original papers - Advances in Clinical and Experimental Medicine

original papers - Advances in Clinical and Experimental Medicine

orIginal papers
Adv Clin Exp Med 2013, 22, 5, 683–691
ISSN 1899–5276
© Copyright by Wroclaw Medical University
Dorota DiakowskaA–D, Andrzej LewandowskiA, E, F, Krystyna Markocka-MączkaE, F,
Krzysztof GrabowskiE, F
Concentration of Serum Interleukin-27 Increase
in Patients with Lymph Node Metastatic
Gastroesophageal Cancer
Zwiększenie stężenia interleukiny 27
w surowicy pacjentów chorych na raka przełyku
i raka połączenia przełykowo-żołądkowego
z przerzutami do węzłów chłonnych
Department of Gastrointestinal and General Surgery, Wroclaw Medical University, Poland
A – research concept and design; B – collection and/or assembly of data; C – data analysis and interpretation;
D – writing the article; E – critical revision of the article; F – final approval of article; G – other
Abstract
Background. Cytokines, one of the key mediators of immune response, play an important role in cancer development.
Objectives. The aim of this study was to evaluate the interleukin-27 (IL-27) concentration in the serum of patients
with gastroesophageal cancer (GEC) and in patients with non-cancerous benign diseases of the upper digestive
tract (NCD). We investigated the relationship between the serum IL-27 level and clinicopathological factors, and
also the diagnostic utility of IL-27 as a marker of GEC presence. Additionally, we evaluated the concentrations of
serum IL-27 in patients with esophageal squamous cell carcinoma before and after surgical tumor resection.
Material and Methods. Serum samples from 84 GEC patients, 39 NCD patients and 33 healthy subjects were
assayed. The levels of IL-27, IL-6, IL-12 and IFN-γ were determined using ELISA kits.
Results. The serum levels of IL-27 were significantly higher in the GEC patients than in the healthy control
(p < 0.0001) and in NCD patients (p = 0.006). The concentrations of serum IL-27 were related to lymph node status
(p = 0.044). ROC analysis showed a significant relationship between a high level of serum IL-27 and GEC presence
(AUC = 0.766, p < 0.001). The concentrations of serum IL-27 were significantly higher in patients with esophageal
squamous cell carcinoma 3 months after esophagectomy than before the operation (p = 0.003).
Conclusions. Our results demonstrated that a high serum IL-27 level is associated with cancer presence and lymph
node metastases in GEC. Significantly higher levels of IL-27 in patients with esophageal squamous cell carcinoma
after tumor resection may imply that host immune cells are one of the important sources of circulating IL-27 (Adv
Clin Exp Med 2013, 22, 5, 683–691).
Key words: squamous cell carcinoma of esophagus, cardiae cancer, interleukin-27, tumor markers.
Streszczenie
Wprowadzenie. Cytokiny, jedne z kluczowych mediatorów odpowiedzi immunologicznej, odgrywają istotną rolę
w rozwoju nowotworu.
Cel pracy. Oznaczenie stężenia interleukiny 27 w surowicy pacjentów chorych na raka przełyku i raka wpustu
(GEC) oraz u pacjentów z łagodnymi, nienowotworowymi schorzeniami górnego odcinka przewodu pokarmowego (NCD). Autorzy poszukiwali związku między stężeniem IL-27 w surowicy a parametrami klinicznymi i patologicznymi u chorych na GEC. Przeprowadzono analizę przydatności diagnostycznej oznaczania stężenia IL-27
w surowicy jako markera obecności GEC. Dodatkowo porównano stężenia IL-27 w surowicy pacjentów chorych
na płaskonabłonkowego raka przełyku (ESCC) przed operacją usunięcia przełyku i 3 miesiące po operacji.
Materiał i metody. Zbadano surowice uzyskane od 84 pacjentów chorych na GEC, 39 pacjentów z NCD i 33 zdro-
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D. Diakowska et al.
wych osób. Stężenia IL-27 oraz innych cytokin, jak interleukiny 6 (IL-6), interleukiny 12 (IL-12) i interferonu γ
(IFN-γ) zostały zmierzone za pomocą testów ELISA. Diagnostyczna skuteczność IL-27 jako markera obecności raka
została oznaczona za pomocą analizy krzywej ROC.
Wyniki. Stężenie IL-27 w surowicy chorych z GEC było istotnie większe niż w grupie kontrolnej (p < 0,0001) oraz
u pacjentów z NCD (p = 0,006). Wykazano istotny związek stężenia IL-27 z przerzutami do węzłów chłonnych
(p = 0,044). Analiza ROC wykazała istnienie silnego powiązania między stężeniem IL-27 w surowicy a obecnością
raka u pacjentów chorych na GEC (AUC = 0,766, p < 0,001). Stężenia IL-27 były skorelowane pozytywnie ze stężeniami IFN-γ (rho = 0,34, p = 0,013) i stężeniami IL-12 (rho = 0,28, p = 0,048) w surowicy chorych na GEC. Stężenia
IL-27 były istotnie większe u pacjentów chorych na płaskonabłonkowego raka przełyku (ESCC) 3 miesiące po ezofagektomii niż przed operacją (p = 0,003).
