- Open Access
Association between interleukin 8–251 T/A and +781 C/T polymorphisms and glioma risk
Diagnostic Pathology volume 10, Article number: 138 (2015)
Gliomas are aggressive tumors of the central nervous system that rely on production of growth factors for tumor progression. Interleukin 8 (IL-8) is up-regulated in gliomas to promote angiogenesis and proliferation. The aim of this study was to evaluate the association of the IL-8 -251 T/A and +781 C/T polymorphisms and glioma risk.
We enrolled 300 glioma patients and 300 age- and gender-matched healthy controls. A prospective hospital-based case–control design and logistic regression analysis were utilized. The IL-8 gene polymorphisms were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).
Glioma patients had a significantly higher frequency of IL-8 -251 AA genotype [odds ratio (OR) =1.91, 95 % confidence interval (CI) = 1.22, 3.00; P = 0.005] and IL-8 -251 A allele (OR =1.36, 95 % CI = 1.08, 1.70; P = 0.009) than controls. When stratified by the grade of glioma, patients with WHO IV glioma had a significantly higher frequency of IL-8 -251 AA genotype (OR =1.56, 95 % CI = 1.01, 2.39; P = 0.04).
To the best of our knowledge, this is the first report in the literature that the IL-8 -251 AA genotype and A allele were at a higher risk for glioma.
Gliomas make up about 30 % of all brain and central nervous system tumors and 80 % of all malignant brain tumors . Despite the growing number of preclinical and clinical trials focused on the treatment of malignant gliomas, the prognosis for this disease remains grim [2–4]. Only rare familial syndromes and exposure to high therapeutic doses of ionizing radiation are known causes of glioma . Some other factors are also found to affect glioma risk, such as: high levels of processed meat consumption and obesity during adolescence [6, 7]. Recent research has focused on identifying germline polymorphisms associated with risk of glioma, and using molecular markers to classify glial tumors into more homogenous groups [5, 8, 9].
Interleukin-8 (IL-8), which is best known for its leukocyte chemotactic properties and associated role in inflammatory and infectious diseases , is now known to possess tumorigenic and proangiogenic properties as well [11, 12]. IL-8 is encoded by the IL-8 gene located on chromosome 4q13-21, consisting of four exons, three introns, and the proximal promoter region . Several SNPs have been reported in the IL-8 gene and some of them, such as −251 T/A (rs4073) and +781 C/T (rs2227306), can regulate the IL-8 production [14–16]. In human gliomas, IL-8 is expressed and secreted at high levels both in vitro and in vivo, and recent experiments suggest it is critical to glial tumor neovascularity and progression .
We hypothesized that common genetic variants in IL-8 gene influenced the risk of glioma. To test this hypothesis, we performed a prospective hospital-based case–control study to evaluate the association of the IL-8 -251 T/A and +781 C/T polymorphisms and glioma risk.
In this prospective hospital-based case–control study, we enrolled 300 glioma patients and 300 age- and gender-matched healthy controls between May 2012 and May 2014 in the Department of Neurosurgery, The First Affiliated Hospital of Xi'an Jiao Tong University, China. Tumor type and stage were determined according to the WHO criteria . In addition, similar to the cases the controls were all required to be born in China to native Chinese Han parents. To confirm the diagnosis, two physicians reviewed the hospital records and validated each case. Collected clinical data included sex, age, mean education, smoking, alcohol drinking and family history of cancer. All data points were collected through interviews with the patient or their families/surrogates. All parts of the study were approved by the Institutional Ethical Committee of the First Affiliated Hospital of Xi'an Jiao Tong University, and informed consent according to the Declaration of Helsinki was obtained from all participants or their families/surrogates.
