research article lack of associations between xpc gene...

Research ArticleLack of Associations between XPC Gene Polymorphisms andNeuroblastoma Susceptibility in a Chinese Population

Jintao Zheng12 Ruizhong Zhang2 Jinhong Zhu3 Fenghua Wang2 Tianyou Yang2

Jing He2 and Huimin Xia12

1Department of Pediatric Surgery Foshan Maternity and Childrenrsquos Healthcare Hospital Affiliated to Southern Medical UniversityFoshan China2Department of Pediatric Surgery Guangzhou Institute of Pediatrics Guangzhou Women and Childrenrsquos Medical CenterGuangzhou Medical University Guangzhou Guangdong 510623 China3Molecular Epidemiology Laboratory and Department of Laboratory Medicine Harbin Medical University Cancer Hospital HarbinHeilongjiang 150040 China

Correspondence should be addressed to Jing He hejing198374gmailcom and Huimin Xia xia-huiminfoxmailcom

Received 11 July 2016 Accepted 27 September 2016

Academic Editor Hai-Feng Pan

Copyright copy 2016 Jintao Zheng et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Neuroblastoma is one of the most malignant solid tumors in infants and young children No more than 40 of neuroblastomapatients can survive for longer than five years after it has been diagnosed XPC protein is a pivotal factor that recognizes DNAdamage and starts up the nucleotide excision repair (NER) in mammalian cells This makes up the first group to defend against thecancer Previous studies have identified that XPC gene polymorphisms were associated with various types of cancer However theassociations betweenXPC gene polymorphisms and neuroblastoma risk have not yet been studiedWe investigated the associationsbetween three XPC gene polymorphisms (rs2228001 AgtC rs2228000 CgtT and rs2229090 GgtC) and neuroblastoma risk with 256neuroblastoma patients and 531 healthy controls in a Chinese Han population Odds ratios and 95 confidence intervals wereused to access the association between these three polymorphisms and neuroblastoma risk No significant association was detectedbetween these three polymorphisms and neuroblastoma risk in the overall analysis as well as in the stratification analysis Theseresults suggest that none of these three polymorphisms may be associated with the risk of neuroblastoma in the Chinese Hanpopulation

1 Introduction

Neuroblastoma originates in primitive neural crest cells ofthe adrenal medulla or sympathetic ganglia It is one of themost malignant solid tumors in infants and young childrenin particular accounting for 7minus10 of childhood tumorsThe tumor is found to primarily locate in the retroperitonealparts (approximately 60) and secondly in the mediastinumpelvis and cervical sympathetic ganglion The rest about12 associates with other malformations [1ndash3] The mor-bidity of neuroblastoma in the live births is about 77 casesper million in China [4] Despite the application of mul-timodality treatment including surgery chemotherapy andradiotherapy no more than 40 of neuroblastoma patients

could survive for longer than five years after diagnosis Untilnow the etiology of neuroblastoma remains largely unclear[5 6]

Genome-wide association study (GWAS) has been usedas a convenient and powerful approach for examining inher-ited genetic variations in relation to human complex diseasesuch as cancer susceptibility [7 8] In 2008 Maris et al[9] used the GWAS approach to prove that three singlenucleotide polymorphisms (SNPs) in CASC15 gene at chro-mosome 6p22 (rs6939340 rs4712653 and rs9295536) werehighly associated with neuroblastoma susceptibility in Euro-pean descents Since then lots of SNPs in specific chromo-somal regions have been identified by the GWAS approach

Hindawi Publishing CorporationBioMed Research InternationalVolume 2016 Article ID 2932049 6 pageshttpdxdoiorg10115520162932049

2 BioMed Research International

Several gene polymorphisms are significantly associated withthe risk of neuroblastoma such as LIN28B [10] HACE1 [10]BARD1 [11] LMO1 [12] HSD17B12 [13] DDX4 [13] IL31RA[13] and DUSP12 [13] which have been discovered by meansof GWAS Besides several genes such as FAS [14] FASL[14] NEFL [15] and TGFBR3L [16] have been discovered bycandidate gene approach

Nucleotide excision repair (NER) can preserve the hered-itary integrity and stability of genomic DNA through elim-inating DNA damage generated by exogenous agents (iemutagenic and carcinogenic substances) and photoproductscaused by sunlight exposure [17 18] In humans hereditarydefects of genomic DNA in NER are involved in several auto-somal recessive diseases Xeroderma pigmentosum (XP) ischaracterized by a strong predisposition to skin carcinomasHowever it was not until 1968 that Cleaver [19] identified thatXP could result from the defects in NER NER is a complexbiochemical process that involves hundreds of componentsin human cells including the seven XP-related genes (XPAXPB XPC XPD XPE XPF and XPG) XPC protein is animportant DNA damage recognition protein and an initiatorof the NER process that defends against cancer [20ndash25]

Some XPC gene SNPs have been identified to be impli-cated in melanoma [26] colorectal cancer [27 28] prostatecancer [29] hepatocellular cancer [30] lung cancer [31]and gastric cancer [32] However the association betweenXPC gene polymorphisms and neuroblastoma risk has notbeen studied In view of this we investigated the relation-ship between XPC gene polymorphisms (rs2228001 AgtCrs2228000 CgtT and rs2229090 GgtC) and neuroblastomasusceptibility in a Chinese Han population making use of atotal of 787 participants (256 cases and 531 controls)

