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Effects of ALOX5, IL6R and SFTPD gene polymorphisms on the risk of lung cancer: A case-control study in China
International Immunopharmacology 79 (2020) 106155
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Effects of ALOX5, IL6R and SFTPD gene polymorphisms on the risk of lung cancer: A case-control study in China
T
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Xiaoping Wei , Chen Wang, Haiming Feng, Bing Li, Peng Jiang, Jianbao Yang, Duojie Zhu, Shaobo Zhang, Tao Jin, Yuqi Meng Department of Thoracic Surgery, Lanzhou University Second Hospital, Lanzhou, Gansu 730030, China
A R T I C LE I N FO
A B S T R A C T
Keywords: Lung cancer Polymorphisms Arachidonate 5-lipoxygenase (ALOX5) Interleukin 6 receptor (IL6R) Surfactant protein D (SFTPD)
Background: ALOX5, IL6R and SFTPD are all immune related genes that may be involved in the development of lung cancer. We sought to explore the effect of polymorphisms of these genes on the risk of lung cancer. Methods: Six single nucleotide polymorphisms (SNPs) were genotyped using a MassARRAY platform in a casecontrol cohort including 550 patients with lung cancer and 550 healthy controls. Results: The rs4845626-T and rs4329505-C alleles were associated with a decreased risk of lung cancer (p < 0.001), while the rs745986-G and rs2245121-A alleles were correlated with an increased risk of lung cancer (p < 0.01). The rs4845626-GT/GG and rs4329505-TC genotypes were protective against lung cancer (p < 0.001). However, the rs745986-AG and rs2245121-AG/AA genotypes were associated with an increased risk of lung cancer (p < 0.01). Stratification analysis showed that the rs4845626 and rs4329505 polymorphisms of IL6R were associated with a reduced risk of lung cancer in both smokers and nonsmokers (p < 0.05). However, rs892690, rs745986 and rs2115819 of ALOX5 were associated with an increased risk of disease in nonsmokers, while rs2245121 of SFTPD was correlated with a higher risk of disease in smokers (p < 0.05). Conclusion: Our results provide candidate SNPs for early screening for lung cancer and new clues for further study of the pathogenesis of the disease.
1. Introduction Lung cancer is a challenge for clinicians and researchers because of its high incidence and mortality, especially in developing countries, such as China [1]. Based on clinical experience, early detection and prevention of progression are crucial to prolong the survival time of patients with lung cancer [2]. In recent years, with the development of medical molecular biology, single nucleotide polymorphisms (SNPs) have been gradually found to have enormous potential in early screening and personalized treatment of lung cancer [3]. To date, a number of susceptibility genes and SNPs have been reported to have positive or negative correlations with the risk of developing lung cancer [4–6]. However, the current level of knowledge is insufficient for totally understanding the genetic background of lung cancer. Chronic inflammation is strongly associated with the proliferation, invasion and metastasis of tumor cells in the human body [7]. Moreover, polymorphisms in inflammation-related genes have been shown to be involved in the development of lung cancer [8,9]. These results drew our attention to the connection between polymorphisms in lung
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inflammation-related genes and the risk of lung cancer. Arachidonate 5lipoxygenase (ALOX5) is part of the leukotriene pathway and, therefore, plays important roles in the development of inflammation and inflammatory diseases [10]. ALOX5 polymorphisms have been associated with elevated levels of leukotriene, resulting in impairment of lung function [11]. However, few studies have directly investigated the correlation between ALOX5 polymorphisms and lung cancer risk. Interleukin 6 (IL6) polymorphisms have been intensively investigated in patients with lung cancer in several different populations [12–14], but the correlation between IL6 receptor (IL6R) polymorphisms and risk of disease is still unclear. Surfactant protein D (SFTPD) regulates inflammatory processes and protects the host against pathogens invading pulmonary alveolar type II cells, indicating it has a potential role in blocking the progression of lung cancer [15]. Therefore, the present study selected ALOX5, IL6R and SFTPD as candidate genes to investigate. rs4845626 and rs4329505 in IL6R have been associated with a decreased risk of COPD in Mexicans [16]. rs892690, rs745986 and
Corresponding author at: #82 Cuiyingmen, Lanzhou, Gansu 730030, China. E-mail addresses: [email protected], [email protected] (X. Wei).
https://doi.org/10.1016/j.intimp.2019.106155 Received 16 October 2019; Received in revised form 24 December 2019; Accepted 25 December 2019 1567-5769/ © 2020 Elsevier B.V. All rights reserved.
International Immunopharmacology 79 (2020) 106155
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rs2115819 in ALOX5 were shown to be associated with the metabolism of montelukast in patients with asthma [17]. Moreover, rs2245121 in SFTPD was found to be correlated with an increased risk of COPD [18]. In the present study, we sought to explore the association between these SNPs and the risk of lung cancer.
Table 1 The demographic characteristics of the participants. Variables
Sex (%) Male Female Age (mean ± SD), years BMI (mean ± SD), kg/ m2 Smoking (%) Ever Never Drinking (%) Ever Never Histology (%) Adenocarcinoma Squamous Other
2. Materials and methods 2.1. Participants All subjects are ethnic Chinese Han living in Lanzhou city or the surrounding areas. A total of 550 lung cancer patients and 550 healthy individuals were recruited for the study as a case and a control group, respectively. The cases were diagnosed with lung cancer by pathological biopsy at our hospital. Cases with any other types of cancers, immune diseases or mental disorders were excluded. The controls were recruited at the physical examination center of our hospital and included only those with no history of cancer, lung disease, or immune disease. We obtained written informed consent from each subject, and the study was approved by the Ethics Department of Lanzhou University Second Hospital.
Case (%) (n = 550)
Control (%) (n = 550)
302 (54.9) 248 (45.1) 58.2 ± 9.7
300 (54.5) 250 (45.5) 58.5 ± 10.2
24.3 ± 3.5
24.5 ± 3.8
255 (46.4) 295 (53.6)
209 (38.0) 341 (62.0)
224 (40.7) 326 (59.3)
232 (42.2) 318 (57.8)
χ 2/t
p
0.015
0.903
0.838
0.186
0.564
0.242
7.887
0.005*
0.240
0.624
232 (42.2) 224 (40.7) 94 (17.1)
(OR = 0.64, 95% CI = 0.54–0.76, p < 0.001). The allele C of rs4329505 also exhibited a protective effect against the risk of lung cancer (OR = 0.75, 95% CI = 0.63–0.90, p = 0.001). In contrast, the allele G of rs745986 was correlated with a 1.40-fold increased risk of lung cancer (95% CI = 1.09–1.79, p = 0.008), and the allele A of rs2245121 was also associated with an increased risk of disease (OR = 1.33, 95% CI = 1.13–1.58, p = 0.001). For the genotypes, both the GT and GG genotypes of rs4845626 were correlated with a decreased risk of lung cancer (p < 0.001). The TC genotype of rs4329505 also showed a protective effect against the risk of lung cancer (p < 0.001). However, the AG genotype of rs745986 (p = 0.006) and the AG and AA genotypes of rs2245121 (p = 0.003) were all associated with an increased risk of lung cancer. The associations between genotypes and the risk of lung cancer were also evaluated based on genetic models (Table 4). All of the SNPs were identified as having associations with the risk of disease. The minor allele T of rs4845626 was associated with a reduced risk of lung cancer under all three models (p < 0.05). Similarly, allele C of rs4329505 had a protective effect on the risk of disease under the dominant and log-additive models (p < 0.05). In contrast, the genotype TT of rs892690 was associated with a 1.44-fold increased risk of lung cancer in a recessive model (p = 0.016). The minor alleles of rs745986 and rs2115819 were both associated with an elevated risk of disease in the dominant and log-additive models (p < 0.05). The allele A of rs2245121 also had a negative impact on the risk of disease in all three models (p < 0.05). Considering the differences in smoking status between the two groups, we conducted a stratification analysis (Table 5). The rs4845626 and rs4329505 polymorphisms of IL6R were associated with a reduced risk of lung cancer in both smokers and nonsmokers (p < 0.05). However, rs892690, rs745986 and rs2115819 of ALOX5 were associated with an increased risk of disease in nonsmokers, while rs2245121 of SFTPD was correlated with a risk of disease only in smokers (p < 0.05).
