Biomarkers
ISSN: 1354-750X (Print) 1366-5804 (Online) Journal homepage: http://www.tandfonline.com/loi/ibmk20
Interleukin 17A rs3819024 A>G polymorphism is associated with an increased risk of gastric cardia adenocarcinoma in a Chinese population Zhengbing Ren, Mingna Li, Ruiping Liu, Yong Wang & Haiyong Gu To cite this article: Zhengbing Ren, Mingna Li, Ruiping Liu, Yong Wang & Haiyong Gu (2014) Interleukin 17A rs3819024 A>G polymorphism is associated with an increased risk of gastric cardia adenocarcinoma in a Chinese population, Biomarkers, 19:5, 411-416 To link to this article: http://dx.doi.org/10.3109/1354750X.2014.924158
Published online: 04 Jun 2014.
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Date: 12 September 2015, At: 13:35
http://informahealthcare.com/bmk ISSN: 1354-750X (print), 1366-5804 (electronic) Biomarkers, 2014; 19(5): 411–416 ! 2014 Informa UK Ltd. DOI: 10.3109/1354750X.2014.924158
RESEARCH ARTICLE
Interleukin 17A rs3819024 A 4 G polymorphism is associated with an increased risk of gastric cardia adenocarcinoma in a Chinese population Zhengbing Ren1*, Mingna Li2*, Ruiping Liu3, Yong Wang4#, and Haiyong Gu1# Department of Cardiothoracic Surgery, Affiliated People’s Hospital of Jiangsu University, Zhenjiang, China, 2Department of Pathology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China, 3Department of Orthopedics, Affiliated Hospital of Nanjing Medical University, Changzhou Second People’s Hospital, Changzhou, China, and 4Department of Cardiothoracic Surgery, Affiliated Hospital of Nanjing Medical University, Changzhou Second People’s Hospital, Changzhou, China
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1
Abstract
Keywords
Gastric cardia adenocarcinoma (GCA) is one of the most common malignant tumors. In addition to environmental risk factors, genetic factors might play an important role in GCA carcinogenesis. To evaluate the association between polymorphisms in the interleukin 17A (IL17A) gene on the development of GCA, we conducted a hospital-based case–control study. A total of 243 GCA cases and 476 controls were recruited and their genotypes were determined using a custom-by-design 48-Plex SNPscanä Kit. IL17A rs3819024 A4G polymorphism was found to be associated with the increased risk of GCA. When the IL17A rs3819024 AA homozygote genotype was used as the reference group, the AG genotype was associated with a significantly increased risk of GCA (AG versus AA: adjusted OR ¼ 1.53, 95% CI ¼ 1.05–2.23, p ¼ 0.026). However, there was no significant association between five other SNPs and GCA. Stratified analyses indicated that a significantly increased risk of GCA associated with the IL17A rs3819024 A4G polymorphism was evident among male patients, patients who drank alcohol or those who never smoked. These findings indicated that functional polymorphism IL17A rs3819024 A4G might contribute to GCA susceptibility. Future larger studies with more rigorous study designs are required to confirm the current findings.
Gastric cardia adenocarcinoma, IL17A, molecular epidemiology, polymorphisms
Introduction Gastric cardia adenocarcinoma (GCA) is one of the most common malignant tumors worldwide (Zhang et al., 2010). GCA has a multi-factorial etiology: both genetic and environmental factors are suspected to contribute to GCA development. Besides smoking and heavy drinking, single nucleotide polymorphisms (SNPs) might partly explain individual differences in GCA susceptibility (Zhang et al., 2012). Infection and chronic inflammation contribute to about 1/4 of all cancer cases (Hussain & Harris, 2007). Cytokines are key regulators to promote or restrain inflammation, and therefore, can either stimulate or inhibit tumor growth and progression (Lin & Karin, 2007; Smyth et al., 2004).
*Zhengbing Ren and Mingna Li contributed equally to this work. # Yong Wang and Haiyong Gu are responsible for statistical design/ analysis. E-mail: [email protected]; haiyong_gu@hotmail. com. Address for correspondence: Professor Yong Wang, Department of Cardiothoracic Surgery, Changzhou Second People’s Hospital, Affiliated Hospital of Nanjing Medical University, Changzhou 213003, China. E-mail: [email protected] Dr. Haiyong Gu, Department of Cardiothoracic Surgery, Affiliated People’s Hospital of Jiangsu University, Zhenjiang 212000, China. E-mail: [email protected]
History Received 31 March 2014 Revised 9 May 2014 Accepted 11 May 2014 Published online 4 June 2014
Interleukin 17A (IL17A), also known as IL17, is the signature T helper 17 (Th17) cell effector cytokine (Korn et al., 2009). IL17A plays an important role in inflammation and leads to the induction of many proinflammatory factors, including tumor necrosis factor-alpha (TNF-a), IL6 and IL1b (Ruddy et al., 2004). High expression of IL17A was observed in various tumor tissues, including multiple myeloma, gastric cancer and breast cancer (Alexandrakis et al., 2006; Zhang et al., 2008; Zhu et al., 2008). Through stimulation of vascular endothelial cell migration and regulation of a series of proangiogenic factors, IL17A enhances angiogenesis (Numasaki et al., 2003). IL17A also mediates tissue remodeling by stimulating the production of angiogenic factors and matrix metalloproteases (Jovanovic et al., 2000). IL17A contributes to the development and progression of prostate cancer, gastric carcinomas and skin cancer (Chen et al., 2011; He et al., 2012; Steiner et al., 2003). IL17A polymorphisms have been associated with the risk of various types of cancer, including esophageal cancer, gastric cancer, breast cancer and cervical cancer (Arisawa et al., 2012; Quan et al., 2012; Wang et al., 2012a; Yin et al., 2014). The IL17A gene is located on human chromosome 6p12.1. The wellstudied IL17A rs2275913 G4A polymorphism is located within a binding motif for nuclear factor activated T cells and
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plays a functional role in the promoter activity of the IL17A gene (Liu et al., 2004). We previously conducted a hospital-based case–control study to evaluate IL17A rs2275913 G4A, rs3819024 A4G, rs3819025 G4A, rs4711998 A4G, rs8193036 C4T and rs8193037 G4A polymorphisms and esophageal squamous cell carcinoma (ESCC) risk (Yin et al., 2014). We found that the IL17A rs4711998 A4G AG genotype was associated with a significantly decreased risk of ESCC (Yin et al., 2014). In this investigation, we evaluated the association between the six functional SNPs and GCA susceptibility in 243 GCA cases and 476 controls.