2 Materials and Methods

21 Study Subjects A total of 256 newly diagnosed andhistopathologically confirmed neuroblastoma cases wererecruited from the Guangzhou Women and Childrenrsquos Med-ical Center between February 2010 and November 2015 [33ndash35] During the same period 531 healthy children were ran-domly selected as the age- and gender-matched controls afterreceiving routine physical examination [35ndash37] Both thecases and the controls were unassociated ethnic Chinese Hanindividuals The study was granted permission by the Insti-tutional Review Board of GuangzhouWomen and ChildrenrsquosMedical Center Demographic factors and medical historieswere gathered through the structured questionnairesWritteninformed consent was received from the guardians of eachchild participant Each participant donated 2mL of blood forgenomic DNA extraction

22 Polymorphism Analysis The selection criteria weredescribed previously in terms of the minor allele frequencyputative functional potentials and low linkage disequilib-rium [38] Based on the criteria three potentially functionalSNPs (rs2228001 AgtC rs2228000 CgtT and rs2229090 GgtC)were selected s2228001 AgtC (Lys939Gln) and rs2228000CgtT (Val499Arg) are nonsynonymous SNPs and they havebeen widely investigated in various types of cancer XPC

rs2229090 GgtC is 31015840UTR polymorphism within miRNAbinding site Genomic DNA was extracted from 2mL ofperipheral blood sample using the TIANamp Blood DNAKit (TianGen Biotech Co Ltd Beijing China) following themanufacturerrsquos instructions Qualified DNA samples werediluted to 10 ng120583L and loaded in 96-well plates Then thesethree SNPs were genotyped using Taqman real-time PCRmethod as described previously [39 40]

23 Statistical Analysis 1205942 test was used to evaluate the dif-ferences in the frequency distributions of the demographicsand genotypes between the neuroblastoma cases and thecontrols Hardy-Weinberg equilibrium (HWE) was tested inthe controls by the goodness-of-fit chi-squared test Oddsratios (ORs) and 95 confidence intervals (CIs) were usedto assess the correlations between the three polymorphismsand neuroblastoma susceptibility with the unconditionalmultivariate logistic regression analysis 119875 value lt 005 wasconsidered as statistically significant All statistical tests weretwo-sided and analyzed using SAS software (version 91 SASInstitute Cary NC)

3 Results

31 Frequency Distribution of Selected Characteristics Asdisplayed in Supplemental Table 1 in SupplementaryMaterialavailable online at httpdxdoiorg10115520162932049the average age of the cases was 3087 plusmn 2645 monthsand 2973 plusmn 2486 months for the controls No significantdifferences were observed in terms of age (119875 = 0239) andgender (119875 = 0333) between the case and the control groupsAccording to the INSS standard [3] 54 65 44 77 and 9patients developed clinical stages I II III and IV and 4sneuroblastoma respectively Among these cases 46 lesionsoccurred in adrenal gland 87 in retroperitoneal region and90 in mediastinum

32 XPC Gene Polymorphisms and Neuroblastoma Suscepti-bility In the current study 253 cases and 531 controls weresuccessfully genotypedThe genotype frequencies of the threepolymorphisms are shown in Table 1 The observed genotypefrequencies of the three SNPs were in accordance with HWEin the control subjects (119875 = 0948 for rs2228001 AgtCpolymorphism 119875 = 0988 for rs2228000 CgtT polymor-phism and 119875 = 0994 for rs2229090 GgtC polymorphism)There is no significant association between rs2228001 AgtCpolymorphism and neuroblastoma susceptibility Similarresults were found for rs2228000 CgtT and rs2229090 GgtCpolymorphisms

33 Stratification Analysis of XPC Gene Polymorphisms withNeuroblastoma Susceptibility Stratified analyses were con-ducted regarding age gender sites of origin and clinicalstages to assess the association of the three selected polymor-phisms with the risk of neuroblastoma (Table 2) Howeverno significant association was identified for any of the threepolymorphisms

BioMed Research International 3

Table1Genotypea

ndallelefre

quencies

ofthethree

selected

polymorph

ismsa

ndneurob

lasto

mas

usceptibilityin

aChinese

popu

lation

Genotype

Cases(119873=253)

Con

trols(119873=531)

119875a

Crud

eOR(95

CI)

119875Ad

juste

dOR(95

CI)b

119875b

rs2228001(HWE=0948)

AA

99(391

3)218(410

5)10

010

0AC

118(4664

)245(4614)

106(077ndash14

7)0722

105(076

ndash145)

0769

CC36

(1423)

68(1281)

117(073ndash18

6)0521

115(072ndash18

4)0555

Additiv

e0807

108(086ndash

134)

0520

107(086ndash

133)

0563

Dom

inant

154(6087)

313(5895)

0608

108(080ndash

147)

0608

107(079ndash

146)

0656

Recessive

217(8577)

463(8719)

0583

113(073ndash17

5)0583

112(073ndash174)

060

4rs222800

0(H

WE=0988)

CC111(4387)

205(3861)

100

100

CT108(4269)

250(470

8)080

(058ndash110)

0170

080

(058ndash110)

0166

TT34

(1344

)76

(1431

)083

(052

ndash132

)0422

084

(052

ndash133

)0453

Additiv

e0368

088

(071ndash10

9)0244

088

(071ndash110)

0257

Dom

inant

142(5613)

326(613

9)0160

080

(059ndash

109)

0160

081

(059ndash

109)

0162

Recessive

219(8656)

455(8569)

0742

093

(060ndash

144)

0742

094

(061ndash14

6)0785

rs2229090(H

WE=0994)

GG

99(391

3)191(3597)

100

100

GC

105(415

0)255(4802)

079

(057ndash111)

0175

079

(057ndash110)

0169

CC49

(1937

)85

(1601)

111(073ndash171)

0626

111(073ndash171)