2.2. Genotyping Six candidate SNPs were selected based on the literature and the minor allele frequencies (MAF ≥ 5%) in Asian populations included in the 1000 Genomes database. DNA was extracted using a QIAamp DNA Blood Midi Kit (QIAGEN, Germany). Primers were designed using Sequenom MassARRAY Assay Design 3.0 software [19]. Genotypes were detected by a Sequenom MassARRAY RS1000 (Sequenom, San Diego, CA). 2.3. Statistical analysis Statistical analysis was performed with SPSS package version 20.0 (SPSS, Chicago, IL, USA). HaploReg v4.1 (https://pubs.broadinstitute. org/mammals/haploreg/haploreg.php) was used to predict the potential functions of the SNPs. MAFs of each SNP were checked for divergence from Hardy–Weinberg equilibrium (HWE). Allele and genotype frequencies in the cases and controls were evaluated using Chi-square tests. The effects of the SNPs on lung cancer risk were evaluated using logistic regression analysis and expressed by odds ratios (ORs) and 95% confidence intervals (CIs). Statistical significance was established when p < 0.05. 3. Results The demographic characteristics of the subjects are listed in Table 1. The case group consists of 302 males and 248 females, with a mean age of 58.2 years; the control group includes 300 males and 250 females, with a mean age of 58.5 years. No significant difference was observed in the distribution of sex, age, BMI or drinking status between the two groups (p > 0.05). However, the number of smokers in the case group was significantly greater than in the control group (p = 0.005). The lung cancer cases consist of 232 adenocarcinoma patients, 224 squamous cell carcinoma patients and 94 other types of lung cancer cases. The basic information for the candidate SNPs is described in Table 2. The predicted function according to the HaploReg database showed that the six SNPs were involved in regulation of the promoter and/or enhancer histone, changed motifs, DNAse, and GRASP and/or eQTL hits. All of the SNPs were consistent with HWE (p > 0.05). The differences in alleles and genotypes between the case and control groups are shown in Table 3. The major allele of each SNP was considered the reference allele, and the associations between minor alleles and the risk of lung cancer were evaluated. The allele T of rs4845626 was associated with a reduced risk of lung cancer
4. Discussion Beginning ten years ago, genome-wide association studies have provided several chromosomal regions, genes and SNPs that are associated with the risk of lung cancer [20–22]. Subsequently, these susceptibility polymorphisms have been proven to improve the accuracy of predicting the development of lung cancer to a considerable extent [23]. Currently, some specific SNPs have been applied to pharmacogenetic tests among patients with lung cancer [24]. Researchers have never stopped investigating gene polymorphisms that might be 2
International Immunopharmacology 79 (2020) 106155
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Table 2 Basic information about candidate SNPs of ALOX5, IL6R and SFTPD. SNP
Gene
Location
Position
Alleles
HaploReg
MAF-case
MAF-control
HWE p
rs4845626 rs4329505 rs892690 rs745986 rs2115819 rs2245121
IL6R IL6R ALOX5 ALOX5 ALOX5 SFTPD
Chr1 Chr1 Chr10 Chr10 Chr10 Chr10
154451009 154459944 45375240 45383460 45405641 79939482
G T C A A G
GRASP eQTL hits, Selected eQTL hits Enhancer histone marks, Motifs changed, GRASP eQTL hits, Selected eQTL hits Promoter and enhancer histone marks, DNAse, motifs changed, Selected eQTL hits Enhancer histone marks, DNAse, GRASP eQTL hits, Selected eQTL hits Enhancer histone marks, Motifs changed, Selected eQTL hits Promoter and enhancer histone marks, DNAse, motifs changed, GRASP eQTL hits, Selected eQTL hits
0.35 0.33 0.48 0.16 0.51 0.52
0.45 0.40 0.44 0.12 0.47 0.45
0.059 0.590 0.170 0.300 0.100 0.071
> > > > > >
T C T G G A
SNP: single nucleotide polymorphism, MAF: minor allele frequency, HWE: Hardy–Weinberg equilibrium.