Patients and methods
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Ethical approval of the study protocol This hospital-based case–control study was approved by the Review Board of Jiangsu University (Zhenjiang, China). We have complied with the World Medical Association Declaration of Helsinki regarding ethical conduct of research involving human subjects and/or animals. All subjects provided written informed consent to be included in the study. Study subjects Two-hundred and forty-three subjects with gastric cardia cancer were consecutively recruited from the Affiliated People’s Hospital of Jiangsu University and Affiliated Hospital of Jiangsu University (Zhenjiang, China) between October 2008 and July 2010. All cases of gastric cardia cancer were diagnosed as GCA by pathologic means. The exclusion criteria were patients who previously had: cancer; any metastasized cancer; radiotherapy or chemotherapy. Among 476 controls, 380 controls were patients without cancer frequency-matched to the cases with regard to age (±5 years) and sex recruited from the two hospitals mentioned above during the same time period. Another 96 controls were recruited from hospitals in Changzhou city (which is a neighboring city of Zhenjiang) as previous described (Cheng et al., 2012). Most of the controls were admitted to the hospitals for the treatment of trauma. Each subject was personally questioned by trained interviewers using a pre-tested questionnaire to obtain information on demographic data (e.g. age, sex) and related risk factors (including tobacco smoking and alcohol consumption). After the interview, 2-mL samples of venous blood were collected from each subject. Individuals who smoked one cigarette per day for 41 year were defined as ‘‘smokers’’. Subjects who consumed 3 alcoholic drinks a week for 46 months were considered to be ‘‘alcohol drinkers’’. Tumor, nodal, and metastatic (TNM) stage was according to the seventh edition of the UICC TNM staging (Kwon, 2011). Isolation of DNA, SNPs selection and genotyping by a custom-by-design 48-Plex SNPscan Kit Blood samples were collected from patients using Vacutainers and transferred to tubes lined with ethylenediamine tetraacetic acid (EDTA). Genomic DNA was isolated from whole blood with the QIAamp DNA Blood Mini Kit (Qiagen, Hilden, Germany) (Zheng et al., 2013). The IL17A SNPs
Biomarkers, 2014; 19(5): 411–416
selection was based on previous published articles with functional consideration (Nakada et al., 2011; Qinghai et al., 2014; Rasouli et al., 2013; Wang et al., 2012b; Yin et al., 2014; Zhang et al., 2013, 2014). Sample DNA (10 ng) were amplified by PCR according to the manufacturer’s recommendations. The SNP genotyping work was performed using a custom-by-design 48-Plex SNPscan Kit (Genesky Biotechnologies Inc., Shanghai, China) as previously described (Yin et al., 2014; Zheng et al., 2013). This kit was developed according to patented SNP genotyping technology by Genesky Biotechnologies Inc., which was based on double ligation and multiplex fluorescence PCR. For quality control, repeated analyses were done for 4% of randomly selected samples with high DNA quality. Furthermore, three samples with different genotypes for each SNP were selected to confirm the genotyping results by direct sequencing. Statistical analyses Differences in the distributions of demographic characteristics, selected variables, and genotypes of the IL17A rs2275913 G4A, rs3819024 A4G, rs3819025 G4A, rs4711998 A4G, rs8193036 C4T and rs8193037 G4A variants between the cases and controls were evaluated using the 2 test. The associations between IL17A rs2275913 G4A, rs3819024 A4G, rs3819025 G4A, rs4711998 A4G, rs8193036 C4T and rs8193037 G4A genotypes and risk of GCA were estimated by computing the ORs and their 95% CIs using logistic regression analyses for crude ORs and adjusted ORs when adjusting for age, sex, smoking and drinking status. The Hardy–Weinberg equilibrium (HWE) was tested by a goodness-of-fit 2 test to compare the observed genotype frequencies to the expected ones among the control subjects. All statistical analyses were performed with SAS 9.1.3 (SAS Institute, Cary, NC).
Results Characteristics of the study population Characteristics of cases and controls included in the study are summarized in Table 1. The cases and controls appeared to be adequately matched on age and sex as suggested by the 2 tests (p ¼ 0.923 and p ¼ 0.197, respectively). As shown in Table 1, no significant difference was detected on drinking status between the cases and the controls (p ¼ 0.217), but smoking rate was higher in GCA patients than in control subjects (p ¼ 0.004). Lymph node metastasis data was available in 220 (90.5%) of 243 GCA patients; regional lymph node metastasis was present in 122 (55.5%) cases. TNM stage data was available in 196 (80.7%) of 243 patients (stage I: 24; stage II: 34; stage III: 101, stage IV: 37) (Table 1). The primary information for six genotyped SNPs was in Table 2. For the six SNPs, the genotyping was successful ranging from 96.80% to 97.50% in all 719 samples. The concordance rates of repeated analyses were 100%. Minor allele frequency (MAF) in our controls was similar to MAF for Chinese in database for all six SNPs (Table 2). The observed genotype frequencies for these six polymorphisms in the controls were all in HWE except IL17A rs3819025 G4A (p ¼ 0.011) and IL17A rs4711998 A4G (p ¼ 0.042) (Table 2).
IL17A polymorphism and gastric cardia adenocarcinoma risk
DOI: 10.3109/1354750X.2014.924158
Associations between six polymorphisms and risk of GCA
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The genotype distributions of IL17A rs2275913 G4A, rs3819024 A4G, rs3819025 G4A, rs4711998 A4G, rs8193036 C4T and rs8193037 G4A in the cases and the controls are shown in Table 3. In the single locus analyses, the genotype frequencies of rs3819024 A4G were 25.53% (AA), 57.02% (AG), and 17.45% (GG) in the case patients and 31.55% (AA), 45.49% (AG), and 22.96% (GG) in the control subjects, and the difference was statistically significant (p ¼ 0.015). When the IL17A rs3819024 AA homozygote genotype was used as the reference group, the AG genotype was associated with a significantly increased risk for GCA (AG versus AA: adjusted OR ¼ 1.53, 95% CI ¼ 1.05–2.23, p ¼ 0.026). When the IL17A rs3819024 AA homozygote genotype was used as the reference group, the GG genotype Table 1. Distribution of selected demographic variables and risk factors in gastric cardia adenocarcinoma (GCA) cases and controls. Variable
Cases (n ¼ 243) N
Controls (n ¼ 476)
%
Age (years) 565 126 51.9 65 117 48.1 Age (years) Mean ± SD 64.90 (±8.65) Sex Male 159 65.4 Female 84 34.6 Tobacco use Never 144 59.3 Ever 99 40.7 Alcohol use Never 167 68.7 Ever 76 31.3 Lymph node metastasisb LN meta (+) 122 55.5 LN meta () 98 44.5 TNM stagesc I 24 12.2 II 34 17.3 III 101 51.5 IV 37 18.9
n
%
245 231
51.5 48.5
pa 0.923
64.76 (±7.46) 288 188
60.5 39.5
333 143
70.0 30.0
348 128
73.1 26.9
0.832 0.197 0.004 0.217
a
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was not associated with the risk for GCA (GG versus AA: adjusted OR ¼ 0.91, 95% CI ¼ 0.57–1.46, p ¼ 0.697). In the recessive model, when the IL17A rs3819024 AA/AG genotypes were used as the reference group, the GG homozygote genotype was not associated with the risk for GCA (adjusted OR ¼ 0.69, 95% CI ¼ 0.46–1.04, p ¼ 0.075). In the dominant model, the IL17A rs3819024 AG/GG variants were not associated with the risk of GCA, compared with the IL17A rs3819024 AA genotype (adjusted OR ¼ 1.32, 95% CI ¼ 0.92–1.88, p ¼ 0.128) (Table 3). None of the IL17A rs2275913 G4A, rs3819025 G4A, rs4711998 A4G, rs8193036 C4T and rs8193037 G4A polymorphisms achieved a significant difference in the genotype distributions between cases and controls (Table 3). Logistic regression analyses revealed that the five polymorphisms were not associated with the risk of GCA (Table 3). Stratification analyses of IL17A rs3819024 A4G polymorphisms and risk of GCA To evaluate the effects of IL17A rs3819024 A4G genotypes on GCA risk according to different age, sex, smoking and alcohol drinking status, we performed the stratification analyses (Table 4). A significantly increased risk of GCA associated with the IL17A rs3819024 A4G polymorphism was evident among male patients (AG versus AA: adjusted OR ¼ 1.95, 95% CI ¼ 1.19–3.17, p ¼ 0.008; AG/GG versus AA: adjusted OR ¼ 1.67, 95% CI ¼ 1.05–2.66, p ¼ 0.029), patients who drinking (AG versus AA: adjusted OR ¼ 2.24, 95% CI ¼ 1.03–4.87, p ¼ 0.042) or never smoking (AG versus AA: adjusted OR ¼ 1.61, 95% CI ¼ 1.01–2.58, p ¼ 0.045) (Table 4).