0622

Additiv

e0204

100(081ndash12

4)0971

100(081ndash12

4)0972

Dom

inant

154(6087)

340(6403)

0392

087

(064ndash

119)

0392

087

(064ndash

119)

0384

Recessive

204(8063)

446(8399)

0243

126(085ndash18

6)0243

126(086ndash

187)

0237

a 1205942testforg

enotyped

istrib

utions

betweenneurob

lasto

mac

ases

andcancer-fr

eecontrols

b Adjustedfora

geandgend

er


Table2Stratifi

catio

nanalysisofris

kgeno

typesa

ndneurob

lasto

mas

usceptibility

Varia

bles

rs2228001(casescontrols)

Adjuste

dOR

Prs2228000(casescon

trols)

Adjuste

dORa

119875a

rs2229090(casescon

trols)

Adjuste

dORa

119875a

AA

ACC

C(95

CI)

CCCT

TT

(95

CI)

GG

GCCC

(95

CI)

Agem

onth

le18

4710

25313

1087

(054ndash

139)

0557

1737

8319

6093

(049ndash

174)

0811

4379

5715

4068

(042ndash110)

0119

gt18

5211

61011

8212

4(082ndash18

6)0304

1739

136259

120(065ndash220)

0559

5611

29718

610

4(069ndash

155)

0862

Sex Fe

males

4695

5513

8081

(050ndash

130)

0383

1731

84202

075

(039

ndash143)

0382

3789

6414

410

8(066ndash

174)

0771

Males

5312

39917

513

1(087ndash197)

0190

1745

135253

140(077ndash255)

0270

6210

29019

6075

(050ndash

113)

0167

Siteso

forig

inAd

renalgland

18218

28313

104(056ndash

194)

0896

476

42455

168(058ndash

485)

0335

1819

128340

088

(047ndash16

4)0689

Retro

periton

eal

37218

48313

090

(057ndash14

3)0654

1476

71455

084

(045ndash15

7)0582

3619

149340

075

(047ndash12

0)0226

Mediastinum

30218

59313

138(086ndash

222)

0181

1276

77455

108(056ndash

208)

0825

3519

154340

087

(055ndash13

8)0546

Other

sites

11218

14313

092

(041ndash207)

0837

376

33455

131(038ndash4

50)

0670

8191

17340

116(049ndash

275)

0734

Clinicalstages

I+II+4s

50218

73313

103(069ndash

154)

0872

1576

108455

124(068ndash224)

0486

5119

172340

079

(053

ndash118

)0257

III+

IV46

218

75313

109(072ndash16

4)0680

1776

104455

095

(054ndash

169)

0863

4719

174340

090

(060ndash

135)

0599

a Adjustedfora

geandgend

er


4 Discussion

In this hospital-based study comprising 256 cases and 531controls none of the three XPC gene polymorphisms wasassociated with neuroblastoma risk when compared to thereference genotypes To the best of our knowledge this is thefirst investigation looking into the relationship between XPCgene polymorphisms and neuroblastoma susceptibility in aChinese Han population

XPC gene (httpwwwncbinlmnihgovgene7508) islocated on chromosome 3p251 with 18 exons which encodesa component of the NER XPC plays a distinctively vital rolein the early stages of global genome NER XPC and UVexcision repair protein RAD23 homolog B (HR23B) form theXPC-HR23B complex which recognizes DNA damage andinitiates NER in mammalian cells thereby protecting againstcancer [20ndash25 41 42]

XPC gene polymorphisms are involved in the differenttypes of cancer Among them rs2228001 AgtC and rs2228000CgtT were widely investigated Paszkowska-Szczur et al [26]genotyped 714 melanoma cases and 1841 healthy controlsto evaluate the relationship between 94 SNPs within theseven XP genes (XPAndashXPG) and the melanoma risk in aPolish population They found that XPC rs2228000 CT andTT genotypes were significantly associated with decreasedmelanoma risk when compared with the reference genotypeMoreover they observed that rs2228000 CT genotype andrs2228000 TT genotype were associated with decreasedcolorectal cancer risk compared to the CC genotype in 758colorectal cancer patients and the same number of controls[27] In a study conducted in Malaysia with 255 colorectalcancer patients and 255 controls Ahmad Aizat et al [28]found that XPC gene rs2228000 GG genotype was associatedwith an increased colorectal cancer risk However there isno previous study investigating the association between XPCgene polymorphisms and neuroblastoma susceptibility [43]Our study fits the niche and it found no association betweenthe three SNPs ofXPC gene and neuroblastomaHowever thenegative results might result from the relatively small samplesize in this study although it was the largest study regardingthe Chinese children to date

5 Conclusions

In summary all the three XPC gene polymorphisms(rs2228001 AgtC rs2228000 CgtT and rs2229090 GgtC) maynot associate with neuroblastoma risk in the Chinese Hanpopulation However further studies with larger sample sizeand different ethnicities should be carried out to verify ourresults

Abbreviations

GWAS Genome-wide association studySNP Single nucleotide polymorphismXPC Xeroderma pigmentosum complementation CNER Nucleotide excision repair pathwayXP Xeroderma pigmentosumOR Odds ratioCI Confidence interval

Competing Interests

The authors declare that there are no competing interestsregarding the publication of this paper

Authorsrsquo Contributions

All authors contributed significantly to this work JintaoZheng Ruizhong Zhang Fenghua Wang and Tianyou Yangperformed the research study and collected the data Jing Heand Jintao Zheng analyzed the data Jing He and Huimin Xiadesigned the research study Jintao Zheng Jinhong Zhu andJing He wrote the paper and Jing He prepared all the tablesAll authors reviewed the manuscript In addition all authorsapproved the final draft

