Research Article Involvement of Fibroblast Growth Factor...

Hindawi Publishing CorporationDisease MarkersVolume 35 (2013) Issue 6 Pages 869ndash875httpdxdoiorg1011552013792941

Research ArticleInvolvement of Fibroblast Growth Factor Receptor Genes inBenign Prostate Hyperplasia in a Korean Population

Hae Jeong Park1 Su Kang Kim1 Jong Woo Kim1

Sang Hyub Lee2 Koo Han Yoo2 and Joo-Ho Chung1

1 Kohwang Medical Research Institute School of Medicine Kyung Hee University 26 Kyunghee-daeroDongdaemun-gu Seoul 130-701 Republic of Korea

2Department of Urology School of Medicine Kyung Hee University 26 Kyunghee-daero Dongdaemun-guSeoul 130-701 Republic of Korea

Correspondence should be addressed to Joo-Ho Chung hjpark17gmailcom

Received 3 September 2013 Revised 1 November 2013 Accepted 15 November 2013

Academic Editor Lance A Liotta

Copyright copy 2013 Hae Jeong Park et alThis 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

Fibroblast growth factors (FGFs) and their receptors (FGFRs) have been implicated in prostate growth and are overexpressedin benign prostatic hyperplasia (BPH) In this study we investigated whether single nucleotide polymorphisms (SNPs) of theFGFR genes (FGFR1 and FGFR2) were associated with BPH and its clinical phenotypes in a population of Korean men Wegenotyped four SNPs in the exons of FGFR1 and FGFR2 (rs13317 in FGFR1 rs755793 rs1047100 and rs3135831 in FGFR2) usingdirect sequencing in 218 BPH patients and 213 control subjects No SNPs of FGFR1 or FGFR2 genes were associated with BPHHowever analysis according to clinical phenotypes showed that rs1047100 of FGFR2 was associated with prostate volume in BPHin the dominant model (GAAA versus GG P = 0010) In addition a significant association was observed between rs13317 ofFGFR1 and international prostate symptom score (IPSS) in the additive (TC versus CC versus TT P = 00022) and dominantmodels (TCCC versus TT P = 0005) Allele frequency analysis also showed significant association between rs13317 and IPSS (P =0005) These results suggested that FGFR genes could be related to progression of BPH

1 Introduction

Benign prostatic hyperplasia (BPH) is the most commonurological problem associated with aging in men One-quarter of men in their 50s one-third in their 60s and halfof men older than 80 have BPH [1] BPH is characterizedby hyperplasia of prostatic stromal and epithelial cells andit manifests as a severe obstruction in urinary flow with dis-comfort and painThe pathogenesis of BPH is not completelyunderstood however the most significant risk factors forthe development of BPH are androgen level and aging [2]Growth factors and their receptors including members ofthe fibroblast growth factor (FGF) family insulin-like growthfactor (IGF) family epithelial growth factor (EGF) familyand transforming growth factor 120573 (TGFB) which regulatethe growth of prostatic stromal and epithelial cells are alsoinvolved in the pathogenesis of BPH [3ndash5]

Genetic strategies have been used over the past fewdecades to investigate BPH In particular studies have shown

that polymorphisms of growth factors and their receptorgenes are associated with BPH Indeed previous studieshave reported that a codon 10 polymorphism in TGFB1 wasassociated with the development of BPH in Japanese [6]and Iranian populations [7] suggesting the importance ofthe TGF pathway in the development of prostatic diseasesMullan et al [8] reported a significant association of acodon 10 polymorphism in TGFB1 with treatment for BPHand an association of a CA-repeat polymorphismin EGFRwith international prostate symptom score (IPSS) in BPHMoreover a CA-repeat polymorphism of 19-allele in IGF1appears to increase the risk of BPH with a gene dosageeffect in the Japanese population [9] However despite awidespread consensus on the involvement of growth factorsin prostatic growth attempts to address growth factor genepolymorphisms in patients with BPH have been limited

FGFs are involved inmultiple biological processes such asdifferentiation motility and proliferation and mediate their

870 Disease Markers

cellular responses by activating a family of four receptortyrosine kinases FGFR1 through FGFR4 [10] FGFRs arepresent in various organs In the humanprostate FGFR1-3 areabundantly expressed in the prostate stroma andor epithe-lium [11ndash14] and FGFR4 has relatively low expression in theepithelium [15] In the prostate FGFRs play an importantrole in prostate organogenesis and perturbations in FGFRexpression have been potently implicated in prostatic disease[11] Previous studies have reported that overexpression ofFGFRs is important in the development of BPH [16ndash18]Indeed FGFR1 was observed to be increased in the stromaof BPH patients compared to that from normal prostates[16 17] and increased expression of FGFR2 has been detectedin BPH [18] Furthermore although no studies have reportedchanges in expression of FGFR4 in BPH a genetic studyshowed a significant association between FGFR4 polymor-phisms and BPH in a Japanese population [19] In light ofthese findings we postulated that FGFRs may be involvedin the pathogenesis of BPH In this study we investigatedthe genetic associations between BPH and the FGFR genes(FGFR1 and FGFR2) in Korean BPH patients by analyz-ing single nucleotide polymorphisms (SNPs) of the FGFRgenes