Discussion
Two-sided 2 test and student t test; Bold values are statistically significant (p50.05). b LN meta: Lymph Node Metastasis, LN information was available in 220 GCA cases. c TNM: tumor, nodal, and metastatic stage, TNM information was avaliable in 196 GCA cases.
In this hospital-based case–control study of GCA, we investigated the associations of IL17A rs2275913 G4A, rs3819024 A4G, rs3819025 G4A, rs4711998 A4G, rs8193036 C4T and rs8193037 G4A SNPs with GCA in a high-risk Chinese population. Our multivariate logistic analysis revealed that the IL17A rs3819024 AG genotype was associated with an increased risk of GCA. IL17A is the key cytokine of Th17 cells and induces proinflammatory cytokines. IL17A exhibits multiple biological activities on a variety of cells. Various tumor tissues overexpress IL17A, including multiple myeloma, ovarian
Table 2. Primary information for six genotyped SNPs.
Genotyped SNPs IL17A: IL17A: IL17A: IL17A: IL17A: IL17A: a
rs2275913 rs3819024 rs3819025 rs4711998 rs8193036 rs8193037
G4A A4G G4A A4G C4T G4A
Chr
Regulome DB Scorea
TFBSb
Location
MAFc for Chinese in database
MAF in our controls (n ¼ 476)
p Value for HWEd test in our controls
% Genotyping value
6 6 6 6 6 6
No data No data 5 No data No data No data
Y Y Y Y Y Y
50 -Flanking 50 -Flanking Intron1 50 -Flanking 50 -Flanking 50 -Flanking
0.465 0.477 0.159 0.244 0.265 0.082
0.444 0.457 0.174 0.273 0.285 0.120
0.256 0.072 0.011 0.042 0.084 0.578
97.50 97.50 97.36 96.94 96.80 97.50
http://www.regulomedb.org/. TFBS, Transcription Factor Binding Site (http://snpinfo.niehs.nih.gov/snpinfo/snpfunc.htm). c MAF, minor allele frequency. d HWE, Hardy–Weinberg equilibrium. b
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Biomarkers, 2014; 19(5): 411–416
Table 3. Logistic regression analyses of associations between IL17A polymorphisms and risk of GCA.
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Genotype IL17A: rs2275913 GG GA AA GA + AA GG + GA AA G allele A allele IL17A: rs3819024 AA AG GG AG + GG AA + AG GG A allele G allele IL17A: rs3819025 GG GA AA GA + AA GG + GA AA G allele A allele IL17A: rs4711998 AA AG GG AG + GG AA + AG GG A allele G allele IL17A: rs8193036 CC CT TT CT + TT CC + CT TT C allele T allele IL17A: rs8193037 GG GA AA GA + AA GG + GA AA G allele A allele
Cases (n ¼ 243)
Controls (n ¼ 476)
n
%
n
%
28.51 53.62 17.87 71.49 82.13 17.87 55.32 44.68
150 218 98 316 368 98 518 414
32.19 46.78 21.03 67.81 78.97 21.03 55.58 44.42
1.00 1.29 0.96 1.19 1.00 0.82 1.00 1.01
25.53 57.02 17.45 74.47 82.55 17.45 54.04 45.96
147 212 107 319 359 107 506 426
31.55 45.49 22.96 68.45 77.04 22.96 54.29 45.71
1.00 1.55 0.94 1.34 1.00 0.71 1.00 1.01
67.66 30.64 1.70 32.34 98.30 1.70 82.98 17.02
325 118 22 140 443 22 768 162
69.89 25.38 4.73 30.11 95.27 4.73 82.58 17.42
1.00 1.25 0.37 1.11 1.00 0.35 1.00 0.97
50.43 40.09 9.48 49.57 90.52 9.48 70.47 29.53
237 202 26 228 439 26 676 254
50.97 43.44 5.59 49.03 94.41 5.59 72.69 27.31
1.00 0.93 1.71 1.02 1.00 1.77 1.00 1.12
48.72 43.16 8.12 51.28 91.88 8.12 70.30 29.70
244 173 45 218 417 45 661 263
52.81 37.45 9.74 47.19 90.26 9.74 71.54 28.46
1.00 1.25 0.90 1.18 1.00 0.82 1.00 1.06
76.60 22.13 1.28 23.40 98.72 1.28 87.66 12.34
362 96 8 104 458 8 820 112
77.68 20.60 1.72 22.32 98.28 1.72 87.98 12.02
1.00 1.09 0.75 1.06 1.00 0.74 1.00 1.03
G4A 67 126 42 168 193 42 260 210 A4G 60 134 41 175 194 41 254 216 G4A 159 72 4 76 231 4 390 80 A4G 117 93 22 115 210 22 327 137 C4T 114 101 19 120 215 19 329 139 G4A 180 52 3 55 232 3 412 58
Crude OR (95%CI)
p
(0.90–1.86) (0.60–1.52) (0.84–1.68)
0.163 0.861 0.320
(0.55–1.22)
0.324
(0.81–1.26)
0.926
(1.07–2.24) (0.59–1.50) (0.95–1.91)
0.021 0.792 0.100
(0.48–1.06)
0.092
(0.81–1.26)
0.930
(0.88–1.77) (0.13–1.10) (0.79–1.56)
0.215 0.073 0.546
(0.12–1.02)
0.055
(0.73–1.31)
0.853
(0.67–1.30) (0.93–3.15) (0.75–1.40)
0.679 0.083 0.894
(0.98–3.20)
0.059
(0.87–1.43)
0.386
(0.90–1.74) (0.51–1.62) (0.86–1.61)
0.187 0.733 0.307
(0.47–1.44)
0.485
(0.83–1.36)
0.629
(0.74–1.60) (0.20–2.88) (0.73–1.54)
0.661 0.680 0.746
(0.20–2.82)
0.659
(0.74–1.45)
0.861
Adjusted ORa (95%CI) 1.00 1.30 0.93 1.18 1.00 0.79
1.00 1.53 0.91 1.32 1.00 0.69
1.00 1.28 0.40 1.14 1.00 0.37
1.00 0.95 1.70 1.03 1.00 1.74
1.00 1.24 0.94 1.18 1.00 0.86
1.00 1.11 0.89 1.09 1.00 0.87
p
(0.90–1.87) (0.59–1.49) (0.84–1.67)
0.165 0.770 0.346
(0.53–1.19)
0.262
(1.05–2.23) (0.57–1.46) (0.92–1.88)
0.026 0.697 0.128
(0.46–1.04)
0.075
(0.90–1.82) (0.13–1.17) (0.81–1.61)
0.173 0.094 0.448
(0.13–1.08)
0.070
(0.68–1.32) (0.92–3.15) (0.75–1.42)
0.750 0.091 0.835
(0.96–3.17)
0.069
(0.89–1.73) (0.53–1.69) (0.86–1.62)
0.210 0.844 0.310
(0.49–1.51)
0.595
(0.75–1.63) (0.23–3.41) (0.75–1.59)
0.608 0.862 0.650
(0.23–3.32)
0.836
a
Adjusted for age, sex, smoking and drinking status; Bold values are statistically significant (p50.05).