Acknowledgments

This work was supported by the grant of State Clinical KeySpecialty Construction Project (Pediatric Surgery) 2013 (noGJLCZD1301) the grant of Clinical Medicine Research andTransformationCenter of Brain Injury in Premature Infant inGuangzhou (no 520101-2150092) and the National NaturalScience Foundation of China (no 81502046) The authorsthank Yanlu Tong and Hezhen Wang for their assistance inDNA extraction and medical histories information collec-tion

References

[1] M A Smith N L Seibel S F Altekruse et al ldquoOutcomes forchildren and adolescents with cancer challenges for the twenty-first centuryrdquo Journal of Clinical Oncology vol 28 no 15 pp2625ndash2634 2010

[2] J M Maris M D Hogarty R Bagatell and S L CohnldquoNeuroblastomardquoThe Lancet vol 369 no 9579 pp 2106ndash21202007

[3] G M Brodeur J Pritchard F Berthold et al ldquoRevisions of theinternational criteria for neuroblastoma diagnosis staging andresponse to treatmentrdquo Journal of Clinical Oncology vol 11 no8 pp 1466ndash1477 1993

[4] P-P Bao K Li C-X Wu et al ldquoRecent incidences and trendsof childhood malignant solid tumors in Shanghai 2002ndash2010rdquoZhonghua Er Ke Za Zhi vol 51 no 4 pp 288ndash294 2013

[5] F Berthold J Boos S Burdach et al ldquoMyeloablative megather-apy with autologous stem-cell rescue versus oral maintenancechemotherapy as consolidation treatment in patients with high-risk neuroblastoma a randomised controlled trialrdquo The LancetOncology vol 6 no 9 pp 649ndash658 2005

[6] J M Maris ldquoRecent advances in neuroblastomardquo The NewEngland Journal of Medicine vol 362 no 23 pp 2154ndash22112010

[7] K A Frazer S S Murray N J Schork and E J Topol ldquoHumangenetic variation and its contribution to complex traitsrdquoNatureReviews Genetics vol 10 no 4 pp 241ndash251 2009

[8] T A Manolio ldquoGenomewide association studies and assess-ment of the risk of diseaserdquo The New England Journal ofMedicine vol 363 no 2 pp 166ndash176 2010

[9] J MMaris Y PMosse J P Bradfield et al ldquoChromosome 6p22locus associated with clinically aggressive neuroblastomardquo The


New England Journal of Medicine vol 358 no 24 pp 2585ndash2593 2008

[10] S J Diskin M Capasso R W Schnepp et al ldquoCommon varia-tion at 6q16withinHACE1 and LIN28B influences susceptibilityto neuroblastomardquo Nature Genetics vol 44 no 10 pp 1126ndash1130 2012

[11] M Capasso M Devoto C Hou et al ldquoCommon variationsin BARD1 influence susceptibility to high-risk neuroblastomardquoNature Genetics vol 41 no 6 pp 718ndash723 2009

[12] K Wang S J Diskin H Zhang et al ldquoIntegrative genomicsidentifies LMO1 as a neuroblastoma oncogenerdquoNature vol 469no 7329 pp 216ndash220 2011

[13] L B Nguyễn S J Diskin M Capasso et al ldquoPhenotyperestricted genome-wide association study using a gene-centricapproach identifies three low-risk neuroblastoma susceptibilitylocirdquo PLoS Genetics vol 7 no 3 Article ID e1002026 2011

[14] WHan Y Zhou R Zhong et al ldquoFunctional polymorphisms inFASFASL system increase the risk of neuroblastoma inChinesepopulationrdquo PLoS ONE vol 8 no 8 Article ID e71656 2013

[15] M Capasso S Diskin F Cimmino et al ldquoCommon geneticvariants inNEFL influence gene expression andNeuroblastomariskrdquo Cancer Research vol 74 no 23 pp 6913ndash6924 2014

[16] Y Jin HWangWHan et al ldquoSingle nucleotide polymorphismrs11669203 in TGFBR3L is associated with the risk of neuroblas-toma in a Chinese populationrdquo Tumor Biology vol 37 no 3 pp3739ndash3747 2016

[17] R D Wood M Mitchell J Sgouros and T Lindahl ldquoHumanDNA repair genesrdquo Science vol 291 no 5507 pp 1284ndash12892001

[18] E C Friedberg ldquoHow nucleotide excision repair protectsagainst cancerrdquo Nature Reviews Cancer vol 1 no 1 pp 22ndash332001

[19] J E Cleaver ldquoDefective repair replication of DNA in xerodermapigmentosumrdquo Nature vol 218 no 5142 pp 652ndash656 1968

[20] T Riedl F Hanaoka and J-M Egly ldquoThe comings and goingsof nucleotide excision repair factors on damaged DNArdquo EMBOJournal vol 22 no 19 pp 5293ndash5303 2003

[21] M Volker M J Mone P Karmakar et al ldquoSequential assemblyof the nucleotide excision repair factors in vivordquoMolecular Cellvol 8 no 1 pp 213ndash224 2001

[22] J H J Hoeijmakers ldquoGenome maintenance mechanisms forpreventing cancerrdquoNature vol 411 no 6835 pp 366ndash374 2001

[23] J De Boer and J H J Hoeijmakers ldquoNucleotide excision repairand human syndromesrdquo Carcinogenesis vol 21 no 3 pp 453ndash460 2000

[24] W L de Laat N G J Jaspers and J H J HoeijmakersldquoMolecular mechanism of nucleotide excision repairrdquo Genes ampDevelopment vol 13 no 7 pp 768ndash785 1999

