1

FGFR2 Genetic Polymorphisms Modify the Association of Oral Contraceptive Use with the Risk of Breast Cancer in

Chinese Women

徐 望 红wanghong.xu@fudan.edu.cn

复旦大学公共卫生学院流行病学教研室上海市肿瘤研究所流行病室

研 究 背 景• Breast cancer

– most common cancer in women

– a hormone-related malignancy

– a complex and multi-factorial disease

– a result of interplays between different exposures and host susceptibility

• Oral contraceptives (OC)– exogenous hormones – classified as group 1 carcinogens Schneider HP, et al. Climacteric. 2005;8:311-6. Cogliano V, et al. Lancet Oncol. 2005;6:552-3.

– consistent positive association with breast cancer risk in western populations

Kahlenborn C, et al. Mayo Clin Proc. 2006;81(10):1290-302. Collaborative Group on Hormonal Factors in Breast Cancer. Lancet. 1996;347:1713-27.

– null association in Asian populations Kawai M, et al.Cancer Causes Control. 2010;21(1):135-45. Dorjgochoo T, et al. Int J Cancer. 2009;124(10):2442-9. Ursin G, et al. Am J Epidemiol. 1999;150(6):561-7. Huang CS, et al. Br J Cancer. 1999;80(11):1838-43. Yuan JM, et al. Cancer Res. 1988;48(7):1949-53.

• FGFR2 gene – well recognized breast cancer susceptibility gene

– multiple SNPs in association with breast cancer risk across ethnic groups

– a sex hormone-dependent role of FGFR2 gene in breast cancer

FGFR2 genetic polymorphisms may modify the

association between OC use and breast cancer

risk in Chinese women

研 究 假 设

材 料 与 方 法A two-stage population-based case-control study

SBCS-I (1996-1998)1459 of 1602 newly-diagnosed breast cancer cases at age 25-65 1556 of 1724 eligible controls 1193 (82%) cases and 1310 (84%) controls donated a blood sample

SBCS-II (2002-2005)1989 incident cases (83.7%) aged 25 to 70 years old1989 population controls (70.4%)969 (48.7%) cases and 975 (49.0%) controls provided a blood sample

Epidemiological dataDemographic characteristicsMenstrual and reproductive historyOCs use and postmenopausal HRTPrior disease historyPhysical activityTobacco and alcohol use Diet Weight historyFamily history of cancerMeasured body weight, height, and circumferences of the

waist and hips

Genotyping dataFrom a GWAS scan covering about 120 kb of the FGFR2

gene plus 10 kb region of its 5' upstream and 5 kb region of

its 3' downstream

Genotyping conducted using Affymetrix Genome-Wide

Human SNP array 6.0 following Affymetrix’s protocol 42 SNPs genotyped 19 had minor allele frequency of > 0.05

Imputation conducted using the program MACH 93 imputed SNPs 26 SNPs had minor allele frequency of > 0.05 and average imputation

quality score of > 0.90.

结 果

CharacteristicsStage I Study Stage II Study

Cases(n=1104)

Controls(n=1109)

P valueCases

(n=969 )Controls(n=975)

P value

Age (yrs), mean ± SD 47.6 ± 8.0 47.7 ± 8.3 0.75 51.4 ± 8.3 51.4 ± 8.3 0.91

Education, %

    Elementary school or below 12.4 15.2 7.1 11.8

    Middle school 44.3 42.7 42.9 42.0

High school 31.3 31.9 37.7 36.9

    College or above 12.1 10.3 0.17 12.3 9.3 0.0016

Breast cancer in 1st degree relatives, % 3.4 2.6 0.31 5.7 3.4 0.02

Ever had breast fibroadenoma, % 9.5 4.6 <0.0001 10.0 6.5 0.0051

Age at menarche (yrs.), mean ± SD 14.5 ± 1.6 14.8 ± 1.7 0.0002 14.5 ± 1.7 14.7 ± 1.8 0.0037

Ever had a live birth, % 95.1 96.2 0.20 95.2 96.1 0.30

Number of live births, mean ± SD 1.5 ± 0.8 1.5 ± 0.9 0.15 1.3 ± 0.7 1.4 ± 0.8 0.005

