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10.1245/s10434-006-9122-9
Annals of Surgical Oncology 14:329-339 (2007)
© 2007 Society of Surgical Oncology
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Original Article

Host Cytokine Genotype is Related to Adverse Prognosis and Systemic Inflammation in Gastro-Oesophageal Cancer

Chris Deans, MBChB1, Matthew Rose-Zerilli, BSc2, Stephen Wigmore, MD1, James Ross, PhD1, Martin Howell, PhD3, Alan Jackson, MD4, Robert Grimble, PhD4 and Kenneth Fearon, MD1,5

1 Cell Injury and Apoptosis Section, Tissue Injury and Repair Group, MRC Centre for Inflammation Research, Department of Clinical and Surgical Sciences, Medical School, Edinburgh University, Edinburgh, EH8 9AG, UK
2 Histocompatibility and Immunogenetics Laboratory, Human Genetics Division, University of Southampton, Southampton, UK
3 Department of Histocompatibility and Immunogenetics, National Blood Service, Holland Drive, Newcastle-upon-Tyne, NE2 4NQ, UK
4 Institute of Human Nutrition, School of Medicine, University of Southampton, SO16 7PX, Southampton, UK
5 University Department of Surgery, Royal Infirmary, 51 Little France Crescent, Edinburgh, EH16 4SA, UK

Correspondence: Address correspondence and reprint requests to: Kenneth Fearon, MD; E-mail: K.Fearon{at}ed.ac.uk


   ABSTRACT
TOP
 ABSTRACT
INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Background: Systemic inflammation has been linked with reduced survival in cancer, however, the role of the host cytokine genotype versus tumour phenotype in the generation of this response is not clearly established. This study examined the relationship between cytokine polymorphisms (IL-1ß 511, IL-6 174, IL-10 1082, TNF{alpha} 308 and LT{alpha} +252) and serum cytokine concentrations, serum CRP concentration and survival duration in patients with gastro-oesophageal malignancy.

Methods: Two hundred and three newly diagnosed patients with gastric or oesophageal cancer had serum CRP and cytokine concentrations determined by ELISA. SNP genotyping was performed by Taqman allelic discrimination genotyping and compared with the genotype observed in 266 healthy volunteers. Clinico-pathological information was collected prospectively and survival duration was recorded.

Results: Distribution of the cytokine genotypes was similar between patients and controls. The IL-6 174 CC and IL-10 1082 GG genotypes were associated with elevated serum CRP (P = .03, P = .01, respectively; Mann–Whitney U test) and sTNF-R (P = .015, P = .02) concentrations. These genotypes were also associated with reduced survival duration (P = .01, P = .047; log-rank test). TNF{alpha} AA genotype was also associated with reduced survival duration on univariate (P = .032) and multivariate analysis (P = .006, multivariate model), but not with inflammatory markers. No other cytokine polymorphisms were associated with systemic inflammatory markers or prognosis.

Conclusions: There is a pro-inflammatory cytokine haplotype (IL-6 CC, IL-10 GG, TNF{alpha} AA) that is associated with adverse prognosis that may act, at least in part, through an inflammatory mediated mechanism. Determining patients’ cytokine haplotype may improve prognostication and allow stratification for intervention studies.

Key Words: Cytokines • Polymorphisms • Cancer • Survival • Inflammation


   INTRODUCTION
TOP
 ABSTRACT
 INTRODUCTION
MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Malignant tumours arising around the oesophagogastric junction have the fastest increase in incidence of any solid tumour in Europe and North America.1,2 Together gastric and oesophageal malignancy are the third leading cause of cancer-related death in the UK.3 Despite advances in both staging techniques and treatments, 5-year-survival rarely exceeds 30%.4 Systemic inflammation has been associated with a number of malignant diseases, including gastric and oesophageal cancer, and is associated with adverse outcome.5–15 Among patients with gastric cancer the presence of systemic inflammation has been associated with a markedly reduced median survival (53 vs. 9 weeks, P < .001).14 Similarly, a recent Japanese study has identified a shortened survival in oesophageal cancer patients with an elevated serum C-reactive protein (CRP) at the time of diagnosis.15 The systemic inflammatory response is modulated by the complex interaction of many pro- and anti-inflammatory cytokines. Although the production of these mediators is partly responsive to environmental influences, the origin of systemic inflammation among cancer patients remains obscure and may be influenced by host cytokine genotype. Variations in genotype for a number of cytokines have been associated with changes in both serum cytokine and serum acute phase protein concentrations and prognosis among cancer patients.