2 Methods

21 Subjects A total of 233 male patients with BPH (meanage plusmn standard deviation 6577 plusmn 946 years) and 213 malecontrol subjects (mean age 6189 plusmn 826 years) were enrolledAll patients with BPH were from Kyung Hee UniversityHospital between January 2002 and December 2008 and inall patients lower urinary tract symptoms were quantifiedusing IPSS Uroflowmetry was performed to measure peakurinary flow rate (119876max) for all patients Serum prostate-specific antigen (PSA) level wasmeasured in all BPHpatientsPatients with serumPSA level more than 4 ngmL underwenttransrectal ultrasound-guided prostate biopsy to rule outprostate cancer Prostate size was assessed using transrectalultrasound Patients with prostate cancer neurogenic blad-der urinary tract infection uncontrolled diabetes mellitusor cardiovascular disease were excluded The clinical char-acteristics of BPH patients are summarized in Table 1 Todetermine the relationship between polymorphisms of FGFRgenes and clinical phenotypes BPH patients were dividedinto subgroups according to prostate volume (lt30 mL or ge30mL) PSA level (lt15 or ge15 ngmL) IPSS (lt20 or ge20) and(119876max) (lt10 or ge10mLsec) [20 21]

Normal healthy controls were recruited frommen visitingthe hospital for routine health checkups All healthy controlsubjects underwent screening and had a normal PSA level(lt40 ngmL) They showed no clinical evidence of BPHneurogenic bladder urinary tract infection diabetesmellituscardiovascular disease or any other severe diseases

Written informed consent was obtained from all patientsand control subjects for the use of clinical data and samplesincluding DNA extracted from peripheral bloodThe Institu-tional Review Board at KyungHeeUniversityMedical Centerapproved the protocol for this study

Table 1 Clinical characteristics of benign prostatic hyperplasia(BPH) patients and control subjects

BPH ControlNo of subjects 233 213Age (mean plusmn SD) 6577 plusmn 946 6189 plusmn 826

Prostate volume (mL)Total 3834 plusmn 2138

Inner 1799 plusmn 1860

PSA (ngmL)Total 438 plusmn 524

Free 099 plusmn 122

IPSS 1705 plusmn 766

QoL 361 plusmn 134

Uroflowmetry (mLs)119876max 1133 plusmn 577

119876avg 646 plusmn 367

VV (mL) 10900 plusmn 14833

PVR (mL) 5759 plusmn 9868

PSA prostate-specific antigen IPSS international prostate symptom scoreQoL quality of life119876max peak urinary flow rate119876avg average urinary flowrate VV voided volume PVR postvoid residual urine

22 SNP Selection and Genotyping SNPs located in theexons [51015840-untranslated region (UTR) coding region and31015840UTR] of the FGFR1 and FGFR2 genes were selectedfrom the National Center for Biotechnology InformationSNP database (httpwwwncbinlmnihgovprojectsSNPBUILD 137) We excluded SNPs without data on genotypefrequency and those with a minor allele frequency (MAF)lt 005 in Chinese and Japanese populations Finally weselected four SNPs [rs13317 (TC 31015840UTR) for FGFR1rs755793 (Met186Thr) rs1047100 (Val232Val) and rs3135831(CT 31015840UTR) for FGFR2] DNAwas isolated from peripheralblood samples using a DNA Isolation Kit for blood (RocheIN USA) SNP genotyping was conducted by direct sequen-cing using specific primers for rs13317 (sense 51015840-CCA-ACTTAGTGAAACCCCATCT-31015840 antisense 51015840-CCCAAC-AAATACAGTCTGGTCA-31015840) rs755793 (sense 51015840-TACTCA-TGGAGGGGAAGCTG-31015840 antisense 51015840-CTGACATGG-GCAATTGTGAC-31015840) rs1047100 (sense 51015840-CATACCTTT-CTTGCCTCCTTCA-31015840 antisense 51015840-CAGAAGCAGCCT-TGTAAAATGA-31015840) and rs3135831 (sense 51015840-TGTATTTCC-CAAACCTCTGTCC-31015840 antisense 51015840-CACTGTCAAGGC-TATAAACTGC-31015840) PCR products were sequenced usingan ABI PRISM 3730XL analyzer (PE Applied BiosystemsFoster City CA USA) Sequence data were analyzed usingSeqManII software (DNASTAR Inc Madison WI USA)

23 Statistical Analysis SNPStats (httpbioinfoiconcologianetindexphp) and SPSS 180 software (SPSS Inc ChicagoIL USA) were used to analyze genetic data and deter-mine Hardy-Weinberg equilibrium (HWE) Associa-tions between SNPs and BPH as well as any associationsbetween the SNPs and BPH subgroups were estimated bycomputing odds ratios (ORs) and 95 confidence intervals(CIs) with logistic regression analysis controlling for age

Disease Markers 871

Table 2 Frequencies of the genotypes and alleles of polymorphisms of FGFR genes in BPH patients and control subjects

Gene SNP Genotypeallele Control BPH Models OR (95 CI) 119875119899 = 213 () 119899 = 233 ()

FGFR1 rs13317

TT 85 (399) 83 (356) Additive 111 (083ndash147) 048TC 100 (470) 116 (498) Dominant 116 (079ndash172) 045CC 28 (132) 34 (146) Recessive 109 (063ndash190) 075

T 270 (634) 282 (605) 1C 156 (366) 184 (395) 113 (086ndash148) 038

FGFR2

rs755793

TT 193 (906) 208 (893) Additive 108 (059ndash200) 080TC 20 (94) 24 (103) Dominant 104 (055ndash196) 090CC 0 (00) 1 (04) Recessive NA (000-NA) NA