    Age at first live birth (yrs), mean ± SD 26.8 ± 4.1 26.3 ± 3.8 0.003 26.1 ± 3.7 25.7 ± 3.8 0.03

Post-menopausal, % 32.8 36.6 0.06 45.5 47.1 0.49

    Age at menopause (yrs.), mean ± SD 48.1 ± 4.7 47.4 ± 5.0 0.04 48.7 ± 4.4 48.0 ± 4.5 0.03

Oral contraceptive use, % 22.1 22.2 0.95 18.1 18.5 0.82

Duration of OC use, mean ± SD 8.6 ± 31.4 9.4 ± 32.9 0.52 8.3 ± 31.6 7.5 ± 28.1 0.56

HRT use among postmenopausal women, % 3.1 5.0 0.18 8.4 4.6 0.02

Physical active in past 10 years, % 19.2 26.3 <0.0001 31.8 34.7 0.18

BMI (kg/m2), mean ± SD 23.6 ± 3.4 23.3 ± 3.4 0.02 24.0 ± 3.3 23.4 ± 3.3 <0.0001

a From χ2 test (categorical variables) or t test (continuous variables).

Table 1. Comparison of demographic characteristics between breast cancer

cases and controls, the Shanghai Breast Cancer Study

Sex hormones use Cases/Controls Adjusted OR (95%CI)Oral contraceptives use

Never 1654/1657 1.00Ever 419/426 0.98(0.83-1.14)

Type of OC usedNever 1654/1657 1.00Progesterone plus estrogen 328/324 1.06(0389-1.26)Progesterone only 42/52 0.86(0.57-1.31)Estrogen only 24/30 0.80(0.46-1.38)Others 25/20 1.30(0.72-2.37)

Duration of OC use Never 1654/1657 1.00

<18 months 208/225 0.95(0.77-1.16) ≥ 18 months 211/201 1.12(0.90-1.38)

P for trend=0.48HRT use (postmenopausal women only)

Never 753/822 1.00Ever 48/41 1.14(0.74-1.77)

Table 2. Association of sex hormones use and breast cancer risk among Chinese women, the Shanghai Breast Cancer Study

OR: adjusted for age, educational levels, age at menarche, menopausal status, breast cancer in 1st-degree relatives, number of live births and body mass index.