rs3780894 in ALOX5 was associated with a risk of chronic rhinosinusitis [28]. Shen et al. demonstrated that carriers of the T allele of rs2115819 showed an increased risk of tuberculosis as children [29]. Habermann et al. showed that the interaction between ALOX15 rs11568131 and a high inflammation level was associated with an increased risk of colon cancer [30]. Jin et al. identified a significant correlation between rs2291427 in ALOX5 and the development of glioblastoma [31]. Our study showed that rs892690, rs745986 and rs2115819 in ALOX5 were all associated with an increased risk of lung cancer, consistent with the findings for ALOX5 polymorphisms in previous studies that they are involved in promoting inflammation in various diseases and tumors. We speculated that ALOX5 polymorphisms may also exert an influence on the development of lung cancer via their effects on the regulation of the synthesis of leukotriene. IL6 is an important cytokine with multiple functions, including regulating cell growth and differentiation, and it is involved in the immune response. The genetic polymorphisms of IL6 and/or IL6R are associated with respiratory and autoimmune diseases and several kinds of cancers. Wang et al. reported that the C allele of rs2228145 in IL6R
associated with the pathogenesis of lung cancer. The present study detected six SNPs in three inflammation-related genes, ALOX5, IL6R and SFTPD, in a case-control cohort and identified two SNPs (rs4845626 and rs4329505) associated with a decreased risk of lung cancer and four SNPs (rs892690, rs745986, rs2115819 and rs2245121) associated with an increased risk of the disease. The ALOX5 gene encodes lipoxygenase, which is a key enzyme in the process that catalyzes the conversion of arachidonic acid to leukotriene [25]. Moreover, leukotriene is closely associated with the process of inflammation and the development of several kinds of inflammatory disease [26]. Genetic variations in the promoter region of ALOX5 may lead to a reduced response to montelukast sodium (a kind of leukotriene antagonist) among patients with asthma, making it a very important pharmacogene [17]. In addition, polymorphisms of ALOX5 are also known to be associated with the risk of developing cardiovascular and respiratory diseases and a few types of cancers. Liu et al. found a strong linkage disequilibrium consisting of rs10900213, rs4293222 and rs2107545 in the ALOX5, ALOX5AP and MPO genes, and the haplotype exhibited a strong correlation with an increased risk of ischemic stroke [27]. Al-Shemari et al. reported that Table 3 Associations between alleles and genotypes of candidate SNPs and lung cancer risk. Gene
SNP
Allele/Genotype
Control (%)
Case (%)
OR (95% CI)
IL6R
rs4845626
G T GG GT TT T C TT TC CC
601 (54.6) 499 (45.4) 153 (27.8) 295 (53.6) 102 (18.6) 661 (60.1) 439 (39.9) 195 (35.5) 271 (49.3) 84 (15.3)
719 (65.4) 381 (34.6) 241 (43.8) 237 (43.1) 72 (13.1) 723 (66.8) 359 (33.2) 255 (47.1) 213 (39.4) 73 (13.5)
1 0.64 1 0.50 0.46 1 0.75 1 0.60 0.69
C T CC CT TT A G AA AG GG A G AA AG GG
614 (55.8) 486 (44.2) 163 (29.6) 288 (52.4) 99 (18.0) 972 (88.4) 128 (11.6) 432 (78.5) 108 (19.6) 10 (1.8%) 581 (52.8) 519 (47.2) 163 (29.6) 255 (46.4) 132 (24)
568 (51.9) 526 (48.1) 150 (27.4) 268 (49.0) 129 (23.6) 914 (84.5) 168 (15.6) 382 (70.6) 150 (27.7) 9 (1.7) 541 (49.2) 559 (50.8) 129 (23.4) 283 (51.5) 138 (25.1)
1 1.17 1 1.00 1.44 1 1.40 1 1.59 1.00 1 1.16 1.00 1.39 1.31
G A GG AG AA
609 491 179 251 120
527 567 133 261 153
1 1.33 (1.13–1.58) 1.00 1.40 (1.05–1.87) 1.75 (1.26–2.44)
rs4329505
ALOX5
rs892690
rs745986
rs2115819
SFTPD
rs2245121
(55.4) (44.6) (32.5) (45.6) (21.8)
OR: odds ratio, CI: confidence interval. *p < 0.05 indicates statistical significance. 3
(48.2) (51.8) (24.3) (47.7) (28.0)
p
(0.54–0.76)
< 0.001*
(0.39–0.66) (0.32–0.66)
< 0.001*
(0.63–0.90)
0.001*
(0.46–0.77) (0.48–1.00)
< 0.001*
(0.99–1.38)
0.067
(0.76–1.33) (1.02–2.04)
0.054
(1.09–1.79)
0.008*
(1.19–2.12) (0.40–2.52)
0.006
(0.98–1.37)
0.088
(1.04–1.86) (0.94–1.83)
0.076
0.001* 0.003*
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Table 4 Association between candidate SNPs and risk of lung cancer in three genetic models. Gene
SNP
Model
Genotype
Control
Case
OR (95% CI)
p
IL6R
rs4845626
Dominant
GG GT-TT GG-GT TT – TT TC-CC TT-TC CC –
153 (27.8) 397 (72.2) 448 (81.5) 102 (18.6) – 195 (35.5) 355 (64.5) 466 (84.7) 84 (15.3) –
241 (43.8) 309 (56.2) 478 (86.9) 72 (13.1) – 255 (47.1) 286 (52.9) 468 (86.5) 73 (13.5) –
1 0.49 1 0.68 0.64 1 0.62 1 0.90 0.76
< 0.001*
CC CT-TT CC-CT TT – AA AG-GG AA-AG GG – AA AG-GG AA-AG GG –
163 (29.6) 387 (70.4) 451 (82) 99 (18) – 432 (78.5) 118 (21.4) 540 (98.2) 10 (1.8) – 163 (29.6) 387 (70.4) 418 (76) 132 (24) –
150 (27.4) 397 (72.6) 418 (76.4) 129 (23.6) – 382 (70.6) 159 (29.4) 532 (98.3) 9 (1.7) – 129 (23.4) 421 (76.5) 412 (74.9) 138 (25.1) –
1 1.12 1 1.44 1.18 1 1.54 1 0.89 1.41 1 1.36 1 1.06 1.15
GG GA-AA GG-GA AA –
179 371 430 120 –
133 414 394 153 –
1 1.52 (1.16–1.98) 1 1.42 (1.08–1.88) 1.33 (1.12–1.57)
Recessive
rs4329505
Log-additive Dominant Recessive Log-additive
ALOX5
rs892690
Dominant Recessive
rs745986
Log-additive Dominant Recessive
rs2115819
Log-additive Dominant Recessive Log-additive
SFTPD
rs2245121
Dominant Recessive Log-additive
(32.5) (67.5) (78.2) (21.8)
(24.3) (75.7) (72) (28)
(0.38–0.63) 0.023* (0.49–0.95) (0.53–0.76)
< 0.001* < 0.001*
(0.48–0.79) 0.550 (0.64–1.27) (0.64–0.91)
0.002* 0.420
(0.86–1.45) 0.016* (1.07–1.94) (1.00–1.41)
0.054 0.002*
(1.17–2.04) 0.810 (0.36–2.24) (1.10–1.82)
0.007* 0.025*
(1.04–1.79) 0.700 (0.80–1.39) (0.97–1.36)
0.110 0.002* 0.013* < 0.001*
OR: odds ratio, CI: confidence interval. *p < 0.05 indicates statistical significance.