T 406 (953) 440 (944) 1C 20 (47) 26 (56) 120 (066ndash218) 055

rs1047100

GG 193 (906) 196 (841) Additive 192 (107ndash343) 0024GA 20 (94) 35 (150) Dominant 189 (104ndash345) 0034AA 0 (00) 2 (09) Recessive NA (000-NA) NA

G 406 (953) 427 (916) 1A 20 (47) 39 (84) 185 (106ndash323) 0029

rs3135831

CC 92 (432) 116 (498) Additive 083 (063ndash109) 017CT 88 (413) 90 (386) Dominant 080 (055ndash118) 026TT 33 (155) 27 (116) Recessive 072 (041ndash125) 024

C 272 (638) 322 (691) 1T 154 (362) 144 (309) 079 (060ndash104) 010

as a covariable In the logistic regression analysis for eachSNP the models assuming additive inheritance (the riskincreased r-fold for subjects with one minor allele and2r-fold for subjects with two minor alleles) dominantinheritance (subjects with one or two minor alleles had thesame relative risk for the disease) or recessive inheritance(subjects with two minor alleles were at increased risk ofthe disease) were used To avoid chance findings due tomultiple comparison the Bonferroni correction was appliedby lowering significance levels to 119875 = 0054 for the 4 SNPs

3 Results

All SNPs analyzed in this study were polymorphic and thegenotype distributions of the SNPs were in HWE (119875 gt 005)Differences in genotype distributions and allele frequenciesfor the four SNPs between BPH and control were analyzedAs shown in Table 2 rs1047100 of FGFR2 was associated withBPH in the additive (GAversusAAversusGG119875 = 0024 OR= 192 95 CI = 107ndash343) and dominant models (GAAAversus GG 119875 = 0034 OR = 189 95 CI = 104ndash345)Allele frequency analysis also showed an association betweenrs1047100 and BPH (119875 = 0029 OR = 185 95 CI = 106ndash323) However the statistical significance did not remainafter Bonferroni correction

We further analyzed the associations between SNPs andthe clinical phenotypes of BPH (prostate volume PSA level

IPSS and 119876max) In analysis according to small or largeprostate volume (lt30 or ge30 mL) we found that rs1047100of FGFR2was significantly associatedwith prostate volume inthe additive (GA versus AA versus GG 119875 = 0016 OR = 04395 CI = 021ndash087) and dominant models (GAAA versusGG 119875 = 0010 OR = 038 95 CI = 018ndash081) (Table 3)However statistical significance was only maintained in thedominant model after Bonferroni correction The frequencyof genotypes containing the A allele was lower in BPHpatients with large prostate volume (GA = 96 AA = 08)compared to those with small prostate volume (GA = 215AA = 09) Although allele frequency analysis also revealedthat rs1047100 was associated with prostate volume (119875 =0021 OR = 045 95 CI = 023ndash089) this result was notsignificant after Bonferroni correction

We also found a significant association between rs13317of FGFR1 and IPSS in an analysis according to low or highIPSS (lt20 or ge20) As shown in Table 4 rs13317 of FGFR1wasassociated with IPSS in the additive (TC versus CC versus TT119875 = 00022 OR = 050 95 CI = 032ndash079) and dominantmodels (TCCC versus TT 119875 = 0005 OR = 043 95CI = 023ndash078) Allele frequency analysis also revealed thatrs13317 of FGFR1 was associated with IPSS (119875 = 0005 OR= 055 95 CI = 036ndash084) In particular the frequency ofthe C allele was decreased in BPH patients with high IPSS(303) compared to those with low IPSS (442) Theseresults remained significant after Bonferroni correction

872 Disease Markers

Table 3 Frequencies of the genotypes and alleles of polymorphisms of FGFR genes based on small and large prostate volume in subjectswith BPH

Gene SNP GenotypealleleProstate volume (mL)

Models OR (95 CI) 119875lt30119899 = 107 ()

ge30119899 = 125 ()

FGFR1 rs13317


T 130 (607) 150 (600) 1C 84 (393) 100 (400) 103 (071ndash150) 087

FGFR2

rs755793

TT 93 (869) 114 (912) Additive 054 (024ndash121) 013TC 13 (122) 11 (88) Dominant 055 (023ndash130) 017CC 1 (09) 0 (00) Recessive 000 (000-NA) NA

T 199 (930) 239 (956) 1C 15 (70) 11 (44) 061 (027ndash136) 023

rs1047100

GG 83 (776) 112 (896) Additive 043 (021ndash087) 0016GA 23 (215) 12 (96) Dominant 038 (018ndash081) 0010AA 1 (09) 1 (08) Recessive 070 (004ndash1133) 080

G 189 (883) 236 (944) 1A 25 (117) 14 (56) 045 (023ndash089) 0021

rs3135831


C 144 (673) 176 (704) 1T 70 (327) 74 (296) 087 (058ndash128) 047

Bold characters represent statistically significant values (119875 lt 0054)

In analysis according to other clinical phenotypes (PSAlevel and 119876max) we were not able to find any association ofpolymorphisms in the FGFR genes (data not shown)

4 Discussion

We examined the association of polymorphisms of FGFR1and FGFR2 with BPH and its clinical phenotypes in aKorean population No significant association was detectedbetween polymorphisms of FGFR1 and FGFR2 and BPHHowever in analysis according to clinical phenotypes wefound associations between rs1047100 of FGFR2 and prostatevolume and between rs13317 of FGFR1 and IPSS