[25] R D Wood ldquoNucleotide excision repair in mammalian cellsrdquoJournal of Biological Chemistry vol 272 no 38 pp 23465ndash23468 1997

[26] K Paszkowska-Szczur R J Scott P Serrano-Fernandez et alldquoXeroderma pigmentosum genes and melanoma riskrdquo Interna-tional Journal of Cancer vol 133 no 5 pp 1094ndash1100 2013

[27] K Paszkowska-Szczur R J Scott B Gorski et al ldquoPolymor-phisms in nucleotide excision repair genes and susceptibility tocolorectal cancer in the Polish populationrdquo Molecular BiologyReports vol 42 no 3 pp 755ndash764 2015

[28] A A Ahmad Aizat M S Siti Nurfatimah M M Aminudinand R Ankathil ldquoXPC Lys939Gln polymorphism smoking and

risk of sporadic colorectal cancer among Malaysiansrdquo WorldJournal of Gastroenterology vol 19 no 23 pp 3623ndash3628 2013

[29] Y Yoshino S Takeuchi T Katoh and Y Kuroda ldquoXPC intron11CApolymorphism as a risk factor for prostate cancerrdquoEnviron-mental Health and Preventive Medicine vol 21 no 2 pp 100ndash104 2016

[30] B Wang Q Xu H W Yang L P Sun and Y Yuan ldquoTheassociation of six polymorphisms of five genes involved in threesteps of nucleotide excision repair pathways with hepatocellularcancer riskrdquo Oncotarget vol 7 no 15 pp 20357ndash20367 2016

[31] C Mei M Hou S Guo et al ldquoPolymorphisms in DNA repairgenes of XRCC1 XPA XPC XPD and associations with lungcancer risk in Chinese peoplerdquo Thoracic Cancer vol 5 no 3pp 232ndash242 2014

[32] J Liu L Sun Q Xu et al ldquoAssociation of nucleotide excisionrepair pathway gene polymorphisms with gastric cancer andatrophic gastritis risksrdquoOncotarget vol 7 no 6 pp 6972ndash69832016

[33] JHe FWang J Zhu et al ldquoAssociation of potentially functionalvariants in the XPG gene with neuroblastoma risk in a Chinesepopulationrdquo Journal of Cellular andMolecularMedicine vol 20pp 1481ndash1490 2016

[34] J He T Yang R Zhang et al ldquoPotentially functional poly-morphisms in the LIN28B gene contribute to neuroblastomasusceptibility in Chinese childrenrdquo Journal of Cellular andMolecular Medicine vol 20 no 8 pp 1534ndash1541 2016

[35] J HeW Zhong J Zeng et al ldquoLMO1 gene polymorphisms con-tribute to decreased neuroblastoma susceptibility in a SouthernChinese populationrdquoOncotarget vol 7 no 16 pp 22770ndash227782016

[36] R Zhang Y Zou J Zhu et al ldquoThe association betweenGWAS-identified BARD1 gene SNPS and neuroblastoma susceptibilityin a Southern Chinese populationrdquo International Journal ofMedical Sciences vol 13 no 2 pp 133ndash138 2016

[37] J He R Zhang Y Zou et al ldquoEvaluation of GWAS-identifiedSNPs at 6p22 with neuroblastoma susceptibility in a Chinesepopulationrdquo Tumor Biology vol 37 no 2 pp 1635ndash1639 2016

[38] R X Hua Z J Zhuo G P Shen et al ldquoPolymorphisms in theXPC gene and gastric cancer susceptibility in a Southern Chi-nese populationrdquo Journal of OncoTargets andTherapy vol 9 pp5513ndash5519 2016

[39] J He L-X Qiu M-Y Wang et al ldquoPolymorphisms in the XPGgene and risk of gastric cancer in Chinese populationsrdquoHumanGenetics vol 131 no 7 pp 1235ndash1244 2012

[40] J He M-Y Wang L-X Qiu et al ldquoGenetic variations ofmTORC1 genes and risk of gastric cancer in an Eastern ChinesepopulationrdquoMolecular Carcinogenesis vol 52 supplement 1 ppE70ndashE79 2013

[41] M Akita Y-S Tak T Shimura et al ldquoSUMOylation of xero-derma pigmentosum group C protein regulates DNA dam-age recognition during nucleotide excision repairrdquo ScientificReports vol 5 Article ID 10984 2015

[42] K Sugasawa J M Y Ng C Masutani et al ldquoXerodermapigmentosum group C protein complex is the initiator of globalgenome nucleotide excision repairrdquoMolecular Cell vol 2 no 2pp 223ndash232 1998

[43] J He T-Y Shi M-L Zhu M-YWang Q-X Li and Q-YWeildquoAssociations of Lys939Gln and Ala499Val polymorphisms ofthe XPC gene with cancer susceptibility a meta-analysisrdquo Inter-national Journal of Cancer vol 133 no 8 pp 1765ndash1775 2013

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014


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The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

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ObesityJournal of



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Research and TreatmentAIDS


Gastroenterology Research and Practice


Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom


Several gene polymorphisms are significantly associated withthe risk of neuroblastoma such as LIN28B [10] HACE1 [10]BARD1 [11] LMO1 [12] HSD17B12 [13] DDX4 [13] IL31RA[13] and DUSP12 [13] which have been discovered by meansof GWAS Besides several genes such as FAS [14] FASL[14] NEFL [15] and TGFBR3L [16] have been discovered bycandidate gene approach