rs4845626 T and rs4329505 C exhibited a reduced risk of lung cancer, which may be attributable to different serum levels of IL6R. However, the detailed mechanism needs to be further explored. SFTPD is an important regulatory factor of the innate immune response in the human body. Moreover, SFTPD can protect lung function against the harms of inhaled hazardous substances [15]. Early studies have shown that the rs1923537 polymorphism of SFTPD is associated with postnatal pulmonary adaptation in preterm infants [35]. Subsequent study revealed the connection between the A allele of
was associated with the serum level of IL6R and decreased lung function in asthma patients [32]. Shoaib et al. demonstrated a correlation between rs2228145 and the risk of rheumatoid arthritis in a Pakistani population [33], while Zhang et al. observed a protective effect of rs2228145 in regards to the risk of developing gastric cancer in patients without Helicobacter pylori infection [34]. Our study focused on the polymorphisms rs4845626 and rs4329505, two SNPs previously associated with a decreased risk of COPD [16], in patients with lung cancer. We observed that carriers of
Table 5 Association of candidate SNPs with the risk of lung cancer stratified by smoking status. Gene
IL6R
SNP
rs4845626
rs4329505
ALOX5
rs892690
rs745986
rs2115819
SFTPD
rs2245121
Model
Smokers
Nonsmokers
OR (95% CI)
p
OR (95% CI)
p
Dominant Recessive Log-additive Dominant Recessive Log-additive
0.57 0.39 0.58 0.61 0.73 0.71
(0.39–0.85) (0.23–0.67) (0.44–0.77) (0.42–0.89) (0.41–1.27) (0.54–0.94)
0.006* < 0.001* < 0.001* 0.010* 0.260 0.014*
0.44 0.98 0.67 0.63 1.03 0.80
(0.32–0.62) (0.64–1.49) (0.54–0.85) (0.45–0.86) (0.67–1.58) (0.64–1.01)
< 0.001* 0.920 < 0.001* 0.004* 0.900 0.056
Dominant Recessive Log-additive Dominant Recessive Log-additive Dominant Recessive Log-additive
1.07 1.05 1.05 1.25 – 1.05 0.98 0.94 0.97
(0.71–1.62) (0.67–1.66) (0.80–1.37) (0.80–1.95)
0.740 0.820 0.730 0.320 – 0.830 0.910 0.790 0.820
1.15 1.81 1.29 1.76 2.62 1.70 1.63 1.09 1.25
(0.81–1.62) (1.22–2.68) (1.03–1.62) (1.23–2.52) (0.79–8.62) (1.23–2.35) (1.14–2.33) (0.75–1.58) (1.00–1.56)
0.440 0.003* 0.026* 0.002* 0.099 0.001* 0.007* 0.660 0.050
Dominant Recessive Log-additive
2.35 (1.55–3.57) 1.78 (1.13–2.81) 1.73 (1.32–2.27)
< 0.001* 0.011* < 0.001*
1.11 (0.79–1.57) 1.23 (0.86–1.76) 1.12 (0.90–1.38)
(0.70–1.57) (0.64–1.48) (0.62–1.44) (0.75–1.25)
4
0.550 0.260 0.300
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rs2243639 in SFTPD and the risk of COPD [36]. rs2245121 in SFTPD was also found to be correlated with an increased risk of COPD [18]. We observed an association between rs2245121 and the risk of lung cancer, suggesting that rs2245121 may affect the development of lung cancer by regulating the level of SFTPD and altering the innate immune response of patients. It is worth noting the stratification analysis results in our study. Two polymorphisms in IL6R were associated with a reduced risk of lung cancer in both subgroups, indicating that rs4845626 and rs4329505 are independent protective factors against the risk of disease. However, rs2245121 in SFTPD was correlated with an increased risk of disease only in smokers. As we mentioned above, SFTPD can protect the lungs against inhaled harmful substances, and thus, there may be an interaction between SFTPD polymorphisms and smoking that is involved in the development of lung cancer. This possibility needs to be confirmed in further studies. Although we identified six SNPs associated with an increased or decreased risk of lung cancer, the present study has two inevitable disadvantages. First, we were unable to collect information about the treatment regimens or the survival time of the patients, and therefore, we cannot evaluate the effect of these polymorphisms on the prognosis of the disease. Second, an association study can provide information about some specific correlations between SNPs and diseases, but not any details about the molecular mechanism. Future studies will focus on the correlation of these SNPs with the prognosis of the disease and the underlying mechanisms. In conclusion, we found that IL6R (rs4845626 and rs4329505) polymorphisms were associated with a decreased risk of lung cancer, and ALOX5 (rs892690, rs745986 and rs2115819) and SFTPD (rs2245121) polymorphisms were associated with an increased risk of the disease. SNP genotyping has been considered as an effective approach for lung cancer screening [37], and some biomedical companies have provided the service of early screening for lung cancer based on susceptible SNP genotyping [38]. Therefore, our results have provided candidate SNPs for early screening for lung cancer and new clues for further study of the pathogenesis of the disease.
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Yang, C. Zhu, H. Zhao, Y. Zhang, L. Chen, Y. Zhao, J. Li, Determinants of Gefitinib toxicity in advanced non-small cell lung cancer (NSCLC): a pharmacogenomic study of metabolic enzymes and transporters, Pharmacogenomics J. (2016). [25] Z. Elias, R. Paraskevi, S. Nikolaos, Variants of the arachidonate 5-lipoxygenaseactivating protein (ALOX5AP) gene and risk of stroke: a HuGE gene-disease association review and meta-analysis, Am. J. Epidemiol. 169 (5) (2009) 523–532. [26] G. Wang, Y. Wang, H. Sun, W. Cao, J. Zhang, H. Xiao, J. Zhang, Variants of the arachidonate 5-lipoxygenase-activating protein (ALOX5AP) gene and risk of ischemic stroke in Han Chinese of eastern China, J. Biomed. Res. 25 (5) (2011) 319–327. [27] D. Liu, L. Liu, Z. Song, Z. Hu, J. Liu, D. Hou, Genetic Variations of Oxidative Stress Related Genes ALOX5, ALOX5AP and MPO Modulate Ischemic Stroke Susceptibility Through Main Effects and Epistatic Interactions in a Chinese Population, Cell. Physiol. Biochem.: Int. J. Exp. Cell. Physiol., Biochem., Pharmacol. 43 (4) (2017) 1588–1602. [28] H. Al-Shemari, Y. Bossé, T.J. Hudson, M. Cabaluna, M. Duval, M. Lemire, S. ValleeSmedja, S. Frenkiel, M. Desrosiers, Influence of leukotriene gene polymorphisms on chronic rhinosinusitis, BMC Med. Genet. 9 (1) (2008) 21. [29] C. Shen, X.R. Wu, B.B. Wang, L. Sun, W.W. Jiao, J. Wang, W.X. Feng, J. Xiao, Q. Miao, F. Liu, Q.Q. Yin, X. Ma, A.D. Shen, ALOX5 is associated with tuberculosis in a subset of the pediatric population of North China, Genet. Test. Mol. Biomarkers 17 (4) (2013) 284–288. [30] N. Habermann, C.M. Ulrich, A. Lundgreen, K.W. Makar, E.M. Poole, B. Caan, R. Kulmacz, J. Whitton, R. Galbraith, J.D. Potter, PTGS1, PTGS2, ALOX5, ALOX12, ALOX15, and FLAP SNPs: interaction with fatty acids in colon cancer and rectal cancer, Genes Nutr. 8 (1) (2013) 115–126. [31] J. Tian-Bo, L. Xiao-Lan, Y. Hua, J. Mutu, S. Xu-Gang, Y. Dong-Ya, K. Long-Li, L. Shan-Qu, Association of polymorphisms in FLT3, EGFR, ALOX5, and NEIL3 with glioblastoma in the Han Chinese population, Med. Oncol. 30 (4) (2013) 718. [32] Y. Wang, H. Hu, J. Wu, X. Zhao, Y. Zhen, S. Wang, W. Li, M. Liang, B. Wu, G. Ma,