Tumour necrosis factor {alpha} (TNF{alpha}), interleukin-1ß (IL-1ß), and IL-6 are key pro-inflammatory cytokines involved in the generation of the inflammatory response. Increased production of these cytokines has been shown to contribute to adverse outcome in patients with sepsis, infections and inflammatory diseases, such as inflammatory bowel disease and rheumatoid arthritis.16–19 Our group has previously investigated the influence of IL-1ß polymorphisms on systemic inflammation and survival in patients with pancreatic cancer.20 The possession of the less common allele 2 was associated with increased levels of serum CRP concentrations, increased production of IL-1ß by peripheral blood mononuclear cells (PBMC), and reduced overall survival. In contrast, another group failed to identify an association between IL-1ß polymorphisms and outcome for patients with ovarian cancer.21

A single nucleotide polymorphism (SNP) at the 308 position of the TNF{alpha} gene lies within the promoter region and the A allele has been associated with higher levels of TNF{alpha} production in vitro following stimulation with endotoxin.22 Variation at the 308 locus has been associated with conflicting levels of TNF{alpha} production both in in vitro studies and among patients with sepsis.23,24 Our own work, again on pancreatic cancer patients, found no link between genotype and serum soluble TNF receptor (sTNF-R) concentrations or serum CRP concentrations.25 There was a trend for reduced survival duration associated with possession of the AA genotype, but this did not reach statistical significance (P = .13). Another group identified that possession of the A allele was associated with elevated serum sTNF-R concentrations in patients with non-Hodgkin’s lymphoma and was associated with reduced overall survival.26

A SNP at the 252 locus of the lymphotoxin {alpha} (LT{alpha}) (tumour necrosis factor ß) gene has also been shown to modify levels of TNF production. Septic surgical patients homozygous for the AA genotype had significantly higher mortality than those patients who were homozygous for GG.27 Among cancer patients, homozygotes had a more favourable prognosis than heterozygotes in advanced lung cancer and patients possessing the type 1 allele were shown to have poorer survival in oesophageal cancer.28,29

A SNP at 174 in the IL-6 promoter region has been associated with increased serum IL-6 concentrations and worse outcome among cardiac surgery patients.30 Survival data for cancer patients is conflicting. The possession of the C allele has been associated with earlier stage disease and better outcome (independent of stage) in women with ovarian cancer and breast cancer.31,32 However, the same genotype has also been associated with adverse survival in another group of patients with breast cancer.33

Interleukin-10 (IL-10) is generally regarded as an anti-inflammatory cytokine important in the attenuation of the inflammatory response. The IL-10 gene polymorphism at position 1082 lies within the promoter region and the AA genotype is generally thought to be associated with reduced levels of IL-10 production, but this is contradicted by one study.34,35 The high IL-10 producer genotype (GG) has been linked with more advanced stage among patients with gastric cancer.36

With a view to understanding more clearly the role of host genotype in the genesis of tumour-related systemic inflammation this study examined the relationship between cytokine polymorphisms (IL-1ß 511, IL-6 174, IL-10 1082, TNF{alpha} 308 and LT{alpha} +252) and serum cytokine concentrations, serum CRP concentrations and survival duration among newly diagnosed patients with gastro-oesophageal cancer.