BPH is a progressive disease found in many men andnumerous factors including androgen level and aging havebeen linked with the risk of BPH progression [22ndash24]Prostate volume is the most extensively studied risk factorfor BPH progression [25 26] Indeed it was reported thatmen with prostate volume ge 30 mL were more likely to suffermoderate-to-severe symptoms (35-fold increase) decreasedflow rates (25-fold increase) and acute urinary retention (3-to 4-fold increase) compared to men with prostate volumelt 30 mL [27] Reduced urinary flow increased IPSS andincreased PSA have been also suggested as predictors of BPHprogression [28] Clinical study demonstrated that men with

prostate volume ge 31 mL PSA ge 16 ngmL or 119876max lt 106mLsec at baseline had a significantly increased risk of overallclinical progression of BPH [29] Although IPSS and PSAare simple BPH diagnostic factors used in the primary caresetting a previous study showed a high correlation betweenBPH diagnosed by simple tests (medical history IPSS digitalrectal examination (DRE) and PSA) and that diagnosed bya full battery of tests including ultrasonographic assessmentof residual and prostatic volume and uroflowmetry [30]Thus these factors may also be useful as predictors of BPHprogression In our study although polymorphisms of FGFR1and FGFR2 were not associated with PSA level or 119876max inBPH patients rs1047100 of FGFR2 and rs13317 of FGFR1wereassociated with prostate volume and IPSS respectivelyTheseresults indicated that polymorphisms of FGFR1 and FGFR2may be related to the severity of BPH and implicated in theprogression rather than the incidence of BPH In particularwe found that the frequency of genotype containing theminor allele A of rs1047100 in FGFR2 was lower in BPHpatients with large prostate volume than in those with smallprostate volume In addition the frequency of the minorallele C of rs13317 in FGFR1 was significantly decreased inBPH patients with high IPSS These finding indicated thatpatients with genotypes containing the A allele of rs1047100or the C allele of rs13317 may be protected from severeprogression of BPH

Disease Markers 873

Table 4 Frequencies of the genotypes and alleles of polymorphisms of FGFR genes based on low and high international prostate symptomscore (IPSS) in subjects with BPH

Gene SNP GenotypealleleIPSS

Models OR (95 CI) 119875lt20119899 = 130 ()

ge20119899 = 76 ()

FGFR1 rs13317


T 145 (558) 106 (697) 1C 115 (442) 46 (303) 055 (036ndash084) 0005

FGFR2

rs755793


T 242 (931) 146 (961) 1C 18 (69) 6 (39) 055 (021ndash142) 022

rs1047100

GG 107 (823) 67 (882) Additive 060 (027ndash132) 019GA 21 (161) 9 (118) Dominant 063 (027ndash145) 027AA 2 (15) 0 (0) Recessive 000 (000-NA) NAG 235 (904) 143 (941) 1A 25 (96) 9 (59) 059 (027ndash130) 019

rs3135831


C 176 (677) 107 (704) 1T 84 (323) 45 (296) 088 (057ndash136) 057


Previous studies reported that FGFRs were abundantlyexpressed in the normal prostate [11ndash14] and that theexpression of FGFR1 and FGFR2 was elevated in prostatesof BPH patients [16ndash18] To our knowledge there are noreports indicating that the expression of FGFR1 or FGFR2is upregulated or downregulated according to the alleles ofrs1047100 and rs13317 However rs13317 could be involvedin regulating expression considering that it is located in the31015840UTR which modifies stability and transport of mRNA aswell as translation efficiency [31] and whose SNPs are wellknown to increase the efficiency of 31015840 end processing [32]In addition although rs1047100 is synonymous SNP recentstudies reported that synonymous SNPs play an importantrole in protein activities and specificities without influencingamino acid sequences [33 34] Thus we postulated thatFGFR1 and FGFR2 might be overexpressed in BPH but thatthe expression of FGFR1 and FGFR2 might be relatively lowin BPH patients with genotypes containing the A allele ofrs1047100 or the C allele of rs13317 compared to BPH patientswithout those Furthermore a previous study showed thatrs13317 of FGFR1 which plays a role in wound healingand is a positive regulator of skeletal formation [35] wasassociated with delayed bone healing after bone fractureand in particular that the frequency of the C allele ofrs13317 in FGFR1 was increased in individuals with delayed

bone healing compared to those with uneventful healing[36] However in that study no significant associationswere observed between FGF polymorphisms and delayedbone healing [36] Thus they suggested the possibility thatspecific alterations in the receptor despite FGFs functionsmay be involved in triggering the pathologic process duringfracture healing [36] Given this report we also postulatedthat rs13317 may affect the activity of FGFR1 Thus FGFR1activity may be relatively decreased in BPH patients with theC allele of rs13317 resulting in lower and slower progressionof BPH in those individuals Further studies are needed todetermine how FGFR1 and FGFR2 polymorphisms affecttheir expression andor activity in BPH progression

The major limitation of our study was the small samplesize used for comparison within BPH subgroups Howeverthis is the first genetic study on the relationship betweenSNPs of FGFR1 and FGFR2 and BPH Our results revealedassociations between FGFR1 and FGFR2 and the clinicalphenotypes of BPH Further studies with a larger sample sizesare needed to validate our results

5 Conclusions

In conclusion we found that polymorphisms of FGFR1 andFGFR2were not associated with BPHHowever clinical anal-ysis revealed that aFGFR2polymorphismwas associatedwith