DNA extraction and genotyping
Genomic DNA was isolated from white blood cells by the commercially available Qiagen kit (QIAGEN Inc., Valencia, CA, USA). IL-8 -251 A/T and +781 C/T polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) as previous described [18–20]. Restriction enzyme and primer sequences of IL-8 promoter SNPs were listed in Table 1. The PCR cycling conditions were 5 min at 94 °C followed by 35 cycles of 1 min at 94 °C, 1 min at 61 °C, and 2 min at 72 °C, with a final step at 72 °C for 20 min. The PCR product was subjected to digestion for 4 h with restriction enzyme at 37 °C. Electrophoresis in a 2.5 % agarose gel followed by ethidium bromide staining and ultraviolet illumination allowed detection of the alleles. For quality control, two independent observers, read all genotypes without knowing about the case or control status. When replicate quality control samples were evaluated, genotypes showed 100 % concordance.
Data are presented as means ± standard deviation (SD) or as percentages for categorical variables. Differences between continuous variables were assessed by Student’s t test, while those between categorical variables were evaluated using Pearson x 2 test. The existence of differences in genotypic frequencies between groups were assessed by means of Pearson x 2 test and calculating the odds ratio (OR) with the 95 % confidence intervals (CI). Statistical significance was taken at nominal P-value < 0.05 for all comparisons. SAS version 9.1 (SAS Institute, Cary, NC) was used for all statistical tests.
Characteristics of participants
Characteristics of glioma cases and healthy controls were presented in Table 2. No significant differences were found between the glioma cases and healthy controls in sex, age, mean education, smoking, alcohol drinking and family history of cancer (Table 2). Among 300 glioma cases, 223 cases had astrocytomas, 43 cases had ependymomas, 19 cases had oligodendrogliomas, and 15 cases had mixed gliomas (Table 2). In these cases, 17 were WHO grade I gliomas, 106 were WHO grade II gliomas, 76 were WHO grade III gliomas, and 101 were WHO grade IV gliomas (Table 2).
IL-8 -251 T/A polymorphisms and glioma risk
Glioma patients had a significantly higher frequency of IL-8 -251 AA genotype (OR =1.91, 95 % CI = 1.22, 3.00; P = 0.005) and IL-8 -251 A allele (OR =1.36, 95 % CI = 1.08, 1.70; P = 0.009) than controls (Table 3). When stratified by the grade of glioma, patients with WHO IV glioma had a significantly higher frequency of IL-8 -251 AA genotype (OR =1.56, 95 % CI = 1.01, 2.39; P = 0.04) (Table 4). When stratified by the histology of glioma, there was no significant difference in the distribution of each genotype (Table 4).
IL-8 + 781 C/T polymorphisms and glioma risk
No association was found between IL-8 + 781 C/T polymorphisms and glioma risk (Table 3).
Recent research has focused on identifying germline polymorphisms associated with risk of glioma. A study by Zong et al. suggested that the Cyclin D1 gene G870A polymorphism was a risk factor for the development of glioma . There was a potential decreased susceptibility to glioma in association with the XRCC1 Arg399Gln polymorphism, especially in Asians . Regulator of telomere elongation helicase 1 (RTEL1) rs6010620 polymorphism was likely to be associated with increased glioma risk . A study by Yuan et al. suggested that common variants in ERCC1 may contribute to susceptibility to glioma, especially in Asians . The AA genotype of ERCC1 C8092A might be associated with a higher risk of adult glioma than the CA and CC genotypes and that the risk allele of ERCC2 K751Q confered a significant susceptibility to adult glioma, especially in Asian populations . Strong evidence for the association between XRCC3 C18607T polymorphism and glioma risk was found in a meta-analysis . There was a significant association between EGF +61A > G polymorphism and glioma risk among Asians . The polymorphism of interleukin-4 receptor alpha (IL-4Ralpha) rs1801275 played a protective role in the glioma pathogenesis, particularly among Asians .
The IL-8 gene polymorphisms were also associated with many other cancers. A recent prospective case–control study found that IL-8 polymorphism was associated with ovarian cancer . The IL-8 -251 AA genotype was at a higher risk for oral cancer in the Caucasian population [30, 31]. The IL-8-251 TA or AA genotype conferred risk of cardia gastric cancer in a population in Southwestern China . The IL-8 -251 AA genotype was associated with the overall risk of developing gastric cancer and may seem to be more susceptible to overall gastric cancer in Asian populations . The IL-8 -251 T/A polymorphism was associated with lung cancer susceptibility in Asians and the −251 A allele may increase risk of lung cancer in Asians . A HuGE review and meta-analysis based on 42 case–control studies found that -251A allele was susceptible in the development of low-penetrance cancers . Two case–control studies found that IL-8 -251 T/A polymorphism was significantly associated with colorectal cancer susceptibility risk [36, 37]. The IL-8-251 T/A polymorphism was associated with breast cancer risk . A study found that IL-8 -251 T/A polymorphism was associated with bladder cancer susceptibility and outcome after bacillus Calmette-Guerin immunotherapy in a northern Indian cohort .