   MATERIALS AND METHODS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
RESULTS
 DISCUSSION
 REFERENCES
 
Study Patients and Controls
Patients diagnosed with gastric or oesophageal cancer between March 2002 and May 2004 within Lothian and Borders regions were invited to take part in the study. All patients provided written informed consent and the study received ethical permission from the Lothian Research Ethics Committee. Histological confirmation of disease was established in all cases following endoscopic biopsy. Patients were staged with a combination of computerised tomography (CT), endoscopic ultrasound (EUS) and laparoscopy/laparoscopic ultrasound (LUS) according to the International Union Against Cancer (UICC).37 Final histopathological stage (pTNM) was only available for those patients who underwent surgical resection. In all other cases the final clinical stage (cTNM), as agreed at the unit multidisciplinary team meeting (MDT), was recorded. Tumours around the oesophagogastric junction were classified according to Siewert and those classified as type I and II were staged as oesophageal and type III were staged as gastric.38 For the purposes of analysis the cTNM stage was used for all patients. Separate subgroup analysis of those patients with final histopathological (pTNM) stage gave identical results. Clinical and pathological information was recorded prospectively for each patient, including treatment modality, tumour grade and histological subtype. Duration of survival, defined as time from histological diagnosis to death, was recorded for all patients.

In addition to the patient group, blood samples were also collected from healthy controls for genotyping. The control group used in this study comprised 266 British Caucasian bone marrow and solid organ donors, collected via the Histocompatibility and Immunogenetics Laboratory, Southampton University Hospitals. The mean age of these controls (140 males, 126 females) was 39.2 years at the time of blood collection (age range 3–69). All cytokine genotyping results in this control group have been published previously.39,40

Serum Cytokine and CRP Measurement
Whole blood was collected from each patient at the time of diagnosis. Samples were collected from patients without evidence of infection and at least 2 weeks following any invasive investigation to avoid artificial induction of an acute phase systemic response. CRP was determined using an automated immunoturbidimetric assay (Abbott TDX, Abbott Laboratories, Maidenhead, UK). A level above 10 mg/l was considered evidence of an acute phase response.

Cytokine concentrations were determined by sandwich enzyme-linked immunosorbent assay (ELISA) using module kits and performed according to the manufacturers instructions (Caltag, Bender MedSystems, Towcester, UK) as described previously.41 The lower limit of sensitivity for each assay was: <1 pg/ml IL-1ß, 1.4 pg/ml IL-6, .8 pg/ml IL-10 and 5.8 pg/ml sTNF-R.

Cytokine Genotyping
Genomic DNA was extracted from samples of lithium-heparinised blood using the Wizard Genomic DNA purification kit (Promega, Southampton, UK). The following SNPs were selected for genotyping due to their documented but variable association with cytokine production: IL-1ß 511, IL-6 174, IL-10 1082, TNF{alpha} 308 and LT{alpha} +252. Genotyping was carried out by TaqMan allelic discrimination genotyping on the 7900HT Sequence Detection System (Applied Biosystems, Warrington, UK). Primers and TaqMan probes were designed using Primer Express version 2.0 software (sequences shown in Table 1Go) and synthesised and supplied by Applied Biosystems UK. Ten microlitre PCR reactions containing 20 ng of DNA, .9 and .2 µM probes (final concentrations) were performed in 384 well plates. Each genotyping plate contained no DNA Template (water) controls; SDS version 2.1 software was used to analyse real-time and end-point fluorescence. Around 50 samples (~25% of the sample group) were randomly selected and included as replicates for each genotype tested. All replicates agreed. Only between 3–8 samples failed to genotype for each SNP (96.1–98.5% completion rate).


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  		TABLE 1. Cytokine TaqMan genotyping primer and probe sequences
 
Statistical Analysis
Comparisons between groups of continuous variables were made by the Mann–Whitney U test. Categorical variables were compared by the Chi-squared test. Correlations between continuous variables were assessed by the Spearman rank correlation coefficient. Survival between groups was analysed by the log-rank test and Cox’s proportional hazard’s model was used for multivariate analysis. A P value ≤ .05 was considered statistically significant. Significant P values were corrected for multiple comparisons using Bonferroni correction.