874 Disease Markers

prostate volume and a FGFR1 polymorphism was associatedwith IPSS in BPH patients These results suggest that FGFR1and FGFR2may be related to BPH severity and progression

Conflict of Interests

The authors declare that they have no conflict of interests

Acknowledgment

This work was supported by a Grant from Kyung HeeUniversity in 2013 (KHU-20130169)

References

[1] G Kramer D Mitteregger and M Marberger ldquoIs benign pro-static hyperplasia (BPH) an immune inflammatory diseaserdquoEuropean Urology vol 51 no 5 pp 1202ndash1216 2007

[2] G Untergasser S Madersbacher and P Berger ldquoBenign pro-static hyperplasia age-related tissue-remodelingrdquo ExperimentalGerontology vol 40 no 3 pp 121ndash128 2005

[3] NA Bhowmick E GNeilson andH LMoses ldquoStromal fibro-blasts in cancer initiation and progressionrdquoNature vol 432 no7015 pp 332ndash337 2004

[4] K L Lee and DM Peehl ldquoMolecular and cellular pathogenesisof benign prostatic hyperplasiardquo Journal of Urology vol 172 no5 pp 1784ndash1791 2004

[5] DM Peehl and R G Sellers ldquoCultured stromal cells an in vitromodel of prostatic mesenchymal biologyrdquo Prostate vol 45 pp115ndash123 2000

[6] Z Li T Habuchi N Tsuchiya et al ldquoIncreased risk of prostatecancer and benign prostatic hyperplasia associated with trans-forming growth factor-beta 1 gene polymorphism at codon10rdquoCarcinogenesis vol 25 no 2 pp 237ndash240 2004

[7] M D Omrani S Taghipour-Bazargani S Salari-Lak and MBagheri ldquoAssociation of codon 10 polymorphism of the trans-forming growth factor beta 1 gene with prostate cancer andhyperplasia in an iranian populationrdquo Urologia Internationalisvol 83 no 3 pp 329ndash332 2009

[8] R J Mullan E J Bergstralh S A Farmer et al ldquoGrowth factorcytokine and vitamin D receptor polymorphisms and risk ofbenign prostatic hyperplasia in a community-based cohort ofmenrdquo Urology vol 67 no 2 pp 300ndash305 2006

[9] N Tsuchiya L Wang Y Horikawa et al ldquoCA repeat poly-morphism in the insulin-like growth factor-I gene is associatedwith increased risk of prostate cancer and benign prostatichyperplasiardquo International journal of oncology vol 26 no 1 pp225ndash231 2005

[10] R Grose and C Dickson ldquoFibroblast growth factor signaling intumorigenesisrdquoCytokine andGrowth Factor Reviews vol 16 no2 pp 179ndash186 2005

[11] M Ittman andAMansukhani ldquoExpression of fibroblast growthfactors (FGFs) and FGF receptors in human prostaterdquo Journal ofUrology vol 157 no 1 pp 351ndash356 1997

[12] F Sinowatz W Amselgruber D Lincoln et al ldquoRole of basicfibroblast growth factor in prostatic tumorsrdquo Nutrition vol 11no 5 pp 619ndash621 1995

[13] M S Steiner ldquoRole of peptide growth factors in the prostate areviewrdquo Urology vol 42 no 1 pp 99ndash110 1993

[14] C Saez A C Gonzalez-Baena M A Japon et al ldquoExpressionof basic fibroblast growth factor and its receptors FGFR1 andFGFR2 in human benign prostatic hyperplasia treated withfinasteriderdquo Prostate vol 40 pp 83ndash88 1999

[15] F Ropiquet D Giri B Kwabi-addo A Mansukhani and MIttmann ldquoIncreased expression of fibroblast growth factor 6 inhuman prostatic intraepithelial neoplasia and prostate cancerrdquoCancer Research vol 60 no 15 pp 4245ndash4250 2000

[16] S Boget C Cereser P Parvaz A Leriche and A Revol ldquoFibro-blast growth factor receptor 1 (FGFR1) is over-expressed inbenign prostatic hyperplasia whereas FGFR2-IIIc and FGFr3are notrdquo European Journal of Endocrinology vol 145 no 3 pp303ndash310 2001

[17] A Hamaguchi I Tooyama T Yoshiki and H Kimura ldquoDem-onstration of fibroblast growth factor receptor-I in humanprostate by polymerase chain reaction and immunohistochem-istryrdquo Prostate vol 27 no 3 pp 141ndash147 1995

[18] H Y Leung P Mehta L B Gray A T Collins C N Robsonand D E Neal ldquoKeratinocyte growth factor expression inhormone insensitive prostate cancerrdquo Oncogene vol 15 no 9pp 1115ndash1120 1997

[19] ZMa N Tsuchiya T Yuasa et al ldquoPolymorphisms of fibroblastgrowth factor receptor 4 have association with the develop-ment of prostate cancer and benign prostatic hyperplasia andthe progression of prostate cancer in a Japanese populationrdquoInternational Journal of Cancer vol 123 no 11 pp 2574ndash25792008

[20] P Siami CG Roehrborn J Barkin et al ldquoCombination therapywith dutasteride and tamsulosin in men with moderate-to-severe benign prostatic hyperplasia and prostate enlargementthe CombAT (Combination of Avodart and Tamsulosin) trialrationale and study designrdquo Contemporary Clinical Trials vol28 no 6 pp 770ndash779 2007