cancers, gastric cancer and breast cancer (Alexandrakis et al., 2006; Kato et al., 2001; Zhang et al., 2008; Zhu et al., 2008). IL17A polymorphisms were also associated with the risk of esophageal cancer, gastric cancer, breast cancer and cervical cancer (Quan et al., 2012; Shibata et al., 2009; Wang et al., 2012a; Yin et al., 2014). The IL17A rs3819024 A4G polymorphism might be associated with allergic rhinitis and comorbid asthma in Chinese population; however, no positive
association was found between the IL17A rs3819024 A4G SNP and cancers until now. We found that the IL17A rs3819024 AG heterozygote rather than the IL17A rs3819024 GG homozygote was associated with GCA. This might because our sample size was relatively small. Furthermore, the function of the IL17A rs3819024 A4G SNP remains to be investigated. Previous case–control reports of IL17A and gastric cancer risk mainly focused on IL17A
IL17A polymorphism and gastric cardia adenocarcinoma risk
The genotyping was successful in 235 (96.7%) GCA cases, and 466 (97.9%) controls for IL17A rs3819024 A4G. Adjusted for age, sex, smoking status and alcohol consumption (besides stratified factors accordingly) in a logistic regression model; Bold values are statistically significant (p50.05). b
a
0.63 (0.38–1.03); p: 0.066 0.76 (0.35–1.64); p: 0.478 1.19 (0.78–1.82); p: 0.411 1.98 (0.93–4.21); p: 0.076 0.78 (0.44–1.38); p: 0.387 1.36 (0.51–3.59); p: 0.536 1.43 (0.91–2.23); p: 0.119 2.24 (1.03–4.87); p: 0.042 1.00 1.00 28/80 13/27 87/148 47/64
115/228 60/91
0.63 (0.37–1.07); p: 0.088 0.78 (0.40–1.52); p: 0.459 0.85 (0.46–1.56); p: 0.595 1.02 (0.45–2.30); p: 0.958 22/75 19/32 79/141 55/71
101/216 74/103
1.00 1.00
1.61 (1.01–2.58); p: 0.045 1.51 (0.77–2.95); p: 0.229
1.35 (0.86–2.11); p: 0.188 1.34 (0.71–2.54); p: 0.363
0.67 (0.38–1.17); p: 0.159 0.73 (0.41–1.31); p: 0.291 1.25 (0.77–2.04); p: 0.368 1.45 (0.85–2.48); p: 0.174 0.85 (0.44–1.63); p: 0.615 1.02 (0.51–2.06); p: 0.947 1.47 (0.88–2.45); p: 0.144 1.68 (0.95–2.96); p: 0.072 1.00 1.00 21/55 20/52 70/104 64/108
91/159 84/160
0.75 (0.45–1.25); p: 0.275 0.50 (0.25–1.03); p: 0.059 1.67 (1.05–2.66); p: 0.029 0.84 (0.47–1.50); p: 0.554 1.16 (0.63–2.12); p: 0.635 0.51 (0.23–1.14); p: 0.101 1.95 (1.19–3.17); p: 0.008 1.02 (0.56–1.86); p: 0.958 28/62 13/45
Sex Male 34/95 Female 26/52 Age 565 34/79 65 26/68 Smoking status Never 38/110 Ever 22/37 Alcohol consumption Never 47/112 Ever 13/35
88/127 46/85
116/189 59/130
1.00 1.00
GG AG GG Variable
AA
AG
AG + GG
AA
Adjusted ORb (95% CI); p IL17A: rs3819024 A4G (case/control)a
Table 4. Stratified analyses between IL17A rs3819024 A4G polymorphism and GCA risk by sex, age, smoking status and alcohol consumption.
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AG + GG
GG versus (AG + AA)
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rs2275913 G4A (IL17A G-197A) SNP. Shibata et al. showed positive association between IL17A rs2275913 G4A SNP and gastric cancer risk (Arisawa et al., 2012; Shibata et al., 2009). Wu et al. found IL17A rs2275913 AG genotype was associated with increased risk of poorly differentiated, TNM I/II, age of 40–65-year subtypes of gastric cancer, but not with total gastric cancer risk (Wu et al., 2010). Zhang et al. found the rs2275913 G4A polymorphism increased gastric cancer risk, and interacted with Helicobacter pylori infection and subsites (Zhang et al., 2014). However, Rafiei et al. found the association was in early stage gastric adenocarcinomas only, and not linked to H. pylori infection (Rafiei et al., 2013). Kutikhin et al. found negative results, which was in accordance with our results (Kutikhin et al., 2014). It is possible that IL17A genes are associated with different degrees of genetic risk in different regions of gastric cancer and under different environmental conditions. Frequencies of genetic polymorphisms often vary between ethnic groups. In the present Chinese study, the allele frequency of IL17A rs3819024 G was 0.457 among 476 control subjects, which is consistent with that of the Chinese Han population (0.477) and Japanese (0.448) population in the SNP DataBase, but significantly higher than that of the Sub-Saharan African (0.296) population (http:// www.ncbi.nlm.nih.gov/SNP). Considering IL17A rs3819024 A4G mutant alleles in the control group, OR, GCA samples and control samples, the power of our analysis ( ¼ 0.05) was 0.767 in 243 GCA cases and 476 controls with an adjusted OR of 1.53 for IL17A rs3819024 A4G. Several limitations need to be addressed. First, the patients and controls may not represent the general population: inherited biases may be present. Second, the polymorphisms investigated in our study were chosen based on their functional considerations, and may not provide a comprehensive view of the genetic variability in IL17A. Further finemapping studies are needed. Third, the moderate sample size limited the statistical power of our study. Further studies in a large cohort are warranted to confirm our findings. Finally, detailed information on cancer metastasis and survival information was not available, which restricted further analyses of IL17A polymorphisms and GCA progression and prognosis. In conclusion, our study provides evidence that the functional polymorphism IL17A rs3819024 A4G may contribute to the risk of GCA. Our study was a preliminary investigation and future larger studies with more rigorous study designs of other ethnic populations are required to confirm the current findings.
Acknowledgements We wish to thank Dr. Da Ding and Dr. Yan Liu (Genesky Biotechnologies Inc., Shanghai, China) for technical support.
Declaration of interest This study was supported in part by National Natural Science Foundation of China (81370001, 81371927, 81101889, 81000028), Grants from Innovation Team of Jiangsu
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Province Hospital, Jiangsu Province Natural Science Foundation (BK2010333, BK2011481), Social Development Foundation of Zhenjiang (SH2010017) and Changzhou Young Talents and Science-Technology Foundation of Health Bureau (QN201102) and Affiliated People’s Hospital of Jiangsu University fund (Y200913). The authors declare that there is no conflict of interest.