CRediT authorship contribution statement Xiaoping Wei: Conceptualization, Investigation, Writing - original draft. Chen Wang: Data curation, Investigation. Haiming Feng: Investigation. Bing Li: Investigation. Peng Jiang: Investigation. Jianbao Yang: Investigation. Duojie Zhu: Software. Shaobo Zhang: Validation. Tao Jin: Supervision. Yuqi Meng: . : Writing - review & editing. References [1] R.M. Feng, Y.N. Zong, S.M. Cao, R.H. Xu, Current cancer situation in China: good or bad news from the 2018 Global Cancer Statistics? Cancer Commun. 39 (1) (2019) 22. [2] Q.Y. Hong, G.M. Wu, G.S. Qian, C.P. Hu, J.Y. Zhou, L.A. Chen, W.M. Li, S.Y. Li, K. Wang, Q. Wang, Prevention and management of lung cancer in China, Cancer 121 (S17) (2015) 3080–3088. [3] T.-Y. Li, F. Zhang, Screening of lung cancer related SNPs and CNVs with SNP microarrays, Eur. Rev. Med. Pharmacol. Sci. 19 (2) (2015) 225–234. [4] K.A. Zanetti, Z. Wang, M. Aldrich, C.I. Amos, W.J. Blot, E.D. Bowman, L. Burdette, Q. Cai, N. Caporaso, C.C. Chung, Genome-wide association study confirms lung cancer susceptibility loci on chromosomes 5p15 and 15q25 in an African-American population, Lung Cancer 98 (2016) 33–42. [5] J.D. Mckay, R.J. Hung, Y. Han, X. Zong, R. Carrerastorres, D.C. Christiani, N.E. Caporaso, M. Johansson, X. Xiao, Y. Li, Large-scale association analysis identifies new lung cancer susceptibility loci and heterogeneity in genetic susceptibility across histological subtypes, Nat. Genet. 49 (7) (2017) 1126. [6] Z. Wang, W.J. Seow, K. Shiraishi, C.A. Hsiung, K. Matsuo, J. Liu, K. Chen, T. Yamji, Y. Yang, I.S. Chang, Meta-analysis of genome-wide association studies identifies multiple lung cancer susceptibility loci in never-smoking Asian women, Hum. Mol. Genet. 25 (3) (2016) 620–629. [7] D. Schottenfeld, J. Beebe-Dimmer, Chronic inflammation: a common and important factor in the pathogenesis of neoplasia, Ca Cancer J Clin 56 (2) (2010) 69–83. [8] Z. Jia, Z. Zhang, Q. Yang, C. Deng, D. Li, L. Ren, Effect of IL2RA and IL2RB gene
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human surfactant protein-D (SFTPD) gene and postnatal pulmonary adaptation in the preterm infant, Acta Paediat. 98 (1) (2009) 112–117. [36] M.S. Issac, W. Ashur, H. Mousa, Genetic polymorphisms of surfactant protein D rs2243639, Interleukin (IL)-1beta rs16944 and IL-1RN rs2234663 in chronic obstructive pulmonary disease, healthy smokers, and non-smokers, Mol. Diagnosis Therapy 18 (3) (2014) 343–354. [37] D.E. Midthun, Screening for Lung Cancer. Clin. Chest Med. 32(4) 659-668. [38] L. Mei, C. Huang, A. Wang, X. Zhang, Association between ADRB2, IL33, and IL2RB gene polymorphisms and lung cancer risk in a Chinese Han population, Int. Immunopharmacol. 77 (2019) 105930.
The IL6R gene polymorphisms are associated with sIL-6R, IgE and lung function in Chinese patients with asthma, Gene 585 (1) (2016) 51–57. [33] S. Ahmed, S. Hussain, A. Ammar, S. Jahan, S. Khaliq, H. Kaul, Interleukin 6 Receptor (IL6-R) Gene Polymorphisms Underlie Susceptibility to Rheumatoid Arthritis, Clin. Lab. 63 (9/2017) (2017) 1365–1369. [34] J.Z. Zhang, C.M. Liu, H.P. Peng, Y. Zhang, Association of genetic variations in IL-6/ IL-6R pathway genes with gastric cancer risk in a Chinese population, Gene 623 (2017) 1–4. [35] A. Hilgendorff, K. Heidinger, A.A. Kleinsteiber, I. Konig, A. Ziegler, U. Lindner, G. Frey, C. Merz, B. Lettgen, T. Chakraborty, Association of polymorphisms in the
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Effects of ALOX5, IL6R and SFTPD gene polymorphisms on the risk of lung cancer: A case-control study in China
T
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Xiaoping Wei , Chen Wang, Haiming Feng, Bing Li, Peng Jiang, Jianbao Yang, Duojie Zhu, Shaobo Zhang, Tao Jin, Yuqi Meng Department of Thoracic Surgery, Lanzhou University Second Hospital, Lanzhou, Gansu 730030, China
A R T I C LE I N FO
A B S T R A C T
Keywords: Lung cancer Polymorphisms Arachidonate 5-lipoxygenase (ALOX5) Interleukin 6 receptor (IL6R) Surfactant protein D (SFTPD)
Background: ALOX5, IL6R and SFTPD are all immune related genes that may be involved in the development of lung cancer. We sought to explore the effect of polymorphisms of these genes on the risk of lung cancer. Methods: Six single nucleotide polymorphisms (SNPs) were genotyped using a MassARRAY platform in a casecontrol cohort including 550 patients with lung cancer and 550 healthy controls. Results: The rs4845626-T and rs4329505-C alleles were associated with a decreased risk of lung cancer (p < 0.001), while the rs745986-G and rs2245121-A alleles were correlated with an increased risk of lung cancer (p < 0.01). The rs4845626-GT/GG and rs4329505-TC genotypes were protective against lung cancer (p < 0.001). However, the rs745986-AG and rs2245121-AG/AA genotypes were associated with an increased risk of lung cancer (p < 0.01). Stratification analysis showed that the rs4845626 and rs4329505 polymorphisms of IL6R were associated with a reduced risk of lung cancer in both smokers and nonsmokers (p < 0.05). However, rs892690, rs745986 and rs2115819 of ALOX5 were associated with an increased risk of disease in nonsmokers, while rs2245121 of SFTPD was correlated with a higher risk of disease in smokers (p < 0.05). Conclusion: Our results provide candidate SNPs for early screening for lung cancer and new clues for further study of the pathogenesis of the disease.
1. Introduction Lung cancer is a challenge for clinicians and researchers because of its high incidence and mortality, especially in developing countries, such as China [1]. Based on clinical experience, early detection and prevention of progression are crucial to prolong the survival time of patients with lung cancer [2]. In recent years, with the development of medical molecular biology, single nucleotide polymorphisms (SNPs) have been gradually found to have enormous potential in early screening and personalized treatment of lung cancer [3]. To date, a number of susceptibility genes and SNPs have been reported to have positive or negative correlations with the risk of developing lung cancer [4–6]. However, the current level of knowledge is insufficient for totally understanding the genetic background of lung cancer. Chronic inflammation is strongly associated with the proliferation, invasion and metastasis of tumor cells in the human body [7]. Moreover, polymorphisms in inflammation-related genes have been shown to be involved in the development of lung cancer [8,9]. These results drew our attention to the connection between polymorphisms in lung
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inflammation-related genes and the risk of lung cancer. Arachidonate 5lipoxygenase (ALOX5) is part of the leukotriene pathway and, therefore, plays important roles in the development of inflammation and inflammatory diseases [10]. ALOX5 polymorphisms have been associated with elevated levels of leukotriene, resulting in impairment of lung function [11]. However, few studies have directly investigated the correlation between ALOX5 polymorphisms and lung cancer risk. Interleukin 6 (IL6) polymorphisms have been intensively investigated in patients with lung cancer in several different populations [12–14], but the correlation between IL6 receptor (IL6R) polymorphisms and risk of disease is still unclear. Surfactant protein D (SFTPD) regulates inflammatory processes and protects the host against pathogens invading pulmonary alveolar type II cells, indicating it has a potential role in blocking the progression of lung cancer [15]. Therefore, the present study selected ALOX5, IL6R and SFTPD as candidate genes to investigate. rs4845626 and rs4329505 in IL6R have been associated with a decreased risk of COPD in Mexicans [16]. rs892690, rs745986 and
Corresponding author at: #82 Cuiyingmen, Lanzhou, Gansu 730030, China. E-mail addresses: [email protected], [email protected] (X. Wei).