   RESULTS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
DISCUSSION
 REFERENCES
 
Study Patients and Genotype Distributions
Patient demographics are outlined in Table 2Go. Two hundred and three patients were genotyped. The median age was 71 years (interquartile range 62–78) and 66% of patients were male. The primary tumour sites were oesophageal (n = 91, 45%), gastric (n = 75, 37%) and those arising from the gastro-oesophageal junction (OGJ) (n = 37, 18%). Histological confirmation of disease was obtained in all cases and the predominant histological subtype was adenocarcinoma (85%). 113 (56%) patients underwent surgical resection, 22 of these received pre-operative chemotherapy. Seven (3%) patients, all of whom had squamous cell carcinoma of the oesophagus, received chemo-irradiation with curative intent. The remaining patients (n = 90) were not suitable for curative therapy and received palliative chemo/radiotherapy or underwent alternative palliative treatments, such as insertion of a stent or endoscopic laser therapy.


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  		TABLE 2. Study patient demographics (n = 203)
 
The prevalence of cytokine genotypes are presented in Table 3Go. There was a significant difference between cases and controls with the IL-1ß 511 cc [P = .01, OR = 1.664 (95% CI = 1.137–2.434)] and ct [P = .03, OR = .657 (95% CI = .453–.955)] genotypes, but it did not remain so when corrected for multiple comparisons, making this finding most likely occurring due to chance. There were no significant differences between cancer patients and healthy controls in the distribution of any of the other genotypes. All genotype frequencies in both the patient and control groups were distributed in accordance with the Hardy–Weinberg equilibrium except for the IL-6 patient group (P = .05), the TNF{alpha} control group (P = .04) and the LT{alpha} control group (P = .002).


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  		TABLE 3. Allelic distribution of cytokine genotypes among the study population
 
Serum Cytokine and CRP Concentrations
The median serum concentration of IL-6 was 9.1 pg/ml (interquartile range 0–88 pg/ml) and for sTNF-R the median concentration was 3.3 ng/ml (interquartile range 1.9–5.8 ng/ml). IL-1ß and IL-10 were only detectable in 4 (2%) and 10 (5%) patient’s serum, respectively. 82 (41%) patients had an elevated acute phase protein response (CRP > 10 mg/l).

Serum sTNF-R concentration correlated with serum CRP concentration (r = .38, P < .001; spearman rank test), but there was no significant relationship between the other serum cytokine concentrations and CRP level.

Link between Genotype and Mediators/Markers of Systemic Inflammation
The IL-6 CC genotype was associated with elevated serum sTNF-R concentrations when compared with the GC genotype (P = .015; Mann–Whitney U test), but not when compared with the GG genotype (P = .14). Similarly, the IL-10 GG genotype was also associated with elevated sTNF-R concentrations when compared with the AG genotype (P = .05), but not when compared with the AA allele (P = .22). There was no association between any other serum cytokine concentrations and IL-6 or IL-10 genotypes. In addition, there was no association between any IL-1ß, TNF{alpha} or LT{alpha} genotypes and serum cytokine concentrations.

The IL-6 CC homozygous status was associated with significantly elevated serum CRP concentrations when compared with GC and GG genotypes [median 13 mg/l (range 4–35 mg/l) versus median 6 mg/l (range 2–22 mg/l)] (P = .03, Mann–Whitney U test) the GG IL-10 genotype was also associated with elevated serum CRP levels [median 16 mg/l (range 3–34 mg/l)] when compared with the AA genotype [median 6 mg/l (range 3–13 mg/l)] (P = .02, Mann–Whitney U test) and the AG genotype [median 5 mg/l (range 2–24 mg/l)] (P = .03). The GG homozygous genotype was associated with elevated serum CRP levels (P = .01). None of the other cytokine genotypes were associated with serum CRP concentrations.