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ISSN: 1354-750X (Print) 1366-5804 (Online) Journal homepage: http://www.tandfonline.com/loi/ibmk20
Interleukin 17A rs3819024 A>G polymorphism is associated with an increased risk of gastric cardia adenocarcinoma in a Chinese population Zhengbing Ren, Mingna Li, Ruiping Liu, Yong Wang & Haiyong Gu To cite this article: Zhengbing Ren, Mingna Li, Ruiping Liu, Yong Wang & Haiyong Gu (2014) Interleukin 17A rs3819024 A>G polymorphism is associated with an increased risk of gastric cardia adenocarcinoma in a Chinese population, Biomarkers, 19:5, 411-416 To link to this article: http://dx.doi.org/10.3109/1354750X.2014.924158
Published online: 04 Jun 2014.
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Date: 12 September 2015, At: 13:35
http://informahealthcare.com/bmk ISSN: 1354-750X (print), 1366-5804 (electronic) Biomarkers, 2014; 19(5): 411–416 ! 2014 Informa UK Ltd. DOI: 10.3109/1354750X.2014.924158
RESEARCH ARTICLE
Interleukin 17A rs3819024 A 4 G polymorphism is associated with an increased risk of gastric cardia adenocarcinoma in a Chinese population Zhengbing Ren1*, Mingna Li2*, Ruiping Liu3, Yong Wang4#, and Haiyong Gu1# Department of Cardiothoracic Surgery, Affiliated People’s Hospital of Jiangsu University, Zhenjiang, China, 2Department of Pathology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China, 3Department of Orthopedics, Affiliated Hospital of Nanjing Medical University, Changzhou Second People’s Hospital, Changzhou, China, and 4Department of Cardiothoracic Surgery, Affiliated Hospital of Nanjing Medical University, Changzhou Second People’s Hospital, Changzhou, China
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1
Abstract
Keywords
Gastric cardia adenocarcinoma (GCA) is one of the most common malignant tumors. In addition to environmental risk factors, genetic factors might play an important role in GCA carcinogenesis. To evaluate the association between polymorphisms in the interleukin 17A (IL17A) gene on the development of GCA, we conducted a hospital-based case–control study. A total of 243 GCA cases and 476 controls were recruited and their genotypes were determined using a custom-by-design 48-Plex SNPscanä Kit. IL17A rs3819024 A4G polymorphism was found to be associated with the increased risk of GCA. When the IL17A rs3819024 AA homozygote genotype was used as the reference group, the AG genotype was associated with a significantly increased risk of GCA (AG versus AA: adjusted OR ¼ 1.53, 95% CI ¼ 1.05–2.23, p ¼ 0.026). However, there was no significant association between five other SNPs and GCA. Stratified analyses indicated that a significantly increased risk of GCA associated with the IL17A rs3819024 A4G polymorphism was evident among male patients, patients who drank alcohol or those who never smoked. These findings indicated that functional polymorphism IL17A rs3819024 A4G might contribute to GCA susceptibility. Future larger studies with more rigorous study designs are required to confirm the current findings.
Gastric cardia adenocarcinoma, IL17A, molecular epidemiology, polymorphisms
Introduction Gastric cardia adenocarcinoma (GCA) is one of the most common malignant tumors worldwide (Zhang et al., 2010). GCA has a multi-factorial etiology: both genetic and environmental factors are suspected to contribute to GCA development. Besides smoking and heavy drinking, single nucleotide polymorphisms (SNPs) might partly explain individual differences in GCA susceptibility (Zhang et al., 2012). Infection and chronic inflammation contribute to about 1/4 of all cancer cases (Hussain & Harris, 2007). Cytokines are key regulators to promote or restrain inflammation, and therefore, can either stimulate or inhibit tumor growth and progression (Lin & Karin, 2007; Smyth et al., 2004).
*Zhengbing Ren and Mingna Li contributed equally to this work. # Yong Wang and Haiyong Gu are responsible for statistical design/ analysis. E-mail: [email protected]; haiyong_gu@hotmail. com. Address for correspondence: Professor Yong Wang, Department of Cardiothoracic Surgery, Changzhou Second People’s Hospital, Affiliated Hospital of Nanjing Medical University, Changzhou 213003, China. E-mail: [email protected] Dr. Haiyong Gu, Department of Cardiothoracic Surgery, Affiliated People’s Hospital of Jiangsu University, Zhenjiang 212000, China. E-mail: [email protected]
History Received 31 March 2014 Revised 9 May 2014 Accepted 11 May 2014 Published online 4 June 2014
Interleukin 17A (IL17A), also known as IL17, is the signature T helper 17 (Th17) cell effector cytokine (Korn et al., 2009). IL17A plays an important role in inflammation and leads to the induction of many proinflammatory factors, including tumor necrosis factor-alpha (TNF-a), IL6 and IL1b (Ruddy et al., 2004). High expression of IL17A was observed in various tumor tissues, including multiple myeloma, gastric cancer and breast cancer (Alexandrakis et al., 2006; Zhang et al., 2008; Zhu et al., 2008). Through stimulation of vascular endothelial cell migration and regulation of a series of proangiogenic factors, IL17A enhances angiogenesis (Numasaki et al., 2003). IL17A also mediates tissue remodeling by stimulating the production of angiogenic factors and matrix metalloproteases (Jovanovic et al., 2000). IL17A contributes to the development and progression of prostate cancer, gastric carcinomas and skin cancer (Chen et al., 2011; He et al., 2012; Steiner et al., 2003). IL17A polymorphisms have been associated with the risk of various types of cancer, including esophageal cancer, gastric cancer, breast cancer and cervical cancer (Arisawa et al., 2012; Quan et al., 2012; Wang et al., 2012a; Yin et al., 2014). The IL17A gene is located on human chromosome 6p12.1. The wellstudied IL17A rs2275913 G4A polymorphism is located within a binding motif for nuclear factor activated T cells and
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plays a functional role in the promoter activity of the IL17A gene (Liu et al., 2004). We previously conducted a hospital-based case–control study to evaluate IL17A rs2275913 G4A, rs3819024 A4G, rs3819025 G4A, rs4711998 A4G, rs8193036 C4T and rs8193037 G4A polymorphisms and esophageal squamous cell carcinoma (ESCC) risk (Yin et al., 2014). We found that the IL17A rs4711998 A4G AG genotype was associated with a significantly decreased risk of ESCC (Yin et al., 2014). In this investigation, we evaluated the association between the six functional SNPs and GCA susceptibility in 243 GCA cases and 476 controls.