[21] S A Kaplan J D McConnell C G Roehrborn et al ldquoCombi-nation therapy with doxazosin and finasteride for benign pro-static hyperplasia in patients with lower urinary tract symptomsand a baseline total prostate volume of 25Ml or greaterrdquo Journalof Urology vol 175 no 1 pp 217ndash220 2006 discussion 220-211

[22] C G Roehrborn P Boyle D Bergner et al ldquoSerum prostate-specific antigen and prostate volume predict long-term changesin symptoms and flow rate results of a four-year randomizedtrial comparing finasteride versus placebordquoUrology vol 54 no4 pp 662ndash669 1999

[23] J D McConnell R Bruskewitz P Walsh et al ldquoThe effectof finasteride on the risk of acute urinary retention and theneed for surgical treatment among men with benign prostatichyperplasiardquoTheNew England Journal of Medicine vol 338 no9 pp 557ndash563 1998

[24] S J Jacobsen D J Jacobson C J Girman et al ldquoNatural historyof prostatism risk factors for acute urinary retentionrdquo Journalof Urology vol 158 no 2 pp 481ndash487 1997

[25] H B Shim J K Lee T Y Jung and J H Ku ldquoSerum prostate-specific antigen as a predictor of prostate volume in Koreanmen with lower urinary tract symptomsrdquo Prostate Cancer andProstatic Diseases vol 10 no 2 pp 143ndash148 2007

[26] Y-L ChangA T L Lin K-KChen et al ldquoCorrelation betweenserum prostate specific antigen and prostate volume in Tai-wanese men with biopsy proven benign prostatic hyperplasiardquoJournal of Urology vol 176 no 1 pp 196ndash199 2006

[27] J B Anderson C G Roehrborn J A Schalken and MEmberton ldquoThe progression of benign prostatic hyperplasia

Disease Markers 875

examining the evidence and determining the riskrdquo EuropeanUrology vol 39 no 4 pp 390ndash399 2001

[28] M Emberton E B Cornel P F Bassi R O Fourcade J MF Gomez and R Castro ldquoBenign prostatic hyperplasia as aprogressive disease a guide to the risk factors and options formedicalmanagementrdquo International Journal of Clinical Practicevol 62 no 7 pp 1076ndash1086 2008

[29] E D Crawford S S Wilson J D McConnell et al ldquoBaselinefactors as predictors of clinical progression of benign prostatichyperplasia in men treated with placebordquo Journal of Urologyvol 175 no 4 pp 1422ndash1426 2006

[30] J Carballido Rodrıguez X Badia Llach A Gimeno ColladoL Regadera Sejas R Dal-Re Saavedra and M Guilera SardaldquoValidity of tests for initial diagnosis and its concordance withfinal diagnosis in patients with suspected benign prostatichyperplasiardquo Actas Urologicas Espanolas vol 30 no 7 pp 667ndash674 2006

[31] SDiMarco ZHel C LachanceH Furneaux andDRadziochldquoPolymorphism in the 31015840-untranslated region of TNF120572 mRNAimpairs binding of the post-transcriptional regulatory proteinHuR to TNF120572mRNArdquoNucleic Acids Research vol 29 no 4 pp863ndash871 2001

[32] N H Gehring U Frede G Neu-Yilik et al ldquoIncreasedefficiency ofmRNA31015840 end formation a new geneticmechanismcontributing to hereditary thrombophiliardquoNature Genetics vol28 no 4 pp 389ndash393 2001

[33] A A Komar ldquoSilent SNPs impact on gene function and pheno-typerdquo Pharmacogenomics vol 8 no 8 pp 1075ndash1080 2007

[34] C Kimchi-Sarfaty J M Oh I-W Kim et al ldquoA ldquosilentrdquo poly-morphism in the MDR1 gene changes substrate specificityrdquoScience vol 315 no 5811 pp 525ndash528 2007

[35] K Ishizeki H Saito T Shinagawa N Fujiwara and TNawa ldquoHistochemical and immunohistochemical analysis ofthe mechanism of calcification of Meckelrsquos cartilage duringmandible development in rodentsrdquo Journal of Anatomy vol 194no 2 pp 265ndash277 1999

[36] J M Guimaraes I C Guimaraes M E Duarte et al ldquoPolymor-phisms in BMP4 and FGFR1 genes are associated with fracturenon-unionrdquo Journal of Orthopaedic Research vol 31 no 12 pp1971ndash1979 2013

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014


MEDIATORSINFLAMMATION

of


Behavioural Neurology

EndocrinologyInternational Journal of



Disease Markers


BioMed Research International

OncologyJournal of



Oxidative Medicine and Cellular Longevity


PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of



Computational and Mathematical Methods in Medicine

OphthalmologyJournal of


Diabetes ResearchJournal of



Research and TreatmentAIDS


Gastroenterology Research and Practice


Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

870 Disease Markers

cellular responses by activating a family of four receptortyrosine kinases FGFR1 through FGFR4 [10] FGFRs arepresent in various organs In the humanprostate FGFR1-3 areabundantly expressed in the prostate stroma andor epithe-lium [11ndash14] and FGFR4 has relatively low expression in theepithelium [15] In the prostate FGFRs play an importantrole in prostate organogenesis and perturbations in FGFRexpression have been potently implicated in prostatic disease[11] Previous studies have reported that overexpression ofFGFRs is important in the development of BPH [16ndash18]Indeed FGFR1 was observed to be increased in the stromaof BPH patients compared to that from normal prostates[16 17] and increased expression of FGFR2 has been detectedin BPH [18] Furthermore although no studies have reportedchanges in expression of FGFR4 in BPH a genetic studyshowed a significant association between FGFR4 polymor-phisms and BPH in a Japanese population [19] In light ofthese findings we postulated that FGFRs may be involvedin the pathogenesis of BPH In this study we investigatedthe genetic associations between BPH and the FGFR genes(FGFR1 and FGFR2) in Korean BPH patients by analyz-ing single nucleotide polymorphisms (SNPs) of the FGFRgenes