All subjects Non-OC users OC-users P for interaction

rs10749418

rs1649161

rs1649204

rs1047057

rs1649202

rs2556537

rs2278202

rs7090018

rs2912759

rs11199991

rs2912760

rs3750813

rs2981457

rs7073360

rs3135749

rs3135747

rs755793

rs3135739

rs3135737

rs3135736

rs12245334

rs9888022

rs2303568

rs2912787

rs2981449

rs3135730

rs2981432

rs10736303

rs9787582

rs11200014

rs1078806

rs2981575

rs1219648

rs1219643

rs2912774

rs2936870

rs17102287

rs2860197

rs2420946

rs2162540

rs2981582

rs2981581

rs3135715

rs7899765

rs1219627

ID

Study

1.4 .6 .8 1 1.21.41.61.8 2 2.22.42.62.8 3 3.23.4

All subject

rs10749418

rs1649161

rs1649204

rs1047057

rs1649202

rs2556537

rs2278202

rs7090018

rs2912759

rs11199991

rs2912760

rs3750813

rs2981457

rs7073360

rs3135749

rs3135747

rs755793

rs3135739

rs3135737

rs3135736

rs12245334

rs9888022

rs2303568

rs2912787

rs2981449

rs3135730

rs2981432

rs10736303

rs9787582

rs11200014

rs1078806

rs2981575

rs1219648

rs1219643

rs2912774

rs2936870

rs17102287

rs2860197

rs2420946

rs2162540

rs2981582

rs2981581

rs3135715

rs7899765

rs1219627

ID

Study

.58

.05

.05

.05

.31

.05

.16

.59

.49

.19

.09

.19

.77

.02

.02

.02

.02

.03

.03

.03

.03

.03

.01

.21

.67

.03

.97

.15

.11

.07

.02

.2

.57

.14

.49

.34

.08

.38

.37

.46

.07

.31

.35

.3

.13

p

.58

.05

.05

.05

.31

.05

.16

.59

.49

.19

.09

.19

.77

.02

.02

.02

.02

.03

.03

.03

.03

.03

.01

.21

.67

.03

.97

.15

.11

.07

.02

.2

.57

.14

.49

.34

.08

.38

.37

.46

.07

.31

.35

.3

.13

p

1.4 .6 .8 1 1.21.41.61.8 2 2.22.42.62.8 3 3.23.4

Ever

rs10749418

rs1649161

rs1649204

rs1047057

rs1649202

rs2556537

rs2278202

rs7090018

rs2912759

rs11199991

rs2912760

rs3750813

rs2981457

rs7073360

rs3135749

rs3135747

rs755793

rs3135739

rs3135737

rs3135736

rs12245334

rs9888022

rs2303568

rs2912787

rs2981449

rs3135730

rs2981432

rs10736303

rs9787582

rs11200014

rs1078806

rs2981575

rs1219648

rs1219643

rs2912774

rs2936870

rs17102287

rs2860197

rs2420946

rs2162540

rs2981582

rs2981581

rs3135715

rs7899765

rs1219627

ID

Study

1.4 .6 .8 1 1.21.41.61.8 2 2.22.42.62.8 3 3.23.4

All subject

rs10749418

rs1649161

rs1649204

rs1047057

rs1649202

rs2556537

rs2278202

rs7090018

rs2912759

rs11199991

rs2912760

rs3750813

rs2981457

rs7073360

rs3135749

rs3135747

rs755793

rs3135739

rs3135737

rs3135736

rs12245334

rs9888022

rs2303568

rs2912787

rs2981449

rs3135730

rs2981432

rs10736303

rs9787582

rs11200014

rs1078806

rs2981575

rs1219648

rs1219643

rs2912774

rs2936870

rs17102287

rs2860197

rs2420946

rs2162540

rs2981582

rs2981581

rs3135715

rs7899765

rs1219627

ID

Study

1.4 .6 .8 1 1.21.41.61.8 2 2.22.42.62.8 3 3.23.4

Never

rs1 0 7 4 9 4 1 8

rs1 6 4 9 1 6 1

rs1 6 4 9 2 0 4

rs1 0 4 7 0 5 7

rs1 6 4 9 2 0 2

rs2 5 5 6 5 3 7

rs2 2 7 8 2 0 2

rs7 0 9 0 0 1 8

rs2 9 1 2 7 5 9

rs1 1 1 9 9 9 9 1

rs2 9 1 2 7 6 0

rs3 7 5 0 8 1 3

rs2 9 8 1 4 5 7

rs7 0 7 3 3 6 0

rs3 1 3 5 7 4 9

rs3 1 3 5 7 4 7

rs7 5 5 7 9 3

rs3 1 3 5 7 3 9

rs3 1 3 5 7 3 7

rs3 1 3 5 7 3 6

rs1 2 2 4 5 3 3 4

rs9 8 8 8 0 2 2

rs2 3 0 3 5 6 8

rs2 9 1 2 7 8 7

rs2 9 8 1 4 4 9

rs3 1 3 5 7 3 0

rs2 9 8 1 4 3 2

rs1 0 7 3 6 3 0 3

rs9 7 8 7 5 8 2

rs1 1 2 0 0 0 1 4

rs1 0 7 8 8 0 6

rs2 9 8 1 5 7 5

rs1 2 1 9 6 4 8

rs1 2 1 9 6 4 3

rs2 9 1 2 7 7 4

rs2 9 3 6 8 7 0

rs1 7 1 0 2 2 8 7

rs2 8 6 0 1 9 7

rs2 4 2 0 9 4 6

rs2 1 6 2 5 4 0

rs2 9 8 1 5 8 2

rs2 9 8 1 5 8 1

rs3 1 3 5 7 1 5

rs7 8 9 9 7 6 5

rs1 2 1 9 6 2 7

ID

S tu d y

1.4 .6 .8 1 1 .2 1 .4 1 .6 1 .8 2 2 .2 2 .4 2 .6 2 .8 3 3 .2 3 .4

A ll su b je c t

Figure 1. Age-adjusted ORs and 95%CI for breast cancer with FGFR2 genetic polymorphisms by OC use, the Shanghai Breast Cancer Study