Nucleotide excision repair (NER) can preserve the hered-itary integrity and stability of genomic DNA through elim-inating DNA damage generated by exogenous agents (iemutagenic and carcinogenic substances) and photoproductscaused by sunlight exposure [17 18] In humans hereditarydefects of genomic DNA in NER are involved in several auto-somal recessive diseases Xeroderma pigmentosum (XP) ischaracterized by a strong predisposition to skin carcinomasHowever it was not until 1968 that Cleaver [19] identified thatXP could result from the defects in NER NER is a complexbiochemical process that involves hundreds of componentsin human cells including the seven XP-related genes (XPAXPB XPC XPD XPE XPF and XPG) XPC protein is animportant DNA damage recognition protein and an initiatorof the NER process that defends against cancer [20ndash25]

Some XPC gene SNPs have been identified to be impli-cated in melanoma [26] colorectal cancer [27 28] prostatecancer [29] hepatocellular cancer [30] lung cancer [31]and gastric cancer [32] However the association betweenXPC gene polymorphisms and neuroblastoma risk has notbeen studied In view of this we investigated the relation-ship between XPC gene polymorphisms (rs2228001 AgtCrs2228000 CgtT and rs2229090 GgtC) and neuroblastomasusceptibility in a Chinese Han population making use of atotal of 787 participants (256 cases and 531 controls)

2 Materials and Methods

21 Study Subjects A total of 256 newly diagnosed andhistopathologically confirmed neuroblastoma cases wererecruited from the Guangzhou Women and Childrenrsquos Med-ical Center between February 2010 and November 2015 [33ndash35] During the same period 531 healthy children were ran-domly selected as the age- and gender-matched controls afterreceiving routine physical examination [35ndash37] Both thecases and the controls were unassociated ethnic Chinese Hanindividuals The study was granted permission by the Insti-tutional Review Board of GuangzhouWomen and ChildrenrsquosMedical Center Demographic factors and medical historieswere gathered through the structured questionnairesWritteninformed consent was received from the guardians of eachchild participant Each participant donated 2mL of blood forgenomic DNA extraction

22 Polymorphism Analysis The selection criteria weredescribed previously in terms of the minor allele frequencyputative functional potentials and low linkage disequilib-rium [38] Based on the criteria three potentially functionalSNPs (rs2228001 AgtC rs2228000 CgtT and rs2229090 GgtC)were selected s2228001 AgtC (Lys939Gln) and rs2228000CgtT (Val499Arg) are nonsynonymous SNPs and they havebeen widely investigated in various types of cancer XPC

rs2229090 GgtC is 31015840UTR polymorphism within miRNAbinding site Genomic DNA was extracted from 2mL ofperipheral blood sample using the TIANamp Blood DNAKit (TianGen Biotech Co Ltd Beijing China) following themanufacturerrsquos instructions Qualified DNA samples werediluted to 10 ng120583L and loaded in 96-well plates Then thesethree SNPs were genotyped using Taqman real-time PCRmethod as described previously [39 40]

23 Statistical Analysis 1205942 test was used to evaluate the dif-ferences in the frequency distributions of the demographicsand genotypes between the neuroblastoma cases and thecontrols Hardy-Weinberg equilibrium (HWE) was tested inthe controls by the goodness-of-fit chi-squared test Oddsratios (ORs) and 95 confidence intervals (CIs) were usedto assess the correlations between the three polymorphismsand neuroblastoma susceptibility with the unconditionalmultivariate logistic regression analysis 119875 value lt 005 wasconsidered as statistically significant All statistical tests weretwo-sided and analyzed using SAS software (version 91 SASInstitute Cary NC)

3 Results

31 Frequency Distribution of Selected Characteristics Asdisplayed in Supplemental Table 1 in SupplementaryMaterialavailable online at httpdxdoiorg10115520162932049the average age of the cases was 3087 plusmn 2645 monthsand 2973 plusmn 2486 months for the controls No significantdifferences were observed in terms of age (119875 = 0239) andgender (119875 = 0333) between the case and the control groupsAccording to the INSS standard [3] 54 65 44 77 and 9patients developed clinical stages I II III and IV and 4sneuroblastoma respectively Among these cases 46 lesionsoccurred in adrenal gland 87 in retroperitoneal region and90 in mediastinum

32 XPC Gene Polymorphisms and Neuroblastoma Suscepti-bility In the current study 253 cases and 531 controls weresuccessfully genotypedThe genotype frequencies of the threepolymorphisms are shown in Table 1 The observed genotypefrequencies of the three SNPs were in accordance with HWEin the control subjects (119875 = 0948 for rs2228001 AgtCpolymorphism 119875 = 0988 for rs2228000 CgtT polymor-phism and 119875 = 0994 for rs2229090 GgtC polymorphism)There is no significant association between rs2228001 AgtCpolymorphism and neuroblastoma susceptibility Similarresults were found for rs2228000 CgtT and rs2229090 GgtCpolymorphisms

33 Stratification Analysis of XPC Gene Polymorphisms withNeuroblastoma Susceptibility Stratified analyses were con-ducted regarding age gender sites of origin and clinicalstages to assess the association of the three selected polymor-phisms with the risk of neuroblastoma (Table 2) Howeverno significant association was identified for any of the threepolymorphisms


Table1Genotypea

ndallelefre

quencies

ofthethree

selected

polymorph

ismsa

ndneurob

lasto

mas

usceptibilityin

aChinese

popu

lation

Genotype

Cases(119873=253)

Con

trols(119873=531)

119875a

Crud

eOR(95

CI)

119875Ad

juste

dOR(95

CI)b

119875b

rs2228001(HWE=0948)

AA

99(391

3)218(410

5)10

010

0AC

118(4664

)245(4614)