Possession of one or more of the following genotypes, IL-6 CC, IL-10 GG or TNF{alpha} AA, was associated with increasing serum CRP concentrations (P = .013, Chi square test) (Fig. 1Go). Moreover, multivariate analysis demonstrated that this positive association was independent of both treatment modality undertaken and stage of disease [regression coefficient = .16 (95% CI .05–.64), P = .021; linear regression analysis].


Figure 1
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  		FIG. 1. Possession of one or more of the following genotypes, IL-6 CC, IL-10 GG or TNF{alpha} AA, was associated with increasing serum CRP concentrations (P = .013, Chi squared test).

 
Survival Analysis
Survival of patients by genotype is presented in Figs. 2Go and 3Go. Possession of the CC genotype for IL-6 was associated with reduced survival duration when compared with GG or GC genotype (median survival 256 vs. 431 days) (P = .010; log-rank test) (Fig. 2AGo). The homozygous genotype GG for IL-10 was associated with reduced survival compared with AA/AG genotypes (median survival 310 vs. 389 days, P = .047) (Fig. 2BGo). Similarly, AA homozygosity for the TNF{alpha} genotype was associated with adverse prognosis (median survival 194 vs. 409 days, P = .032) (Fig. 2CGo). Furthermore, the possession of more than one of these genotypes was associated with reduced survival (no alleles, median survival 512 days; 1 allele, median survival 269 days; 2 or 3 alleles, median survival 258 days; P = .004) (Fig. 3Go). Genotypes for IL-1ß and LT{alpha} were not associated with prognosis.


Figure 2
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  		FIG. 2. Kaplan–Meier survival plots presented by (A) interleukin-6 (IL-6) genotype [CC median survival 256 days (heavy line), GC/GG median survival 431 days (light line)] (P = .010; Log-rank test); (B) interleukin-10 (IL-10) genotype [GG median survival 310 days (heavy line), AA/AG median survival 389 days (light line)] (P = .047) and (C) tumour necrosis factor {alpha} (TNF{alpha}) genotype [AA median survival 194 days (heavy line), AG/GG median survival 409 days (light line)] (P = .032).

 

Figure 3
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  		FIG. 3. Kaplan–Meier survival plot stratified for possession of interleukin-6 (IL-6) CC genotype, interleukin-10 (IL-10) GG genotype and tumour necrosis factor {alpha} (TNF{alpha}) AA genotype. Possession of no alleles (light line), median survival 512 days; 1 allele (medium line), median survival 269 days; 2 or 3 alleles (heavy line), median survival 258 days (P = .004; Log-rank test).

 
Multivariate testing identified the AA genotype for TNF{alpha} as an independent adverse prognostic indicator when tested with age, sex, stage of disease, grade of tumour and serum CRP concentration (P = .006, hazard ratio 2.5; Cox’s proportional hazard’s model) (Table 4Go). Increasing age, male sex, advanced disease stage, elevated serum CRP concentration, and the TNF{alpha} AA genotype were all independently associated with poor prognosis. Genotypes for IL-6 and IL-10 were not independent prognostic indicators (P = .48 and P = .18, respectively).


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  		TABLE 4. Multivariate survival analysis of 203 patients with gastro-oesophageal cancer using Cox/s proportional hazard’s model
 
Eighty-two (41%) patients had an elevated acute phase protein response (CRP > 10 mg/l) at the time of diagnosis and this was associated with a significantly reduced survival duration; CRP < 10 mg/l median survival 550 days versus CRP > 10 mg/l median survival 217 days (P < .001; log-rank test) (Fig. 4Go). Moreover, multivariate testing also identified CRP as an independent prognostic indicator (P = .002, hazard ratio 1.1; Cox’s proportional hazard’s model) (Table 4Go).


Figure 4
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  		FIG. 4. Kaplan–Meier survival plot according to the presence or absence of systemic inflammation. CRP < 10 mg/l median survival = 550 days (light line) versus CRP > 10 mg/l median survival 217 days (heavy line) (P < .001; Log-rank test).