The mechanisms of the IL-8 -251 AA genotype and A allele as risk factors of glioma are still unclear. In human gliomas, IL-8 is expressed and secreted at high levels both in vitro and in vivo, and recent experiments suggest it is critical to glial tumor neovascularity and progression . There is strong evidence that IL-8 secretion is associated with glioma formation and malignant progression . The IL-8 -251A allele in a homozygous state has been associated with increased expression of the IL-8 gene [40, 41]. It is entirely plausible that the the IL-8 -251 AA genotype and A allele will affect the risk of gliomas.
There is limitation that needs to be acknowledged and addressed regarding the present study. First of all, these results should be interpreted with caution because the population was only from China, which reduces the possibility of confounding from ethnicity, so it does not permit extrapolation of the results to other ethnic groups. Second, this study is limited by its size and lack of replication. Additional large scale studies are needed to confirm this finding. Third, since controls were recruited from those who came to hospitals for routine health examination, there was a certain risk of selection bias. Finally, rare familial syndromes and exposure to high therapeutic doses of ionizing radiation are known causes of glioma, which were not explored in the present study.
To the best of our knowledge, this is the first report in the literature that the IL-8 -251 AA genotype and A allele were at a higher risk for glioma. Additional large scale studies are needed to confirm this finding. Genetic risk profiling has the potential to identify individuals who have not yet had glioma develop but are at high risk. Analysis of polymorphic variants will potentially provide a tool to assess the risk of glioma decades before diagnosis. This process provides ample opportunity to implement behavioral and environmental changes.
All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
Informed consent was obtained from all individual participants included in the study.
Goodenberger ML, Jenkins RB. Genetics of adult glioma. Cancer Genet. 2012;205:613–21.
Ohgaki H, Kleihues P. Epidemiology and etiology of gliomas. Acta Neuropathol. 2005;109:93–108.
Jakola AS, Myrmel KS, Kloster R, Torp SH, Lindal S, Unsgard G, et al. Comparison of a strategy favoring early surgical resection vs a strategy favoring watchful waiting in low-grade gliomas. JAMA. 2012;308:1881–8.
Murphy KA, Erickson JR, Johnson CS, Seiler CE, Bedi J, Hu P, et al. CD8+ T cell-independent tumor regression induced by Fc-OX40L and therapeutic vaccination in a mouse model of glioma. J Immunol. 2014;192:224–33.
Schwartzbaum JA, Fisher JL, Aldape KD, Wrensch M. Epidemiology and molecular pathology of glioma. Nat Clin Pract Neurol. 2006;2:494–503. quiz 491 p following 516.
Wei Y, Zou D, Cao D, Xie P. Association between processed meat and red meat consumption and risk for glioma: A meta-analysis from 14 articles. Nutrition. 2015;31:45–50.
Moore SC, Rajaraman P, Dubrow R, Darefsky AS, Koebnick C, Hollenbeck A, et al. Height, body mass index, and physical activity in relation to glioma risk. Cancer Res. 2009;69:8349–55.
Shete S, Hosking FJ, Robertson LB, Dobbins SE, Sanson M, Malmer B, et al. Genome-wide association study identifies five susceptibility loci for glioma. Nat Genet. 2009;41:899–904.
Wrensch M, Jenkins RB, Chang JS, Yeh RF, Xiao Y, Decker PA, et al. Variants in the CDKN2B and RTEL1 regions are associated with high-grade glioma susceptibility. Nat Genet. 2009;41:905–8.