106(077ndash14

7)0722

105(076

ndash145)

0769

CC36

(1423)

68(1281)

117(073ndash18

6)0521

115(072ndash18

4)0555

Additiv

e0807

108(086ndash

134)

0520

107(086ndash

133)

0563

Dom

inant

154(6087)

313(5895)

0608

108(080ndash

147)

0608

107(079ndash

146)

0656

Recessive

217(8577)

463(8719)

0583

113(073ndash17

5)0583

112(073ndash174)

060

4rs222800

0(H

WE=0988)

CC111(4387)

205(3861)

100

100

CT108(4269)

250(470

8)080

(058ndash110)

0170

080

(058ndash110)

0166

TT34

(1344

)76

(1431

)083

(052

ndash132

)0422

084

(052

ndash133

)0453

Additiv

e0368

088

(071ndash10

9)0244

088

(071ndash110)

0257

Dom

inant

142(5613)

326(613

9)0160

080

(059ndash

109)

0160

081

(059ndash

109)

0162

Recessive

219(8656)

455(8569)

0742

093

(060ndash

144)

0742

094

(061ndash14

6)0785

rs2229090(H

WE=0994)

GG

99(391

3)191(3597)

100

100

GC

105(415

0)255(4802)

079

(057ndash111)

0175

079

(057ndash110)

0169

CC49

(1937

)85

(1601)

111(073ndash171)

0626

111(073ndash171)

0622

Additiv

e0204

100(081ndash12

4)0971

100(081ndash12

4)0972

Dom

inant

154(6087)

340(6403)

0392

087

(064ndash

119)

0392

087

(064ndash

119)

0384

Recessive

204(8063)

446(8399)

0243

126(085ndash18

6)0243

126(086ndash

187)

0237

a 1205942testforg

enotyped

istrib

utions

betweenneurob

lasto

mac

ases

andcancer-fr

eecontrols

b Adjustedfora

geandgend

er


Table2Stratifi

catio

nanalysisofris

kgeno

typesa

ndneurob

lasto

mas

usceptibility

Varia

bles


Adjuste

dOR

Prs2228000(casescon

trols)

Adjuste

dORa

119875a

rs2229090(casescon

trols)

Adjuste

dORa

119875a

AA

ACC

C(95

CI)

CCCT

TT

(95

CI)

GG

GCCC

(95

CI)

Agem

onth

le18

4710

25313

1087

(054ndash

139)

0557

1737

8319

6093

(049ndash

174)

0811

4379

5715

4068

(042ndash110)

0119

gt18

5211

61011

8212

4(082ndash18

6)0304

1739

136259

120(065ndash220)

0559

5611

29718

610

4(069ndash

155)

0862

Sex Fe

males

4695

5513

8081

(050ndash

130)

0383

1731

84202

075

(039

ndash143)

0382

3789

6414

410

8(066ndash

174)

0771

Males

5312

39917

513

1(087ndash197)

0190

1745

135253

140(077ndash255)

0270

6210

29019

6075

(050ndash

113)

0167

Siteso

forig

inAd

renalgland

18218

28313

104(056ndash

194)

0896

476

42455

168(058ndash

485)

0335

1819

128340

088

(047ndash16

4)0689

Retro

periton

eal

37218

48313

090

(057ndash14

3)0654

1476

71455

084

(045ndash15

7)0582

3619

149340

075

(047ndash12

0)0226

Mediastinum

30218

59313

138(086ndash

222)

0181

1276

77455

108(056ndash

208)

0825

3519

154340

087

(055ndash13

8)0546

Other

sites

11218

14313

092

(041ndash207)

0837

376

33455

131(038ndash4

50)

0670

8191

17340

116(049ndash

275)

0734

Clinicalstages

I+II+4s

50218

73313

103(069ndash

154)

0872

1576

108455

124(068ndash224)

0486

5119

172340

079

(053

ndash118

)0257

III+

IV46

218

75313

109(072ndash16

4)0680

1776

104455

095

(054ndash

169)

0863

4719

174340

090

(060ndash

135)

0599

a Adjustedfora

geandgend

er


4 Discussion




5 Conclusions


Abbreviations


Competing Interests




Acknowledgments


References




















































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Table1Genotypea

ndallelefre

quencies

ofthethree

selected

polymorph

ismsa

ndneurob

lasto

mas

usceptibilityin

aChinese

popu

lation

Genotype

Cases(119873=253)

Con

trols(119873=531)

119875a

Crud

eOR(95

CI)

119875Ad

juste

dOR(95

CI)b

119875b

rs2228001(HWE=0948)

AA

99(391

3)218(410

5)10

010

0AC

118(4664

)245(4614)

106(077ndash14

7)0722

105(076

ndash145)

0769

CC36

(1423)

68(1281)

117(073ndash18

6)0521

115(072ndash18

4)0555

Additiv

e0807

108(086ndash

134)

0520

107(086ndash

133)

0563

Dom

inant

154(6087)

313(5895)

0608

108(080ndash

147)

0608

107(079ndash

146)

0656

Recessive

217(8577)

463(8719)

0583

113(073ndash17

5)0583

112(073ndash174)

060

4rs222800

0(H

WE=0988)

CC111(4387)

205(3861)

100

100

CT108(4269)

250(470

8)080

(058ndash110)

0170

080

(058ndash110)

0166

TT34

(1344

)76

(1431

)083

(052

ndash132

)0422

084

(052

ndash133

)0453

Additiv

e0368

088

(071ndash10

9)0244

088

(071ndash110)

0257

Dom

inant

142(5613)

326(613

9)0160

080

(059ndash

109)

0160

081

(059ndash

109)