 

   DISCUSSION
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
REFERENCES
 
In the present study the IL-6 174 CC and IL-10 1082 GG genotypes were associated with elevated markers of systemic inflammation (CRP and sTNF-R concentrations) and were associated with reduced survival duration. In addition, the TNF{alpha} 308 AA genotype was not associated with systemic inflammation, but was identified as an independent adverse prognostic marker. Therefore, there is a pro-inflammatory cytokine haplotype that is associated with adverse prognosis among patients with gastro-oesophageal cancer that may act in association with, but not entirely with, an inflammatory mediated mechanism.

IL-1ß and IL-10 are not routinely measurable in the serum of healthy controls or cancer patients. In this study IL-1ß and IL-10 were only detectable in 4 (2%) and 10 (5%) patient’s serum, respectively. We, therefore, did not find an association between genotype and serum concentrations of these cytokines. We have, however, previously shown an association between IL-1ß genotype and IL-1ß production by PBMC in pancreatic cancer patients, but not with serum IL-1ß concentrations.20 PBMC production of IL-1ß may better reflect tissue IL-1ß levels rather than circulating serum concentrations and perhaps is a more accurate determinant of IL-1ß activity.

TNF{alpha} is also rarely detected in the serum, however, soluble TNF receptors (sTNF-R) are shed in response to TNF{alpha} release and may be used as an indirect measure of TNF{alpha} concentration.42 Serum sTNF-R concentrations were higher among those patients with elevated serum CRP concentrations. We found a significant correlation between serum sTNF-R and serum CRP concentrations (r = .38, P < .001) and we have shown a similar association among patients with pancreatic cancer.43 There was no association between TNF genotype and serum sTNF-R concentrations either in this study or our previous work.25 The data relating TNF{alpha} production and genotype appears contradictory and at present there is no clear evidence relating TNF{alpha} genotype to circulating TNF{alpha} levels. However, tissue levels of TNF{alpha} may be a more relevant measure of TNF activity rather than systemic concentrations, which was not measured in the present study.

Forty-one percent of patients had an elevated acute phase response (CRP > 10 mg/l). Systemic inflammation has been found in association with the majority of solid epithelial malignancies and around 50% of patients may have an acute phase response (APPR) at the time of diagnosis.43 The presence of an elevated CRP has been associated with adverse prognosis in a number of types of cancer, independent of stage of disease.5–15 We have similarly identified an elevated serum CRP concentration as an adverse prognostic indicator among patients with gastro-oesophageal cancer, also independent of stage of disease. The presence of systemic inflammation in malignant disease is an important marker of tumour behaviour and clearly has clinical relevance in assisting management decision making as well as emphasising the therapeutic potential of targeted anti-inflammatory strategies in advanced cancer.

IL-6 and IL-10 genotypes were associated with serum CRP concentrations in this study. The IL-6 CC genotype was associated with elevated serum concentrations of CRP and sTNF-R. A study of healthy volunteers identified the presence of the 174C allele to be associated with higher baseline CRP levels.44 In cancer patients the rates of production of IL-6 can be linked to markers of systemic inflammation such as CRP. Although our study did not identify any association between IL-6 genotype and serum IL-6 concentrations, it is possible that the CC genotype may be associated with elevated IL-6 production, which acts at the tissue level to promote an acute phase response. The similar association between IL-6 genotype and sTNF-R may reflect the more stable nature of the receptor molecule compared with the other cytokines, and in this regard sTNF-R may behave more as a marker of systemic inflammation.

We also found the GG IL-10 genotype to be associated with elevated serum CRP and sTNF-R concentrations. The 1082 AA polymorphism is generally thought to be associated with reduced levels of IL-10 production.34 One would therefore expect the GG genotype to be associated with increased levels of IL-10 production. An association between increased levels of an anti-inflammatory cytokine and elevated concentrations of acute phase proteins may initially appear contradictory, but may simply reflect the increased counter-regulatory activity of this important anti-inflammatory mediator in response to the presence of systemic inflammation.