Wnioski. Duże stężenia IL-27 w surowicy pacjentów chorych na GEC wiążą się z obecnością nowotworu i przerzutami do węzłów chłonnych. Istotnie większe stężenie IL-27 u pacjentów z ESCC po operacji wskazuje, że głównym
źródłem IL-27 mogą być komórki systemu obronnego organizmu (Adv Clin Exp Med 2013, 22, 5, 683–691).
Słowa kluczowe: płaskonabłonkowy rak przełyku, rak połączenia przełykowo-żołądkowego, interleukina 27, markery nowotworowe.
Esophageal and gastroesophageal junction cancers are significant causes of cancer-related death
worldwide [1, 2]. The poor outcome observed in
the majority of gastroesophageal cancer (GEC) patients results from the advancement of disease development at clinical diagnosis [1–3].
Squamous cell carcinoma of the esophagus
and adenocarcinoma of the distal esophagus and
gastroesophageal junction are related to chronic inflammation of differing etiologies [2]. Tumor
cells cause systemic changes in the immunological profile and in cytokines and growth factors secretion [2, 4]. For that reason, the identification
of biological markers in serum is an ideal method for early determination and monitoring of patients with GEC.
Interleukin-27 (IL-27) is a novel member of
the IL-6/IL-12 family. It is a heterodimeric cytokine composed of the EBV-transformed gene 3
subunit and p28 subunit [5]. IL-27 is predominantly synthesized by activated antigen-presenting cells including monocytes, endothelial cells
and dendritic cells [5, 6]. Although IL-27 can have
proinflammatory effects, most studies point at the
important role of IL-27 as an immunosuppressor [7, 8]. This cytokine is associated with early
phase differentiation of Th1 lymphocytes, inhibition of Th2 humoral immune response and stimulation of NK cells [5, 7–9]. IL-27 also synergizes
strongly with IL-12 in IFN-γ production via T and
NK cells (10). It has been reported that IL-27, like
IL-12, has potent anti-tumor and anti-metastatic
activity through its anti-angiogenic properties [5,
7–10].
Receptors for IL-27 (IL-27R) were composed
of a unique WSX-1⁄TCCR and common gp130
subunits [11]. The gp130 is also a receptor subunit
for IL-6 family cytokines and through this, IL-27
belongs to the IL-6 family [11].
There are some studies on the levels of cytokines such as IL-6 and IL-12 in gastric and esophageal cancer [12–14]. However, the role of soluble
IL-27 as a possible diagnostic marker in GEC remains unclear. In the present study we examined
the concentrations of IL-27, IL-12, IFN-γ and IL-6
in the serum of GEC patients and in patients with
non-cancerous benign diseases of the upper digestive tract (NCD). This study was designed to investigate the association of serum IL-27 concentration with clinicopathological parameters in GEC
patients. We also aimed for a possible diagnostic
utility of serum IL-27 level in the evaluation of gastroesophageal cancer.
Additionally, we evaluated levels of circulating IL-27 in patients with esophageal squamous
cell carcinoma before and after surgical tumor
resection.
Material and Methods
The authors examined 84 patients with esophageal squamous cell carcinoma (n = 59) and adenocarcinoma of the lower part of the esophagus
or gastric cardia (n = 25), who were treated in the
Department of Gastrointestinal and General Surgery from 2006 to 2011. There were 18 females and
66 males (median age: 58 years, 95% CI: 43–62) in
the study group.
UICC TNM staging system was applied [15]
for clinical and pathological staging. We examined
two patients with clinical stage I, 14 with stage II,
28 with stage III and 40 with stage IV.
Sera from 33 blood donors were used as a control
group. The group consisted of 9 females and 24 males,
median age 54 years (95% CI: 38–63). Additionally,
we analyzed 39 patients hospitalized due to planned
treatment for benign esophageal diseases (hiatal hernia, n = 14, cardiospasmus n = 12, burnt esophagus
n = 9, non-cancerous stenosis n = 4). There were
24 females and 15 males, median age 56 years (95%
CI: 49–59) in this group. In all the studied groups,
none of the examined indices correlated with age.
In the 21 patients with stage I, II and III of
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Interleukin-27 in Esophageal Cancer
esophageal squamous cell carcinoma, the serum
concentrations of IL-27 were measured twice: before the esophagectomy and 3 months after surgical resection.
The study protocol was approved by the Medical Ethics Committee of Wroclaw Medical University, Wrocław, Poland.
Blood samples were collected from the GEC
patients and NCD patients preceding any treatment. Blood from the peripheral vein was collected in sterile tubes, clotted (15 min, RT) and centrifuged (10 min, 900 × g). The obtained sera were
stored at –45ºC until assayed.
Concentrations of serum IL-27 were measured
using the ELISA test (BioLegend, San Diego, USA).