https://doi.org/10.1016/j.intimp.2019.106155 Received 16 October 2019; Received in revised form 24 December 2019; Accepted 25 December 2019 1567-5769/ © 2020 Elsevier B.V. All rights reserved.
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rs2115819 in ALOX5 were shown to be associated with the metabolism of montelukast in patients with asthma [17]. Moreover, rs2245121 in SFTPD was found to be correlated with an increased risk of COPD [18]. In the present study, we sought to explore the association between these SNPs and the risk of lung cancer.
Table 1 The demographic characteristics of the participants. Variables
Sex (%) Male Female Age (mean ± SD), years BMI (mean ± SD), kg/ m2 Smoking (%) Ever Never Drinking (%) Ever Never Histology (%) Adenocarcinoma Squamous Other
2. Materials and methods 2.1. Participants All subjects are ethnic Chinese Han living in Lanzhou city or the surrounding areas. A total of 550 lung cancer patients and 550 healthy individuals were recruited for the study as a case and a control group, respectively. The cases were diagnosed with lung cancer by pathological biopsy at our hospital. Cases with any other types of cancers, immune diseases or mental disorders were excluded. The controls were recruited at the physical examination center of our hospital and included only those with no history of cancer, lung disease, or immune disease. We obtained written informed consent from each subject, and the study was approved by the Ethics Department of Lanzhou University Second Hospital.
Case (%) (n = 550)
Control (%) (n = 550)
302 (54.9) 248 (45.1) 58.2 ± 9.7
300 (54.5) 250 (45.5) 58.5 ± 10.2
24.3 ± 3.5
24.5 ± 3.8
255 (46.4) 295 (53.6)
209 (38.0) 341 (62.0)
224 (40.7) 326 (59.3)
232 (42.2) 318 (57.8)
χ 2/t
p
0.015
0.903
0.838
0.186
0.564
0.242
7.887
0.005*
0.240
0.624
232 (42.2) 224 (40.7) 94 (17.1)
(OR = 0.64, 95% CI = 0.54–0.76, p < 0.001). The allele C of rs4329505 also exhibited a protective effect against the risk of lung cancer (OR = 0.75, 95% CI = 0.63–0.90, p = 0.001). In contrast, the allele G of rs745986 was correlated with a 1.40-fold increased risk of lung cancer (95% CI = 1.09–1.79, p = 0.008), and the allele A of rs2245121 was also associated with an increased risk of disease (OR = 1.33, 95% CI = 1.13–1.58, p = 0.001). For the genotypes, both the GT and GG genotypes of rs4845626 were correlated with a decreased risk of lung cancer (p < 0.001). The TC genotype of rs4329505 also showed a protective effect against the risk of lung cancer (p < 0.001). However, the AG genotype of rs745986 (p = 0.006) and the AG and AA genotypes of rs2245121 (p = 0.003) were all associated with an increased risk of lung cancer. The associations between genotypes and the risk of lung cancer were also evaluated based on genetic models (Table 4). All of the SNPs were identified as having associations with the risk of disease. The minor allele T of rs4845626 was associated with a reduced risk of lung cancer under all three models (p < 0.05). Similarly, allele C of rs4329505 had a protective effect on the risk of disease under the dominant and log-additive models (p < 0.05). In contrast, the genotype TT of rs892690 was associated with a 1.44-fold increased risk of lung cancer in a recessive model (p = 0.016). The minor alleles of rs745986 and rs2115819 were both associated with an elevated risk of disease in the dominant and log-additive models (p < 0.05). The allele A of rs2245121 also had a negative impact on the risk of disease in all three models (p < 0.05). Considering the differences in smoking status between the two groups, we conducted a stratification analysis (Table 5). The rs4845626 and rs4329505 polymorphisms of IL6R were associated with a reduced risk of lung cancer in both smokers and nonsmokers (p < 0.05). However, rs892690, rs745986 and rs2115819 of ALOX5 were associated with an increased risk of disease in nonsmokers, while rs2245121 of SFTPD was correlated with a risk of disease only in smokers (p < 0.05).
2.2. Genotyping Six candidate SNPs were selected based on the literature and the minor allele frequencies (MAF ≥ 5%) in Asian populations included in the 1000 Genomes database. DNA was extracted using a QIAamp DNA Blood Midi Kit (QIAGEN, Germany). Primers were designed using Sequenom MassARRAY Assay Design 3.0 software [19]. Genotypes were detected by a Sequenom MassARRAY RS1000 (Sequenom, San Diego, CA). 2.3. Statistical analysis Statistical analysis was performed with SPSS package version 20.0 (SPSS, Chicago, IL, USA). HaploReg v4.1 (https://pubs.broadinstitute. org/mammals/haploreg/haploreg.php) was used to predict the potential functions of the SNPs. MAFs of each SNP were checked for divergence from Hardy–Weinberg equilibrium (HWE). Allele and genotype frequencies in the cases and controls were evaluated using Chi-square tests. The effects of the SNPs on lung cancer risk were evaluated using logistic regression analysis and expressed by odds ratios (ORs) and 95% confidence intervals (CIs). Statistical significance was established when p < 0.05. 3. Results The demographic characteristics of the subjects are listed in Table 1. The case group consists of 302 males and 248 females, with a mean age of 58.2 years; the control group includes 300 males and 250 females, with a mean age of 58.5 years. No significant difference was observed in the distribution of sex, age, BMI or drinking status between the two groups (p > 0.05). However, the number of smokers in the case group was significantly greater than in the control group (p = 0.005). The lung cancer cases consist of 232 adenocarcinoma patients, 224 squamous cell carcinoma patients and 94 other types of lung cancer cases. The basic information for the candidate SNPs is described in Table 2. The predicted function according to the HaploReg database showed that the six SNPs were involved in regulation of the promoter and/or enhancer histone, changed motifs, DNAse, and GRASP and/or eQTL hits. All of the SNPs were consistent with HWE (p > 0.05). The differences in alleles and genotypes between the case and control groups are shown in Table 3. The major allele of each SNP was considered the reference allele, and the associations between minor alleles and the risk of lung cancer were evaluated. The allele T of rs4845626 was associated with a reduced risk of lung cancer
4. Discussion Beginning ten years ago, genome-wide association studies have provided several chromosomal regions, genes and SNPs that are associated with the risk of lung cancer [20–22]. Subsequently, these susceptibility polymorphisms have been proven to improve the accuracy of predicting the development of lung cancer to a considerable extent [23]. Currently, some specific SNPs have been applied to pharmacogenetic tests among patients with lung cancer [24]. Researchers have never stopped investigating gene polymorphisms that might be 2
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Table 2 Basic information about candidate SNPs of ALOX5, IL6R and SFTPD. SNP
Gene
Location
Position
Alleles
HaploReg
MAF-case
MAF-control
HWE p
rs4845626 rs4329505 rs892690 rs745986 rs2115819 rs2245121
IL6R IL6R ALOX5 ALOX5 ALOX5 SFTPD
Chr1 Chr1 Chr10 Chr10 Chr10 Chr10
154451009 154459944 45375240 45383460 45405641 79939482
G T C A A G
GRASP eQTL hits, Selected eQTL hits Enhancer histone marks, Motifs changed, GRASP eQTL hits, Selected eQTL hits Promoter and enhancer histone marks, DNAse, motifs changed, Selected eQTL hits Enhancer histone marks, DNAse, GRASP eQTL hits, Selected eQTL hits Enhancer histone marks, Motifs changed, Selected eQTL hits Promoter and enhancer histone marks, DNAse, motifs changed, GRASP eQTL hits, Selected eQTL hits
0.35 0.33 0.48 0.16 0.51 0.52
0.45 0.40 0.44 0.12 0.47 0.45
0.059 0.590 0.170 0.300 0.100 0.071
> > > > > >
T C T G G A
SNP: single nucleotide polymorphism, MAF: minor allele frequency, HWE: Hardy–Weinberg equilibrium.