Patients and methods
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Ethical approval of the study protocol This hospital-based case–control study was approved by the Review Board of Jiangsu University (Zhenjiang, China). We have complied with the World Medical Association Declaration of Helsinki regarding ethical conduct of research involving human subjects and/or animals. All subjects provided written informed consent to be included in the study. Study subjects Two-hundred and forty-three subjects with gastric cardia cancer were consecutively recruited from the Affiliated People’s Hospital of Jiangsu University and Affiliated Hospital of Jiangsu University (Zhenjiang, China) between October 2008 and July 2010. All cases of gastric cardia cancer were diagnosed as GCA by pathologic means. The exclusion criteria were patients who previously had: cancer; any metastasized cancer; radiotherapy or chemotherapy. Among 476 controls, 380 controls were patients without cancer frequency-matched to the cases with regard to age (±5 years) and sex recruited from the two hospitals mentioned above during the same time period. Another 96 controls were recruited from hospitals in Changzhou city (which is a neighboring city of Zhenjiang) as previous described (Cheng et al., 2012). Most of the controls were admitted to the hospitals for the treatment of trauma. Each subject was personally questioned by trained interviewers using a pre-tested questionnaire to obtain information on demographic data (e.g. age, sex) and related risk factors (including tobacco smoking and alcohol consumption). After the interview, 2-mL samples of venous blood were collected from each subject. Individuals who smoked one cigarette per day for 41 year were defined as ‘‘smokers’’. Subjects who consumed 3 alcoholic drinks a week for 46 months were considered to be ‘‘alcohol drinkers’’. Tumor, nodal, and metastatic (TNM) stage was according to the seventh edition of the UICC TNM staging (Kwon, 2011). Isolation of DNA, SNPs selection and genotyping by a custom-by-design 48-Plex SNPscan Kit Blood samples were collected from patients using Vacutainers and transferred to tubes lined with ethylenediamine tetraacetic acid (EDTA). Genomic DNA was isolated from whole blood with the QIAamp DNA Blood Mini Kit (Qiagen, Hilden, Germany) (Zheng et al., 2013). The IL17A SNPs
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selection was based on previous published articles with functional consideration (Nakada et al., 2011; Qinghai et al., 2014; Rasouli et al., 2013; Wang et al., 2012b; Yin et al., 2014; Zhang et al., 2013, 2014). Sample DNA (10 ng) were amplified by PCR according to the manufacturer’s recommendations. The SNP genotyping work was performed using a custom-by-design 48-Plex SNPscan Kit (Genesky Biotechnologies Inc., Shanghai, China) as previously described (Yin et al., 2014; Zheng et al., 2013). This kit was developed according to patented SNP genotyping technology by Genesky Biotechnologies Inc., which was based on double ligation and multiplex fluorescence PCR. For quality control, repeated analyses were done for 4% of randomly selected samples with high DNA quality. Furthermore, three samples with different genotypes for each SNP were selected to confirm the genotyping results by direct sequencing. Statistical analyses Differences in the distributions of demographic characteristics, selected variables, and genotypes of the IL17A rs2275913 G4A, rs3819024 A4G, rs3819025 G4A, rs4711998 A4G, rs8193036 C4T and rs8193037 G4A variants between the cases and controls were evaluated using the 2 test. The associations between IL17A rs2275913 G4A, rs3819024 A4G, rs3819025 G4A, rs4711998 A4G, rs8193036 C4T and rs8193037 G4A genotypes and risk of GCA were estimated by computing the ORs and their 95% CIs using logistic regression analyses for crude ORs and adjusted ORs when adjusting for age, sex, smoking and drinking status. The Hardy–Weinberg equilibrium (HWE) was tested by a goodness-of-fit 2 test to compare the observed genotype frequencies to the expected ones among the control subjects. All statistical analyses were performed with SAS 9.1.3 (SAS Institute, Cary, NC).
Results Characteristics of the study population Characteristics of cases and controls included in the study are summarized in Table 1. The cases and controls appeared to be adequately matched on age and sex as suggested by the 2 tests (p ¼ 0.923 and p ¼ 0.197, respectively). As shown in Table 1, no significant difference was detected on drinking status between the cases and the controls (p ¼ 0.217), but smoking rate was higher in GCA patients than in control subjects (p ¼ 0.004). Lymph node metastasis data was available in 220 (90.5%) of 243 GCA patients; regional lymph node metastasis was present in 122 (55.5%) cases. TNM stage data was available in 196 (80.7%) of 243 patients (stage I: 24; stage II: 34; stage III: 101, stage IV: 37) (Table 1). The primary information for six genotyped SNPs was in Table 2. For the six SNPs, the genotyping was successful ranging from 96.80% to 97.50% in all 719 samples. The concordance rates of repeated analyses were 100%. Minor allele frequency (MAF) in our controls was similar to MAF for Chinese in database for all six SNPs (Table 2). The observed genotype frequencies for these six polymorphisms in the controls were all in HWE except IL17A rs3819025 G4A (p ¼ 0.011) and IL17A rs4711998 A4G (p ¼ 0.042) (Table 2).
IL17A polymorphism and gastric cardia adenocarcinoma risk
DOI: 10.3109/1354750X.2014.924158
Associations between six polymorphisms and risk of GCA
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The genotype distributions of IL17A rs2275913 G4A, rs3819024 A4G, rs3819025 G4A, rs4711998 A4G, rs8193036 C4T and rs8193037 G4A in the cases and the controls are shown in Table 3. In the single locus analyses, the genotype frequencies of rs3819024 A4G were 25.53% (AA), 57.02% (AG), and 17.45% (GG) in the case patients and 31.55% (AA), 45.49% (AG), and 22.96% (GG) in the control subjects, and the difference was statistically significant (p ¼ 0.015). When the IL17A rs3819024 AA homozygote genotype was used as the reference group, the AG genotype was associated with a significantly increased risk for GCA (AG versus AA: adjusted OR ¼ 1.53, 95% CI ¼ 1.05–2.23, p ¼ 0.026). When the IL17A rs3819024 AA homozygote genotype was used as the reference group, the GG genotype Table 1. Distribution of selected demographic variables and risk factors in gastric cardia adenocarcinoma (GCA) cases and controls. Variable
Cases (n ¼ 243) N
Controls (n ¼ 476)
%
Age (years) 565 126 51.9 65 117 48.1 Age (years) Mean ± SD 64.90 (±8.65) Sex Male 159 65.4 Female 84 34.6 Tobacco use Never 144 59.3 Ever 99 40.7 Alcohol use Never 167 68.7 Ever 76 31.3 Lymph node metastasisb LN meta (+) 122 55.5 LN meta () 98 44.5 TNM stagesc I 24 12.2 II 34 17.3 III 101 51.5 IV 37 18.9
n
%
245 231
51.5 48.5
pa 0.923
64.76 (±7.46) 288 188
60.5 39.5
333 143
70.0 30.0
348 128
73.1 26.9
0.832 0.197 0.004 0.217
a
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was not associated with the risk for GCA (GG versus AA: adjusted OR ¼ 0.91, 95% CI ¼ 0.57–1.46, p ¼ 0.697). In the recessive model, when the IL17A rs3819024 AA/AG genotypes were used as the reference group, the GG homozygote genotype was not associated with the risk for GCA (adjusted OR ¼ 0.69, 95% CI ¼ 0.46–1.04, p ¼ 0.075). In the dominant model, the IL17A rs3819024 AG/GG variants were not associated with the risk of GCA, compared with the IL17A rs3819024 AA genotype (adjusted OR ¼ 1.32, 95% CI ¼ 0.92–1.88, p ¼ 0.128) (Table 3). None of the IL17A rs2275913 G4A, rs3819025 G4A, rs4711998 A4G, rs8193036 C4T and rs8193037 G4A polymorphisms achieved a significant difference in the genotype distributions between cases and controls (Table 3). Logistic regression analyses revealed that the five polymorphisms were not associated with the risk of GCA (Table 3). Stratification analyses of IL17A rs3819024 A4G polymorphisms and risk of GCA To evaluate the effects of IL17A rs3819024 A4G genotypes on GCA risk according to different age, sex, smoking and alcohol drinking status, we performed the stratification analyses (Table 4). A significantly increased risk of GCA associated with the IL17A rs3819024 A4G polymorphism was evident among male patients (AG versus AA: adjusted OR ¼ 1.95, 95% CI ¼ 1.19–3.17, p ¼ 0.008; AG/GG versus AA: adjusted OR ¼ 1.67, 95% CI ¼ 1.05–2.66, p ¼ 0.029), patients who drinking (AG versus AA: adjusted OR ¼ 2.24, 95% CI ¼ 1.03–4.87, p ¼ 0.042) or never smoking (AG versus AA: adjusted OR ¼ 1.61, 95% CI ¼ 1.01–2.58, p ¼ 0.045) (Table 4).