0162

Recessive

219(8656)

455(8569)

0742

093

(060ndash

144)

0742

094

(061ndash14

6)0785

rs2229090(H

WE=0994)

GG

99(391

3)191(3597)

100

100

GC

105(415

0)255(4802)

079

(057ndash111)

0175

079

(057ndash110)

0169

CC49

(1937

)85

(1601)

111(073ndash171)

0626

111(073ndash171)

0622

Additiv

e0204

100(081ndash12

4)0971

100(081ndash12

4)0972

Dom

inant

154(6087)

340(6403)

0392

087

(064ndash

119)

0392

087

(064ndash

119)

0384

Recessive

204(8063)

446(8399)

0243

126(085ndash18

6)0243

126(086ndash

187)

0237

a 1205942testforg

enotyped

istrib

utions

betweenneurob

lasto

mac

ases

andcancer-fr

eecontrols

b Adjustedfora

geandgend

er


Table2Stratifi

catio

nanalysisofris

kgeno

typesa

ndneurob

lasto

mas

usceptibility

Varia

bles


Adjuste

dOR

Prs2228000(casescon

trols)

Adjuste

dORa

119875a

rs2229090(casescon

trols)

Adjuste

dORa

119875a

AA

ACC

C(95

CI)

CCCT

TT

(95

CI)

GG

GCCC

(95

CI)

Agem

onth

le18

4710

25313

1087

(054ndash

139)

0557

1737

8319

6093

(049ndash

174)

0811

4379

5715

4068

(042ndash110)

0119

gt18

5211

61011

8212

4(082ndash18

6)0304

1739

136259

120(065ndash220)

0559

5611

29718

610

4(069ndash

155)

0862

Sex Fe

males

4695

5513

8081

(050ndash

130)

0383

1731

84202

075

(039

ndash143)

0382

3789

6414

410

8(066ndash

174)

0771

Males

5312

39917

513

1(087ndash197)

0190

1745

135253

140(077ndash255)

0270

6210

29019

6075

(050ndash

113)

0167

Siteso

forig

inAd

renalgland

18218

28313

104(056ndash

194)

0896

476

42455

168(058ndash

485)

0335

1819

128340

088

(047ndash16

4)0689

Retro

periton

eal

37218

48313

090

(057ndash14

3)0654

1476

71455

084

(045ndash15

7)0582

3619

149340

075

(047ndash12

0)0226

Mediastinum

30218

59313

138(086ndash

222)

0181

1276

77455

108(056ndash

208)

0825

3519

154340

087

(055ndash13

8)0546

Other

sites

11218

14313

092

(041ndash207)

0837

376

33455

131(038ndash4

50)

0670

8191

17340

116(049ndash

275)

0734

Clinicalstages

I+II+4s

50218

73313

103(069ndash

154)

0872

1576

108455

124(068ndash224)

0486

5119

172340

079

(053

ndash118

)0257

III+

IV46

218

75313

109(072ndash16

4)0680

1776

104455

095

(054ndash

169)

0863

4719

174340

090

(060ndash

135)

0599

a Adjustedfora

geandgend

er


4 Discussion




5 Conclusions


Abbreviations


Competing Interests




Acknowledgments


References




















































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Table2Stratifi

catio

nanalysisofris

kgeno

typesa

ndneurob

lasto

mas

usceptibility

Varia

bles


Adjuste

dOR

Prs2228000(casescon

trols)

Adjuste

dORa

119875a

rs2229090(casescon

trols)

Adjuste

dORa

119875a

AA

ACC

C(95

CI)

CCCT

TT

(95

CI)

GG

GCCC

(95

CI)

Agem

onth

le18

4710

25313

1087

(054ndash

139)

0557

1737

8319

6093

(049ndash

174)

0811

4379

5715

4068

(042ndash110)

0119

gt18

5211

61011

8212

4(082ndash18

6)0304

1739

136259

120(065ndash220)

0559

5611

29718

610

4(069ndash

155)

0862

Sex Fe

males

4695

5513

8081

(050ndash

130)

0383

1731

84202

075

(039

ndash143)

0382

3789

6414

410

8(066ndash

174)

0771

Males

5312

39917

513

1(087ndash197)

0190

1745

135253

140(077ndash255)

0270

6210

29019

6075

(050ndash

113)

0167

Siteso

forig

inAd

renalgland

18218

28313

104(056ndash

194)

0896

476

42455

168(058ndash

485)

0335

1819

128340

088

(047ndash16

4)0689

Retro

periton

eal

37218

48313

090

(057ndash14

3)0654

1476

71455

084

(045ndash15

7)0582

3619

149340

075

(047ndash12

0)0226

Mediastinum

30218

59313

138(086ndash

222)

0181

1276

77455

108(056ndash

208)

0825

3519

154340

087

(055ndash13

8)0546

Other

sites

11218

14313

092

(041ndash207)

0837

376

33455

131(038ndash4

50)

0670

8191

17340

116(049ndash

275)

0734

Clinicalstages

I+II+4s

50218

73313

103(069ndash

154)

0872

1576

108455

124(068ndash224)

0486

5119

172340

079

(053

ndash118

)0257

III+

IV46

218

75313

109(072ndash16

4)0680

1776

104455

095

(054ndash

169)

0863

4719

174340

090

(060ndash

135)

0599

a Adjustedfora

geandgend

er


4 Discussion




5 Conclusions


Abbreviations


Competing Interests




Acknowledgments


References




















































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















4 Discussion




5 Conclusions


Abbreviations


Competing Interests




Acknowledgments


References




















































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of


























































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















research article lack of associations between xpc gene...

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