rs3780894 in ALOX5 was associated with a risk of chronic rhinosinusitis [28]. Shen et al. demonstrated that carriers of the T allele of rs2115819 showed an increased risk of tuberculosis as children [29]. Habermann et al. showed that the interaction between ALOX15 rs11568131 and a high inflammation level was associated with an increased risk of colon cancer [30]. Jin et al. identified a significant correlation between rs2291427 in ALOX5 and the development of glioblastoma [31]. Our study showed that rs892690, rs745986 and rs2115819 in ALOX5 were all associated with an increased risk of lung cancer, consistent with the findings for ALOX5 polymorphisms in previous studies that they are involved in promoting inflammation in various diseases and tumors. We speculated that ALOX5 polymorphisms may also exert an influence on the development of lung cancer via their effects on the regulation of the synthesis of leukotriene. IL6 is an important cytokine with multiple functions, including regulating cell growth and differentiation, and it is involved in the immune response. The genetic polymorphisms of IL6 and/or IL6R are associated with respiratory and autoimmune diseases and several kinds of cancers. Wang et al. reported that the C allele of rs2228145 in IL6R
associated with the pathogenesis of lung cancer. The present study detected six SNPs in three inflammation-related genes, ALOX5, IL6R and SFTPD, in a case-control cohort and identified two SNPs (rs4845626 and rs4329505) associated with a decreased risk of lung cancer and four SNPs (rs892690, rs745986, rs2115819 and rs2245121) associated with an increased risk of the disease. The ALOX5 gene encodes lipoxygenase, which is a key enzyme in the process that catalyzes the conversion of arachidonic acid to leukotriene [25]. Moreover, leukotriene is closely associated with the process of inflammation and the development of several kinds of inflammatory disease [26]. Genetic variations in the promoter region of ALOX5 may lead to a reduced response to montelukast sodium (a kind of leukotriene antagonist) among patients with asthma, making it a very important pharmacogene [17]. In addition, polymorphisms of ALOX5 are also known to be associated with the risk of developing cardiovascular and respiratory diseases and a few types of cancers. Liu et al. found a strong linkage disequilibrium consisting of rs10900213, rs4293222 and rs2107545 in the ALOX5, ALOX5AP and MPO genes, and the haplotype exhibited a strong correlation with an increased risk of ischemic stroke [27]. Al-Shemari et al. reported that Table 3 Associations between alleles and genotypes of candidate SNPs and lung cancer risk. Gene
SNP
Allele/Genotype
Control (%)
Case (%)
OR (95% CI)
IL6R
rs4845626
G T GG GT TT T C TT TC CC
601 (54.6) 499 (45.4) 153 (27.8) 295 (53.6) 102 (18.6) 661 (60.1) 439 (39.9) 195 (35.5) 271 (49.3) 84 (15.3)
719 (65.4) 381 (34.6) 241 (43.8) 237 (43.1) 72 (13.1) 723 (66.8) 359 (33.2) 255 (47.1) 213 (39.4) 73 (13.5)
1 0.64 1 0.50 0.46 1 0.75 1 0.60 0.69
C T CC CT TT A G AA AG GG A G AA AG GG
614 (55.8) 486 (44.2) 163 (29.6) 288 (52.4) 99 (18.0) 972 (88.4) 128 (11.6) 432 (78.5) 108 (19.6) 10 (1.8%) 581 (52.8) 519 (47.2) 163 (29.6) 255 (46.4) 132 (24)
568 (51.9) 526 (48.1) 150 (27.4) 268 (49.0) 129 (23.6) 914 (84.5) 168 (15.6) 382 (70.6) 150 (27.7) 9 (1.7) 541 (49.2) 559 (50.8) 129 (23.4) 283 (51.5) 138 (25.1)
1 1.17 1 1.00 1.44 1 1.40 1 1.59 1.00 1 1.16 1.00 1.39 1.31
G A GG AG AA
609 491 179 251 120
527 567 133 261 153
1 1.33 (1.13–1.58) 1.00 1.40 (1.05–1.87) 1.75 (1.26–2.44)
rs4329505
ALOX5
rs892690
rs745986
rs2115819
SFTPD
rs2245121
(55.4) (44.6) (32.5) (45.6) (21.8)
OR: odds ratio, CI: confidence interval. *p < 0.05 indicates statistical significance. 3
(48.2) (51.8) (24.3) (47.7) (28.0)
p
(0.54–0.76)
< 0.001*
(0.39–0.66) (0.32–0.66)
< 0.001*
(0.63–0.90)
0.001*
(0.46–0.77) (0.48–1.00)
< 0.001*
(0.99–1.38)
0.067
(0.76–1.33) (1.02–2.04)
0.054
(1.09–1.79)
0.008*
(1.19–2.12) (0.40–2.52)
0.006
(0.98–1.37)
0.088
(1.04–1.86) (0.94–1.83)
0.076
0.001* 0.003*
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Table 4 Association between candidate SNPs and risk of lung cancer in three genetic models. Gene
SNP
Model
Genotype
Control
Case
OR (95% CI)
p
IL6R
rs4845626
Dominant
GG GT-TT GG-GT TT – TT TC-CC TT-TC CC –
153 (27.8) 397 (72.2) 448 (81.5) 102 (18.6) – 195 (35.5) 355 (64.5) 466 (84.7) 84 (15.3) –
241 (43.8) 309 (56.2) 478 (86.9) 72 (13.1) – 255 (47.1) 286 (52.9) 468 (86.5) 73 (13.5) –
1 0.49 1 0.68 0.64 1 0.62 1 0.90 0.76
< 0.001*
CC CT-TT CC-CT TT – AA AG-GG AA-AG GG – AA AG-GG AA-AG GG –
163 (29.6) 387 (70.4) 451 (82) 99 (18) – 432 (78.5) 118 (21.4) 540 (98.2) 10 (1.8) – 163 (29.6) 387 (70.4) 418 (76) 132 (24) –
150 (27.4) 397 (72.6) 418 (76.4) 129 (23.6) – 382 (70.6) 159 (29.4) 532 (98.3) 9 (1.7) – 129 (23.4) 421 (76.5) 412 (74.9) 138 (25.1) –
1 1.12 1 1.44 1.18 1 1.54 1 0.89 1.41 1 1.36 1 1.06 1.15
GG GA-AA GG-GA AA –
179 371 430 120 –
133 414 394 153 –
1 1.52 (1.16–1.98) 1 1.42 (1.08–1.88) 1.33 (1.12–1.57)
Recessive
rs4329505
Log-additive Dominant Recessive Log-additive
ALOX5
rs892690
Dominant Recessive
rs745986
Log-additive Dominant Recessive
rs2115819
Log-additive Dominant Recessive Log-additive
SFTPD
rs2245121
Dominant Recessive Log-additive
(32.5) (67.5) (78.2) (21.8)
(24.3) (75.7) (72) (28)
(0.38–0.63) 0.023* (0.49–0.95) (0.53–0.76)
< 0.001* < 0.001*
(0.48–0.79) 0.550 (0.64–1.27) (0.64–0.91)
0.002* 0.420
(0.86–1.45) 0.016* (1.07–1.94) (1.00–1.41)
0.054 0.002*
(1.17–2.04) 0.810 (0.36–2.24) (1.10–1.82)
0.007* 0.025*
(1.04–1.79) 0.700 (0.80–1.39) (0.97–1.36)
0.110 0.002* 0.013* < 0.001*
OR: odds ratio, CI: confidence interval. *p < 0.05 indicates statistical significance.