Harada A, Sekido N, Akahoshi T, Wada T, Mukaida N, Matsushima K. Essential involvement of interleukin-8 (IL-8) in acute inflammation. J Leukoc Biol. 1994;56:559–64.
Xie K. Interleukin-8 and human cancer biology. Cytokine Growth Factor Rev. 2001;12:375–91.
Brat DJ, Bellail AC, Van Meir EG. The role of interleukin-8 and its receptors in gliomagenesis and tumoral angiogenesis. Neuro Oncol. 2005;7:122–33.
Mukaida N, Shiroo M, Matsushima K. Genomic structure of the human monocyte-derived neutrophil chemotactic factor IL-8. J Immunol. 1989;143:1366–71.
Fey MF, Tobler A. An interleukin-8 (IL-8) cDNA clone identifies a frequent HindIII polymorphism. Hum Genet. 1993;91:298.
Hacking D, Knight JC, Rockett K, Brown H, Frampton J, Kwiatkowski DP, et al. Increased in vivo transcription of an IL-8 haplotype associated with respiratory syncytial virus disease-susceptibility. Genes Immun. 2004;5:274–82.
Ohyauchi M, Imatani A, Yonechi M, Asano N, Miura A, Iijima K, et al. The polymorphism interleukin 8–251 A/T influences the susceptibility of Helicobacter pylori related gastric diseases in the Japanese population. Gut. 2005;54:330–5.
Louis DN, Ohgaki H, Wiestler OD, Cavenee WK, Burger PC, Jouvet A, et al. The 2007 WHO classification of tumours of the central nervous system. Acta Neuropathol. 2007;114:97–109.
Arinir U, Klein W, Rohde G, Stemmler S, Epplen JT, Schultze-Werninghaus G. Polymorphisms in the interleukin-8 gene in patients with chronic obstructive pulmonary disease. Electrophoresis. 2005;26:2888–91.
Qin X, Deng Y, Liao XC, Mo CJ, Li X, Wu HL, et al. The IL-8 gene polymorphisms and the risk of the hepatitis B virus/infected patients. DNA Cell Biol. 2012;31:1125–30.
Olivo-Diaz A, Romero-Valdovinos M, Gudino-Ramirez A, Reyes-Gordillo J, Jimenez-Gonzalez DE, Ramirez-Miranda ME, et al. Findings related to IL-8 and IL-10 gene polymorphisms in a Mexican patient population with irritable bowel syndrome infected with Blastocystis. Parasitol Res. 2012;111:487–91.
Zong H, Cao L, Ma C, Zhao J, Ming X, Shang M, et al. Association between the G870A polymorphism of Cyclin D1 gene and glioma risk. Tumour Biol. 2014;35:8095–101.
Zhu W, Yao J, Li Y, Xu B. Assessment of the association between XRCC1 Arg399Gln polymorphism and glioma susceptibility. Tumour Biol. 2014;35:3061–6.
Zhao W, Bian Y, Zhu W, Zou P, Tang G. Regulator of telomere elongation helicase 1 (RTEL1) rs6010620 polymorphism contribute to increased risk of glioma. Tumour Biol. 2014;35:5259–66.
Yuan G, Gao D, Ding S, Tan J. DNA repair gene ERCC1 polymorphisms may contribute to the risk of glioma. Tumour Biol. 2014;35:4267–75.
Xu Z, Ma W, Gao L, Xing B. Association between ERCC1 C8092A and ERCC2 K751Q polymorphisms and risk of adult glioma: a meta-analysis. Tumour Biol. 2014;35:3211–21.
Wang R, Li M, Gao WW, Gu Y, Guo Y, Wang G, et al. Quantitative assessment of the association between XRCC3 C18607T polymorphism and glioma risk. Tumour Biol. 2014;35:1101–5.
Tao Y, Liang G. Quantitative assessment of the association between +61A > G polymorphism of epidermal growth factor gene and susceptibility to glioma. Tumour Biol. 2014;35:369–77.
Guo J, Shi L, Li M, Xu J, Yan S, Zhang C, et al. Association of the interleukin-4Ralpha rs1801275 and rs1805015 polymorphisms with glioma risk. Tumour Biol. 2014;35:573–9.