OR per allele: adjusted for age, educational levels, age at menarche, menopausal status, breast cancer in 1st-degree relatives, number of live births and body mass index

OC use

No. of MA

OC use

OC use OC use

No. of MA

No. of MA

No. of MA

rs2303568

rs3135730rs1078806

rs755793

Figure 2. Breast cancer odds ratios estimates for OC use according to the number of minor alleles of FGFR2 SNPs, the Shanghai Breast Cancer Study

Hap1: in the order of rs7073360, rs3135749, rs3135747, rs755793, rs3135739, rs3135737, rs3135736, rs12245334 and rs9888022. Hap2: in the order of rs2303568 and rs3135730.Hap3: in the order of rs7073360, rs3135749, rs3135747, rs755793, rs3135739, rs3135737, rs3135736, rs12245334, rs9888022, rs2303568, rs3135730 and rs1078806.OR: adjusted for age, educational levels, age at menarche, menopausal status, breast cancer in 1st-degree relatives, number of live births and body mass index.a additionally adjusted for oral contraceptive use.

Table 3. Haplotype analysis of FGFR2 with the risk of breast cancer under dominant model, the Shanghai Breast Cancer Study

All subjects a Non-OC users OC users Using OC<18 months Using OC ≥18 months

% OR(95%CI) P-value OR(95%CI) P-value OR(95%CI) P-value OR(95%CI) P-value OR(95%CI) P-value

Hap1

TAAACAATT 93.8 1.00 1.00 1.00 1.00 1.00

GGGGTGCCC 6.2 1.25(1.05-1.49) 0.01 1.12(0.92-1.36) 0.27 1.95(1.30-2.92) 0.0012 1.61(0.92-2.81) 0.09 2.41(1.33-4.37) 0.0004

Hap2

AA 92.5 1.00 1.00 1.00 1.00 1.00

GG 6.5 1.22(1.02-1.46) 0.03 1.08(0.88-1.32) 0.47 2.02(1.34-3.04) 0.0008 1.79(1.01-3.17) 0.05 2.29(1.27-4.15) 0.0018

Hap3

TAAACAATTAAA 62.9 1.00 1.00 1.00 1.00 1.00

TAAACAATTAAG 28.4 1.13(1.00-1.27) 0.04 1.08(0.95-1.23) 0.26 1.36(1.05-1.77) 0.02 1.20(0.84-1.72) 0.32 1.60(1.11-2.31)

GGGGTGCCCGGA 4.2 1.31(1.05-1.64) 0.02 1.13(0.88-1.45) 0.33 2.36(1.42-3.92) 0.0009 2.85(1.39-5.85) 0.004 2.34(1.15-4.76) <0.0001

讨 论• First study reporting the modifying effect of FGFR2 in the OCs-

breast cancer association

– effect of OC use in breast cancer depends on the genotypes of FGFR2 at

rs755793, rs2303568, 3135730 and rs1078806

– dose-response relationship between duration of OC use and cancer risk

among women carrying at least one minor allele at rs755793, rs2303568

and rs3135730 or those having two minor alleles at rs1078806

– unclear function of the loci and poorly understood mechanism underlying

the results

•Strengths

– population-based study design

– large sample size

– high participation rate

– homogeneous ethnic background

– extensive coverage of the FGFR2 gene

Limitations

possibility of chance

very few users of OCs containing estrogen or progesterone only

no enough statistical power to evaluate the interaction between

HRT use and FGFR2

Significance

test our hypothesis

provide a possible explain for the null OC-breast cancer

association in Chinese women

support the hormone-dependent nature of FGFR2 gene

important implications in personalized prevention of breast

cancer

致 谢美国 Vanderbilt 大学 上海市肿瘤研究所

• 高玉堂• 项永兵

• Wei Zheng• Xiao-ou Shu • Jirong Long• Qiuyin Cai• Qi Dai

上海市疾病预防控制中心• 卢伟• 郑莹• 顾凯• 鲍萍萍

复旦大学公共卫生学院• 赵根明

This research was supported by NSFC 30872180