Discussion
Two-sided 2 test and student t test; Bold values are statistically significant (p50.05). b LN meta: Lymph Node Metastasis, LN information was available in 220 GCA cases. c TNM: tumor, nodal, and metastatic stage, TNM information was avaliable in 196 GCA cases.
In this hospital-based case–control study of GCA, we investigated the associations of IL17A rs2275913 G4A, rs3819024 A4G, rs3819025 G4A, rs4711998 A4G, rs8193036 C4T and rs8193037 G4A SNPs with GCA in a high-risk Chinese population. Our multivariate logistic analysis revealed that the IL17A rs3819024 AG genotype was associated with an increased risk of GCA. IL17A is the key cytokine of Th17 cells and induces proinflammatory cytokines. IL17A exhibits multiple biological activities on a variety of cells. Various tumor tissues overexpress IL17A, including multiple myeloma, ovarian
Table 2. Primary information for six genotyped SNPs.
Genotyped SNPs IL17A: IL17A: IL17A: IL17A: IL17A: IL17A: a
rs2275913 rs3819024 rs3819025 rs4711998 rs8193036 rs8193037
G4A A4G G4A A4G C4T G4A
Chr
Regulome DB Scorea
TFBSb
Location
MAFc for Chinese in database
MAF in our controls (n ¼ 476)
p Value for HWEd test in our controls
% Genotyping value
6 6 6 6 6 6
No data No data 5 No data No data No data
Y Y Y Y Y Y
50 -Flanking 50 -Flanking Intron1 50 -Flanking 50 -Flanking 50 -Flanking
0.465 0.477 0.159 0.244 0.265 0.082
0.444 0.457 0.174 0.273 0.285 0.120
0.256 0.072 0.011 0.042 0.084 0.578
97.50 97.50 97.36 96.94 96.80 97.50
http://www.regulomedb.org/. TFBS, Transcription Factor Binding Site (http://snpinfo.niehs.nih.gov/snpinfo/snpfunc.htm). c MAF, minor allele frequency. d HWE, Hardy–Weinberg equilibrium. b
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Table 3. Logistic regression analyses of associations between IL17A polymorphisms and risk of GCA.
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Genotype IL17A: rs2275913 GG GA AA GA + AA GG + GA AA G allele A allele IL17A: rs3819024 AA AG GG AG + GG AA + AG GG A allele G allele IL17A: rs3819025 GG GA AA GA + AA GG + GA AA G allele A allele IL17A: rs4711998 AA AG GG AG + GG AA + AG GG A allele G allele IL17A: rs8193036 CC CT TT CT + TT CC + CT TT C allele T allele IL17A: rs8193037 GG GA AA GA + AA GG + GA AA G allele A allele
Cases (n ¼ 243)
Controls (n ¼ 476)
n
%
n
%
28.51 53.62 17.87 71.49 82.13 17.87 55.32 44.68
150 218 98 316 368 98 518 414
32.19 46.78 21.03 67.81 78.97 21.03 55.58 44.42
1.00 1.29 0.96 1.19 1.00 0.82 1.00 1.01
25.53 57.02 17.45 74.47 82.55 17.45 54.04 45.96
147 212 107 319 359 107 506 426
31.55 45.49 22.96 68.45 77.04 22.96 54.29 45.71
1.00 1.55 0.94 1.34 1.00 0.71 1.00 1.01
67.66 30.64 1.70 32.34 98.30 1.70 82.98 17.02
325 118 22 140 443 22 768 162
69.89 25.38 4.73 30.11 95.27 4.73 82.58 17.42
1.00 1.25 0.37 1.11 1.00 0.35 1.00 0.97
50.43 40.09 9.48 49.57 90.52 9.48 70.47 29.53
237 202 26 228 439 26 676 254
50.97 43.44 5.59 49.03 94.41 5.59 72.69 27.31
1.00 0.93 1.71 1.02 1.00 1.77 1.00 1.12
48.72 43.16 8.12 51.28 91.88 8.12 70.30 29.70
244 173 45 218 417 45 661 263
52.81 37.45 9.74 47.19 90.26 9.74 71.54 28.46
1.00 1.25 0.90 1.18 1.00 0.82 1.00 1.06
76.60 22.13 1.28 23.40 98.72 1.28 87.66 12.34
362 96 8 104 458 8 820 112
77.68 20.60 1.72 22.32 98.28 1.72 87.98 12.02
1.00 1.09 0.75 1.06 1.00 0.74 1.00 1.03
G4A 67 126 42 168 193 42 260 210 A4G 60 134 41 175 194 41 254 216 G4A 159 72 4 76 231 4 390 80 A4G 117 93 22 115 210 22 327 137 C4T 114 101 19 120 215 19 329 139 G4A 180 52 3 55 232 3 412 58
Crude OR (95%CI)
p
(0.90–1.86) (0.60–1.52) (0.84–1.68)
0.163 0.861 0.320
(0.55–1.22)
0.324
(0.81–1.26)
0.926
(1.07–2.24) (0.59–1.50) (0.95–1.91)
0.021 0.792 0.100
(0.48–1.06)
0.092
(0.81–1.26)
0.930
(0.88–1.77) (0.13–1.10) (0.79–1.56)
0.215 0.073 0.546
(0.12–1.02)
0.055
(0.73–1.31)
0.853
(0.67–1.30) (0.93–3.15) (0.75–1.40)
0.679 0.083 0.894
(0.98–3.20)
0.059
(0.87–1.43)
0.386
(0.90–1.74) (0.51–1.62) (0.86–1.61)
0.187 0.733 0.307
(0.47–1.44)
0.485
(0.83–1.36)
0.629
(0.74–1.60) (0.20–2.88) (0.73–1.54)
0.661 0.680 0.746
(0.20–2.82)
0.659
(0.74–1.45)
0.861
Adjusted ORa (95%CI) 1.00 1.30 0.93 1.18 1.00 0.79
1.00 1.53 0.91 1.32 1.00 0.69
1.00 1.28 0.40 1.14 1.00 0.37
1.00 0.95 1.70 1.03 1.00 1.74
1.00 1.24 0.94 1.18 1.00 0.86
1.00 1.11 0.89 1.09 1.00 0.87
p
(0.90–1.87) (0.59–1.49) (0.84–1.67)
0.165 0.770 0.346
(0.53–1.19)
0.262
(1.05–2.23) (0.57–1.46) (0.92–1.88)
0.026 0.697 0.128
(0.46–1.04)
0.075
(0.90–1.82) (0.13–1.17) (0.81–1.61)
0.173 0.094 0.448
(0.13–1.08)
0.070
(0.68–1.32) (0.92–3.15) (0.75–1.42)
0.750 0.091 0.835
(0.96–3.17)
0.069
(0.89–1.73) (0.53–1.69) (0.86–1.62)
0.210 0.844 0.310
(0.49–1.51)
0.595
(0.75–1.63) (0.23–3.41) (0.75–1.59)
0.608 0.862 0.650
(0.23–3.32)
0.836
a
Adjusted for age, sex, smoking and drinking status; Bold values are statistically significant (p50.05).