rs4845626 T and rs4329505 C exhibited a reduced risk of lung cancer, which may be attributable to different serum levels of IL6R. However, the detailed mechanism needs to be further explored. SFTPD is an important regulatory factor of the innate immune response in the human body. Moreover, SFTPD can protect lung function against the harms of inhaled hazardous substances [15]. Early studies have shown that the rs1923537 polymorphism of SFTPD is associated with postnatal pulmonary adaptation in preterm infants [35]. Subsequent study revealed the connection between the A allele of
was associated with the serum level of IL6R and decreased lung function in asthma patients [32]. Shoaib et al. demonstrated a correlation between rs2228145 and the risk of rheumatoid arthritis in a Pakistani population [33], while Zhang et al. observed a protective effect of rs2228145 in regards to the risk of developing gastric cancer in patients without Helicobacter pylori infection [34]. Our study focused on the polymorphisms rs4845626 and rs4329505, two SNPs previously associated with a decreased risk of COPD [16], in patients with lung cancer. We observed that carriers of
Table 5 Association of candidate SNPs with the risk of lung cancer stratified by smoking status. Gene
IL6R
SNP
rs4845626
rs4329505
ALOX5
rs892690
rs745986
rs2115819
SFTPD
rs2245121
Model
Smokers
Nonsmokers
OR (95% CI)
p
OR (95% CI)
p
Dominant Recessive Log-additive Dominant Recessive Log-additive
0.57 0.39 0.58 0.61 0.73 0.71
(0.39–0.85) (0.23–0.67) (0.44–0.77) (0.42–0.89) (0.41–1.27) (0.54–0.94)
0.006* < 0.001* < 0.001* 0.010* 0.260 0.014*
0.44 0.98 0.67 0.63 1.03 0.80
(0.32–0.62) (0.64–1.49) (0.54–0.85) (0.45–0.86) (0.67–1.58) (0.64–1.01)
< 0.001* 0.920 < 0.001* 0.004* 0.900 0.056
Dominant Recessive Log-additive Dominant Recessive Log-additive Dominant Recessive Log-additive
1.07 1.05 1.05 1.25 – 1.05 0.98 0.94 0.97
(0.71–1.62) (0.67–1.66) (0.80–1.37) (0.80–1.95)
0.740 0.820 0.730 0.320 – 0.830 0.910 0.790 0.820
1.15 1.81 1.29 1.76 2.62 1.70 1.63 1.09 1.25
(0.81–1.62) (1.22–2.68) (1.03–1.62) (1.23–2.52) (0.79–8.62) (1.23–2.35) (1.14–2.33) (0.75–1.58) (1.00–1.56)
0.440 0.003* 0.026* 0.002* 0.099 0.001* 0.007* 0.660 0.050
Dominant Recessive Log-additive
2.35 (1.55–3.57) 1.78 (1.13–2.81) 1.73 (1.32–2.27)
< 0.001* 0.011* < 0.001*
1.11 (0.79–1.57) 1.23 (0.86–1.76) 1.12 (0.90–1.38)
(0.70–1.57) (0.64–1.48) (0.62–1.44) (0.75–1.25)
4
0.550 0.260 0.300
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rs2243639 in SFTPD and the risk of COPD [36]. rs2245121 in SFTPD was also found to be correlated with an increased risk of COPD [18]. We observed an association between rs2245121 and the risk of lung cancer, suggesting that rs2245121 may affect the development of lung cancer by regulating the level of SFTPD and altering the innate immune response of patients. It is worth noting the stratification analysis results in our study. Two polymorphisms in IL6R were associated with a reduced risk of lung cancer in both subgroups, indicating that rs4845626 and rs4329505 are independent protective factors against the risk of disease. However, rs2245121 in SFTPD was correlated with an increased risk of disease only in smokers. As we mentioned above, SFTPD can protect the lungs against inhaled harmful substances, and thus, there may be an interaction between SFTPD polymorphisms and smoking that is involved in the development of lung cancer. This possibility needs to be confirmed in further studies. Although we identified six SNPs associated with an increased or decreased risk of lung cancer, the present study has two inevitable disadvantages. First, we were unable to collect information about the treatment regimens or the survival time of the patients, and therefore, we cannot evaluate the effect of these polymorphisms on the prognosis of the disease. Second, an association study can provide information about some specific correlations between SNPs and diseases, but not any details about the molecular mechanism. Future studies will focus on the correlation of these SNPs with the prognosis of the disease and the underlying mechanisms. In conclusion, we found that IL6R (rs4845626 and rs4329505) polymorphisms were associated with a decreased risk of lung cancer, and ALOX5 (rs892690, rs745986 and rs2115819) and SFTPD (rs2245121) polymorphisms were associated with an increased risk of the disease. SNP genotyping has been considered as an effective approach for lung cancer screening [37], and some biomedical companies have provided the service of early screening for lung cancer based on susceptible SNP genotyping [38]. Therefore, our results have provided candidate SNPs for early screening for lung cancer and new clues for further study of the pathogenesis of the disease.
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