We found no association between IL-1ß, TNF{alpha} or LT{alpha} genotypes and CRP concentrations. In pancreatic cancer patients we previously demonstrated an association between allele 2 IL-1ß genotype and elevated serum CRP levels, but in the present study there was no such association between the SNP at position 511 and CRP levels.20 Similarly, there was no association between TNF{alpha} polymorphisms and CRP, a finding also in support of our previous work on pancreatic cancer patients.25 Data relating to the 308 polymorphism and levels of TNF{alpha} production remain contradictory, however polymorphisms at this locus do not appear to influence serum CRP levels. Serum CRP concentrations were measured in a group of patients who had undergone cardiac surgery and this study did not find any differences between CRP levels and TNF{alpha} 308 genotypes.45 Another group similarly failed to demonstrate differences in CRP concentrations by 308 genotype among smokers.46

The IL-6 174 CC and the IL-10 1082 GG genotypes were associated with reduced survival duration. The association between systemic inflammation and adverse prognosis among cancer patients has been well documented and in this study we have shown these two genotypes to be associated with elevated acute phase protein (CRP) concentrations. It is therefore possible that the adverse prognosis associated with these polymorphisms is related to the presence of systemic inflammation. This is supported by multivariate analysis where these genotypes lost their significance as prognostic indicators when CRP was co-analysed. The CC IL-6 genotype has been linked with adverse prognosis among breast cancer patients, where possession of the CC polymorphism was associated with higher grade tumours and worse overall survival.33 The GG genotype for IL-10 1082 has previously been associated with advanced stage in gastric cancer patients and associated reduced survival.36

We also found the AA TNF{alpha} 308 genotype to be related to adverse prognosis. In contrast to the IL-6 and IL-10 genotypes, we found no association between TNF{alpha} polymorphisms and systemic inflammation. It is therefore less likely that the reduced survival associated with this cytokine is related entirely to the generation of an inflammatory response. In addition, the AA genotype was an independent prognostic indicator on multivariate analysis, independent of CRP concentration, stage and other clinico-pathological characteristics that have previously been associated with adverse prognosis. We previously identified an association between the AA genotype and reduced survival in pancreatic cancer patients and another group similarly found the possession of the A allele to be linked with adverse outcome in patients with non-Hodgkin’s lymphoma.25,26 However, the AA genotype was only identified in 8% of patients, making it less useful in the clinical setting.

Distributions of the cytokine genotypes were similar between cancer patients and controls in this study and frequencies were similar to those previously published in studies of similar populations.40,41 Although we did identify an association between gastro-oesophageal cancer and the IL-1ß 511 CC genotype, this relationship lost its significance following correction for multiple comparisons. El-Omar et al. have proposed an association between IL-1ß polymorphisms and an increased risk of gastric cancer among patients with helicobacter pylori infection.47 We found no such association on sub-group analysis and other groups have similarly failed to demonstrate such an association.48,49

In summary, the possession of more than one of these genotypes (IL-6 CC, IL-10 GG and TNF{alpha} AA) resulted in a cumulative reduction in survival duration. This may relate to an increased magnitude of systemic inflammatory response or to some other unknown mechanism, but clearly there is a cytokine haplotype that is associated with adverse prognosis among these patients.


   ACKNOWLEDGMENTS
 
We thank most sincerely Mr Simon Paterson-Brown and Mr Andrew de Beaux, Consultant Surgeons, Edinburgh Royal Infirmary, for help in the recruitment of patients and provision of assistance with the clinical aspects of this work.

Received for publication March 16, 2006. Accepted for publication June 13, 2006.


   REFERENCES
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 

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D. C. Deans, B. H. Tan, J. A Ross, M. Rose-Zerilli, S. J Wigmore, W M. Howell, R. F Grimble, and K. C. Fearon
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