Koensgen D, Bruennert D, Ungureanu S, Sofroni D, Braicu EI, Sehouli J, et al. Polymorphism of the IL-8 gene and the risk of ovarian cancer. Cytokine. 2015;71:334–8.
Yang L, Zhu X, Liang X, Ling Z, Li R. Association of IL-8-251A > T polymorphisms with oral cancer risk: evidences from a meta-analysis. Tumour Biol. 2014;35:9211–8.
Wang Z, Wang C, Zhao Z, Liu F, Guan X, Lin X, et al. Association between -251A > T polymorphism in the interleukin-8 gene and oral cancer risk: a meta-analysis. Gene. 2013;522:168–76.
Pan XF, Wen Y, Loh M, Wen YY, Yang SJ, Zhao ZM, et al. Interleukin-4 and −8 gene polymorphisms and risk of gastric cancer in a population in Southwestern China. Asian Pac J Cancer Prev. 2014;15:2951–7.
Xue H, Liu J, Lin B, Wang Z, Sun J, Huang G. A meta-analysis of interleukin-8 -251 promoter polymorphism associated with gastric cancer risk. PLoS One. 2012;7, e28083.
Gao P, Zhao H, You J, Jing F, Hu Y. Association between interleukin-8 -251A/T polymorphism and risk of lung cancer: a meta-analysis. Cancer Invest. 2014;32:518–25.
Wang N, Zhou R, Wang C, Guo X, Chen Z, Yang S, et al. −251 T/A polymorphism of the interleukin-8 gene and cancer risk: a HuGE review and meta-analysis based on 42 case–control studies. Mol Biol Rep. 2012;39:2831–41.
Mustapha MA, Shahpudin SN, Aziz AA, Ankathil R. Risk modification of colorectal cancer susceptibility by interleukin-8 -251 T > A polymorphism in Malaysians. World J Gastroenterol. 2012;18:2668–73.
Walczak A, Przybylowska K, Dziki L, Sygut A, Chojnacki C, Chojnacki J, et al. The lL-8 and IL-13 gene polymorphisms in inflammatory bowel disease and colorectal cancer. DNA Cell Biol. 2012;31:1431–8.
Huang Q, Wang C, Qiu LJ, Shao F, Yu JH. IL-8-251A > T polymorphism is associated with breast cancer risk: a meta-analysis. J Cancer Res Clin Oncol. 2011;137:1147–50.
Ahirwar DK, Mandhani A, Mittal RD. IL-8 -251 T > A polymorphism is associated with bladder cancer susceptibility and outcome after BCG immunotherapy in a northern Indian cohort. Arch Med Res. 2010;41:97–103.
Taguchi A, Ohmiya N, Shirai K, Mabuchi N, Itoh A, Hirooka Y, et al. Interleukin-8 promoter polymorphism increases the risk of atrophic gastritis and gastric cancer in Japan. Cancer Epidemiol Biomarkers Prev. 2005;14:2487–93.
Hull J, Thomson A, Kwiatkowski D. Association of respiratory syncytial virus bronchiolitis with the interleukin 8 gene region in UK families. Thorax. 2000;55:1023–7.
Thanks are expressed to all coinvestigators, local project coordinators, research assistants, laboratory technicians, and secretaries/administrative assistants.
All authors declare that they have no competing interests.
HL and PM carried out the molecular genetic studies and drafted the manuscript. CX and WX carried out the genotyping. MW and HJ participated in the design of the study and performed the statistical analysis. HL, PM, CX, WX, MW and HJ conceived of the study, and participated in its design and coordination and helped to draft the manuscript. All authors read and approved the final manuscript.
Rights and permissions
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
About this article
Cite this article
Liu, H., Mao, P., Xie, C. et al. Association between interleukin 8–251 T/A and +781 C/T polymorphisms and glioma risk. Diagn Pathol 10, 138 (2015). https://doi.org/10.1186/s13000-015-0378-x
- Glioma Patient
- Glioma Risk
- XRCC1 Arg399Gln
- Glioma Case
- XRCC1 Arg399Gln Polymorphism