cancers, gastric cancer and breast cancer (Alexandrakis et al., 2006; Kato et al., 2001; Zhang et al., 2008; Zhu et al., 2008). IL17A polymorphisms were also associated with the risk of esophageal cancer, gastric cancer, breast cancer and cervical cancer (Quan et al., 2012; Shibata et al., 2009; Wang et al., 2012a; Yin et al., 2014). The IL17A rs3819024 A4G polymorphism might be associated with allergic rhinitis and comorbid asthma in Chinese population; however, no positive
association was found between the IL17A rs3819024 A4G SNP and cancers until now. We found that the IL17A rs3819024 AG heterozygote rather than the IL17A rs3819024 GG homozygote was associated with GCA. This might because our sample size was relatively small. Furthermore, the function of the IL17A rs3819024 A4G SNP remains to be investigated. Previous case–control reports of IL17A and gastric cancer risk mainly focused on IL17A
IL17A polymorphism and gastric cardia adenocarcinoma risk
The genotyping was successful in 235 (96.7%) GCA cases, and 466 (97.9%) controls for IL17A rs3819024 A4G. Adjusted for age, sex, smoking status and alcohol consumption (besides stratified factors accordingly) in a logistic regression model; Bold values are statistically significant (p50.05). b
a
0.63 (0.38–1.03); p: 0.066 0.76 (0.35–1.64); p: 0.478 1.19 (0.78–1.82); p: 0.411 1.98 (0.93–4.21); p: 0.076 0.78 (0.44–1.38); p: 0.387 1.36 (0.51–3.59); p: 0.536 1.43 (0.91–2.23); p: 0.119 2.24 (1.03–4.87); p: 0.042 1.00 1.00 28/80 13/27 87/148 47/64
115/228 60/91
0.63 (0.37–1.07); p: 0.088 0.78 (0.40–1.52); p: 0.459 0.85 (0.46–1.56); p: 0.595 1.02 (0.45–2.30); p: 0.958 22/75 19/32 79/141 55/71
101/216 74/103
1.00 1.00
1.61 (1.01–2.58); p: 0.045 1.51 (0.77–2.95); p: 0.229
1.35 (0.86–2.11); p: 0.188 1.34 (0.71–2.54); p: 0.363
0.67 (0.38–1.17); p: 0.159 0.73 (0.41–1.31); p: 0.291 1.25 (0.77–2.04); p: 0.368 1.45 (0.85–2.48); p: 0.174 0.85 (0.44–1.63); p: 0.615 1.02 (0.51–2.06); p: 0.947 1.47 (0.88–2.45); p: 0.144 1.68 (0.95–2.96); p: 0.072 1.00 1.00 21/55 20/52 70/104 64/108
91/159 84/160
0.75 (0.45–1.25); p: 0.275 0.50 (0.25–1.03); p: 0.059 1.67 (1.05–2.66); p: 0.029 0.84 (0.47–1.50); p: 0.554 1.16 (0.63–2.12); p: 0.635 0.51 (0.23–1.14); p: 0.101 1.95 (1.19–3.17); p: 0.008 1.02 (0.56–1.86); p: 0.958 28/62 13/45
Sex Male 34/95 Female 26/52 Age 565 34/79 65 26/68 Smoking status Never 38/110 Ever 22/37 Alcohol consumption Never 47/112 Ever 13/35
88/127 46/85
116/189 59/130
1.00 1.00
GG AG GG Variable
AA
AG
AG + GG
AA
Adjusted ORb (95% CI); p IL17A: rs3819024 A4G (case/control)a
Table 4. Stratified analyses between IL17A rs3819024 A4G polymorphism and GCA risk by sex, age, smoking status and alcohol consumption.
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AG + GG
GG versus (AG + AA)
DOI: 10.3109/1354750X.2014.924158
415
rs2275913 G4A (IL17A G-197A) SNP. Shibata et al. showed positive association between IL17A rs2275913 G4A SNP and gastric cancer risk (Arisawa et al., 2012; Shibata et al., 2009). Wu et al. found IL17A rs2275913 AG genotype was associated with increased risk of poorly differentiated, TNM I/II, age of 40–65-year subtypes of gastric cancer, but not with total gastric cancer risk (Wu et al., 2010). Zhang et al. found the rs2275913 G4A polymorphism increased gastric cancer risk, and interacted with Helicobacter pylori infection and subsites (Zhang et al., 2014). However, Rafiei et al. found the association was in early stage gastric adenocarcinomas only, and not linked to H. pylori infection (Rafiei et al., 2013). Kutikhin et al. found negative results, which was in accordance with our results (Kutikhin et al., 2014). It is possible that IL17A genes are associated with different degrees of genetic risk in different regions of gastric cancer and under different environmental conditions. Frequencies of genetic polymorphisms often vary between ethnic groups. In the present Chinese study, the allele frequency of IL17A rs3819024 G was 0.457 among 476 control subjects, which is consistent with that of the Chinese Han population (0.477) and Japanese (0.448) population in the SNP DataBase, but significantly higher than that of the Sub-Saharan African (0.296) population (http:// www.ncbi.nlm.nih.gov/SNP). Considering IL17A rs3819024 A4G mutant alleles in the control group, OR, GCA samples and control samples, the power of our analysis ( ¼ 0.05) was 0.767 in 243 GCA cases and 476 controls with an adjusted OR of 1.53 for IL17A rs3819024 A4G. Several limitations need to be addressed. First, the patients and controls may not represent the general population: inherited biases may be present. Second, the polymorphisms investigated in our study were chosen based on their functional considerations, and may not provide a comprehensive view of the genetic variability in IL17A. Further finemapping studies are needed. Third, the moderate sample size limited the statistical power of our study. Further studies in a large cohort are warranted to confirm our findings. Finally, detailed information on cancer metastasis and survival information was not available, which restricted further analyses of IL17A polymorphisms and GCA progression and prognosis. In conclusion, our study provides evidence that the functional polymorphism IL17A rs3819024 A4G may contribute to the risk of GCA. Our study was a preliminary investigation and future larger studies with more rigorous study designs of other ethnic populations are required to confirm the current findings.
Acknowledgements We wish to thank Dr. Da Ding and Dr. Yan Liu (Genesky Biotechnologies Inc., Shanghai, China) for technical support.
Declaration of interest This study was supported in part by National Natural Science Foundation of China (81370001, 81371927, 81101889, 81000028), Grants from Innovation Team of Jiangsu
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Z. Ren et al.
Province Hospital, Jiangsu Province Natural Science Foundation (BK2010333, BK2011481), Social Development Foundation of Zhenjiang (SH2010017) and Changzhou Young Talents and Science-Technology Foundation of Health Bureau (QN201102) and Affiliated People’s Hospital of Jiangsu University fund (Y200913). The authors declare that there is no conflict of interest.
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