2 Methods

21 Subjects A total of 233 male patients with BPH (meanage plusmn standard deviation 6577 plusmn 946 years) and 213 malecontrol subjects (mean age 6189 plusmn 826 years) were enrolledAll patients with BPH were from Kyung Hee UniversityHospital between January 2002 and December 2008 and inall patients lower urinary tract symptoms were quantifiedusing IPSS Uroflowmetry was performed to measure peakurinary flow rate (119876max) for all patients Serum prostate-specific antigen (PSA) level wasmeasured in all BPHpatientsPatients with serumPSA level more than 4 ngmL underwenttransrectal ultrasound-guided prostate biopsy to rule outprostate cancer Prostate size was assessed using transrectalultrasound Patients with prostate cancer neurogenic blad-der urinary tract infection uncontrolled diabetes mellitusor cardiovascular disease were excluded The clinical char-acteristics of BPH patients are summarized in Table 1 Todetermine the relationship between polymorphisms of FGFRgenes and clinical phenotypes BPH patients were dividedinto subgroups according to prostate volume (lt30 mL or ge30mL) PSA level (lt15 or ge15 ngmL) IPSS (lt20 or ge20) and(119876max) (lt10 or ge10mLsec) [20 21]

Normal healthy controls were recruited frommen visitingthe hospital for routine health checkups All healthy controlsubjects underwent screening and had a normal PSA level(lt40 ngmL) They showed no clinical evidence of BPHneurogenic bladder urinary tract infection diabetesmellituscardiovascular disease or any other severe diseases

Written informed consent was obtained from all patientsand control subjects for the use of clinical data and samplesincluding DNA extracted from peripheral bloodThe Institu-tional Review Board at KyungHeeUniversityMedical Centerapproved the protocol for this study

Table 1 Clinical characteristics of benign prostatic hyperplasia(BPH) patients and control subjects

BPH ControlNo of subjects 233 213Age (mean plusmn SD) 6577 plusmn 946 6189 plusmn 826

Prostate volume (mL)Total 3834 plusmn 2138

Inner 1799 plusmn 1860

PSA (ngmL)Total 438 plusmn 524

Free 099 plusmn 122

IPSS 1705 plusmn 766

QoL 361 plusmn 134

Uroflowmetry (mLs)119876max 1133 plusmn 577

119876avg 646 plusmn 367

VV (mL) 10900 plusmn 14833

PVR (mL) 5759 plusmn 9868

PSA prostate-specific antigen IPSS international prostate symptom scoreQoL quality of life119876max peak urinary flow rate119876avg average urinary flowrate VV voided volume PVR postvoid residual urine

22 SNP Selection and Genotyping SNPs located in theexons [51015840-untranslated region (UTR) coding region and31015840UTR] of the FGFR1 and FGFR2 genes were selectedfrom the National Center for Biotechnology InformationSNP database (httpwwwncbinlmnihgovprojectsSNPBUILD 137) We excluded SNPs without data on genotypefrequency and those with a minor allele frequency (MAF)lt 005 in Chinese and Japanese populations Finally weselected four SNPs [rs13317 (TC 31015840UTR) for FGFR1rs755793 (Met186Thr) rs1047100 (Val232Val) and rs3135831(CT 31015840UTR) for FGFR2] DNAwas isolated from peripheralblood samples using a DNA Isolation Kit for blood (RocheIN USA) SNP genotyping was conducted by direct sequen-cing using specific primers for rs13317 (sense 51015840-CCA-ACTTAGTGAAACCCCATCT-31015840 antisense 51015840-CCCAAC-AAATACAGTCTGGTCA-31015840) rs755793 (sense 51015840-TACTCA-TGGAGGGGAAGCTG-31015840 antisense 51015840-CTGACATGG-GCAATTGTGAC-31015840) rs1047100 (sense 51015840-CATACCTTT-CTTGCCTCCTTCA-31015840 antisense 51015840-CAGAAGCAGCCT-TGTAAAATGA-31015840) and rs3135831 (sense 51015840-TGTATTTCC-CAAACCTCTGTCC-31015840 antisense 51015840-CACTGTCAAGGC-TATAAACTGC-31015840) PCR products were sequenced usingan ABI PRISM 3730XL analyzer (PE Applied BiosystemsFoster City CA USA) Sequence data were analyzed usingSeqManII software (DNASTAR Inc Madison WI USA)

23 Statistical Analysis SNPStats (httpbioinfoiconcologianetindexphp) and SPSS 180 software (SPSS Inc ChicagoIL USA) were used to analyze genetic data and deter-mine Hardy-Weinberg equilibrium (HWE) Associa-tions between SNPs and BPH as well as any associationsbetween the SNPs and BPH subgroups were estimated bycomputing odds ratios (ORs) and 95 confidence intervals(CIs) with logistic regression analysis controlling for age

