How-ever, rs4789223 was not found to be associated with larynxcancer in Qi et al.’s study.
Fig. 1 The patterns of linkage disequilibrium in the RECQL5 gene, withtheir r2 values
Table 2 Distribution ofhaplotypes Haplotype Breast cancer Control P value OR (95 % CI)
C-G 410.95 (0.403) 235.59 (0.231) <0.001 2.247 (1.854∼2.722)C-T 22.05 (0.022) 114.41 (0.112) <0.001 0.175 (0.110∼0.278)T-G 127.05 (0.125) 211.41 (0.207) <0.001 0.544 (0.428∼0.692)T-T 459.95 (0.451) 458.59 (0.450) 0.951 1.005 (0.844∼1.197)
Tumor Biol.
In our study, we genotyped four SNPs in RECQL5 inChinese participants and assessed the association betweenRECQL5 and breast cancer using a haplotype-based case-control analysis. The rs820196 and rs828200 significantlydiffered between breast cancer patients and control participants,indicating that the risk of breast cancer is increased in partici-pants with the C allele of rs820196 and G allele of rs828200.Morris and Kaplan found that for genes with multiple suscep-tibilities, analysis based on haplotypes has advantages overanalysis based on individual SNPs [23]. Consequently, in thepresent study, we successfully established haplotypes for theRECQL5 gene from the different combination of the two SNPs.The frequency of the C-T and T-G was associated with de-creased risk for breast cancer, respectively. However, the C-Gwas associated with increased risk for breast cancer.
In conclusion, the present results indicate that breast canceris associated with the RECQL5 gene polymorphisms. The C-G haplotype appears to be a useful genetic marker, and the C-T and T-G haplotypes might be protective factors of breastcancer in Chinese people.
Acknowledgments This work was supported by the Natural ScienceFoundation of Chongqing (CSTC, 2011BB5032).
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