Disease Markers 871



FGFR1 rs13317


T 270 (634) 282 (605) 1C 156 (366) 184 (395) 113 (086ndash148) 038

FGFR2

rs755793


T 406 (953) 440 (944) 1C 20 (47) 26 (56) 120 (066ndash218) 055

rs1047100


G 406 (953) 427 (916) 1A 20 (47) 39 (84) 185 (106ndash323) 0029

rs3135831


C 272 (638) 322 (691) 1T 154 (362) 144 (309) 079 (060ndash104) 010


3 Results





872 Disease Markers



Models OR (95 CI) 119875lt30119899 = 107 ()

ge30119899 = 125 ()

FGFR1 rs13317


T 130 (607) 150 (600) 1C 84 (393) 100 (400) 103 (071ndash150) 087

FGFR2

rs755793


T 199 (930) 239 (956) 1C 15 (70) 11 (44) 061 (027ndash136) 023

rs1047100


G 189 (883) 236 (944) 1A 25 (117) 14 (56) 045 (023ndash089) 0021

rs3135831


C 144 (673) 176 (704) 1T 70 (327) 74 (296) 087 (058ndash128) 047



4 Discussion




Disease Markers 873



Models OR (95 CI) 119875lt20119899 = 130 ()

ge20119899 = 76 ()

FGFR1 rs13317


T 145 (558) 106 (697) 1C 115 (442) 46 (303) 055 (036ndash084) 0005

FGFR2

rs755793


T 242 (931) 146 (961) 1C 18 (69) 6 (39) 055 (021ndash142) 022

rs1047100


rs3135831


C 176 (677) 107 (704) 1T 84 (323) 45 (296) 088 (057ndash136) 057





5 Conclusions


874 Disease Markers




Acknowledgment


References




























Disease Markers 875
















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















Disease Markers 871



FGFR1 rs13317


T 270 (634) 282 (605) 1C 156 (366) 184 (395) 113 (086ndash148) 038

FGFR2

rs755793


T 406 (953) 440 (944) 1C 20 (47) 26 (56) 120 (066ndash218) 055

rs1047100


G 406 (953) 427 (916) 1A 20 (47) 39 (84) 185 (106ndash323) 0029

rs3135831


C 272 (638) 322 (691) 1T 154 (362) 144 (309) 079 (060ndash104) 010


3 Results





872 Disease Markers



Models OR (95 CI) 119875lt30119899 = 107 ()

ge30119899 = 125 ()

FGFR1 rs13317


T 130 (607) 150 (600) 1C 84 (393) 100 (400) 103 (071ndash150) 087

FGFR2

rs755793


T 199 (930) 239 (956) 1C 15 (70) 11 (44) 061 (027ndash136) 023

rs1047100


G 189 (883) 236 (944) 1A 25 (117) 14 (56) 045 (023ndash089) 0021

rs3135831


C 144 (673) 176 (704) 1T 70 (327) 74 (296) 087 (058ndash128) 047



4 Discussion




Disease Markers 873



Models OR (95 CI) 119875lt20119899 = 130 ()

ge20119899 = 76 ()

FGFR1 rs13317


T 145 (558) 106 (697) 1C 115 (442) 46 (303) 055 (036ndash084) 0005

FGFR2

rs755793


T 242 (931) 146 (961) 1C 18 (69) 6 (39) 055 (021ndash142) 022

rs1047100


rs3135831


C 176 (677) 107 (704) 1T 84 (323) 45 (296) 088 (057ndash136) 057





5 Conclusions


874 Disease Markers




Acknowledgment


References




























Disease Markers 875
















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















872 Disease Markers



Models OR (95 CI) 119875lt30119899 = 107 ()

ge30119899 = 125 ()

FGFR1 rs13317


T 130 (607) 150 (600) 1C 84 (393) 100 (400) 103 (071ndash150) 087

FGFR2

rs755793


T 199 (930) 239 (956) 1C 15 (70) 11 (44) 061 (027ndash136) 023

rs1047100


G 189 (883) 236 (944) 1A 25 (117) 14 (56) 045 (023ndash089) 0021

rs3135831


C 144 (673) 176 (704) 1T 70 (327) 74 (296) 087 (058ndash128) 047



4 Discussion




Disease Markers 873



Models OR (95 CI) 119875lt20119899 = 130 ()

ge20119899 = 76 ()

FGFR1 rs13317


T 145 (558) 106 (697) 1C 115 (442) 46 (303) 055 (036ndash084) 0005

FGFR2

rs755793


T 242 (931) 146 (961) 1C 18 (69) 6 (39) 055 (021ndash142) 022

rs1047100


rs3135831


C 176 (677) 107 (704) 1T 84 (323) 45 (296) 088 (057ndash136) 057





5 Conclusions


874 Disease Markers




Acknowledgment


References




























Disease Markers 875
















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















Disease Markers 873



Models OR (95 CI) 119875lt20119899 = 130 ()

ge20119899 = 76 ()

FGFR1 rs13317


T 145 (558) 106 (697) 1C 115 (442) 46 (303) 055 (036ndash084) 0005

FGFR2

rs755793


T 242 (931) 146 (961) 1C 18 (69) 6 (39) 055 (021ndash142) 022

rs1047100


rs3135831


C 176 (677) 107 (704) 1T 84 (323) 45 (296) 088 (057ndash136) 057





5 Conclusions


874 Disease Markers




Acknowledgment


References




























Disease Markers 875
















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















874 Disease Markers




Acknowledgment


References




























Disease Markers 875
















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















Disease Markers 875
















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















Research Article Involvement of Fibroblast Growth Factor...

Documents

Transcript of Research Article Involvement of Fibroblast Growth Factor...