The sensitivity of the assay was 11 pg/mL, the intra-assay coefficient of variation was 4.0–5.9% and
inter-assay coefficient of variation was 4.9–5.4%.
All samples were run in duplicates.
Levels of IL-12 were assayed using an immunoenzymatic test (Bender MedSystems, Vienna,
Austria) accordingly to the manufacturer’s instructions. IL-6 and IFN-γ were determined with PeliKine Compact human cytokine ELISA kits (Sanquin, Amsterdam, Holland).
Distribution of data was analyzed using the
Shapiro-Wilk normality test. Concentrations of
all cytokines were presented as a median and 95%
CI (coefficient interval). The Mann-Whitney and
Kruskal-Wallis tests were used for the group comparisons. The Spearman’s rank correlation test was
used for the correlation analysis. Differences between paired variables were analyzed using a Wilcoxon test. Values of p < 0.05 were considered as
statistically significant. A power analysis of applied statistical tests demonstrated that all values
of power of the test were higher than 0.80.
The diagnostic utility of IL-27 was determined
by means of a Receiver Operating Characteristics
(ROC) curve analysis. The overall performance
of IL-27 was expressed in terms of the area under
the ROC curve (AUC) with 95% CI and p statistics
for the differences between calculated AUC and
AUC = 0.5. A cut-off value, sensitivity and specificity, Youden’s index (sensitivity (%) + specificity (%) – 100%), positive and negative likelihood
ratios were calculated.
The statistical analyses were performed using
STATISTICA 10.0 software (StatSoft Inc., Tulsa,
USA).
Results
The concentration of serum IL-27 was significantly higher in GEC patients in comparison with
healthy individuals: 216.3 pg/mL (132.7–379.5) vs.
37.5 pg/mL (36.9–101.8) (p < 0.0001), respectively (Table 1). Also serum concentration of IL-27 in
the NCD group increased significantly (89.0 pg/mL
(80.2–214.5)) in comparison with the control group
(p = 0.047). A comparison of IL-27 levels in GEC
patients with NCD patients showed significant differences (p = 0.006) between them (Fig. 1).
The concentrations of serum IL-6, IL-12 and
IFN-γ were significantly higher in GEC patients in
comparison with controls (Table 1).
The associations between circulating IL-27
levels and clinicopathological variables are shown
in Table 2. We observed a statistically significant
relationship between serum IL-27 level and lymph
node status (p = 0.044). However, the concentrations of serum IL-27 did not correlate with the
stage of disease progression, primary tumor extension and distant metastases.
The difference in the concentrations of serum
IL-27 between patients with stage I+II GEC and
control was statistically significant (p = 0.007).
These results suggest that serum IL-27 may be
a prospective diagnostic marker of cancer presence. ROC analysis revealed that serum IL-27 was
a good indicator of the presence of GEC (Fig. 2).
For purposes of comparison, the performances of
serum IL-6, IL-12 and IFN-γ as the other potential
markers of cancer presence were evaluated. The
overall performance of IL-27 was better than for
IL-12 and IFN-γ but slightly worse than for IL-6
(Table 3).
We evaluated the usefulness of serum IL-27
level as a marker of lymph node status (according to the clinical evaluation of lymph node metastases). The best cut-off value calculated from
the ROC analysis was 450 pg/mL for determination of lymph node involvement, AUC amount to
0.641(0.498–0.784), sensitivity was 34% and specificity was 86%.
A significant positive correlation was found between serum IL-27 and serum IFN-γ (rho = 0.34,
p = 0.013) in GEC patients. There was also a significant association between serum IL-12 and serum
IL-27 (rho = 0.28, p = 0.048).
The resection of tumors in patients with esophageal squamous cell carcinoma resulted in an increase of serum IL-27 concentration in 86% (18/21)
of patients. IL-27 level increased from 120.0 pg/mL
(95% CI: 72.1–324.7) before esophagectomy to
140.0 pg/mL (95% CI: 50.6–514.2) three months
after surgical tumor resection (p = 0.003) (Fig. 3).
Discussion
The results of the present study show that the
concentrations of serum IL-27 were significantly
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D. Diakowska et al.
Fig. 1. Serum levels of IL-27 in
GEC patients in comparison with
NCD patients and healthy subjects;
(* – statistically significant)
1000
900
GEC vs. control p < 0.0001*
NCD vs. control p = 0.047*
GEC vs. NCD p = 0.006*
800
Ryc. 1. Stężenie IL-27 w surowicy
pacjentów z GEC w porównaniu
z pacjentami chorymi na NCD
i osobami zdrowymi;
(* – istotne statystycznie)
IL-27 (pg/mL)
700
600
500
400
300
200
median
25–75%
individual data
dane indywidualne
100
0
GEC
NCD
control
Table 1. Serum concentration of IL-6, IL-12 and IFN-γ in GEC patients, NCD patients and healthy subjects (C)
Tabela 1. Stężenie IL-6, IL-12 and IFN-γ w surowicy pacjentów chorych na GEC, pacjentów chorych na NCD i osób zdrowych (C)
IL-6 (pg/mL)
GEC
(n = 84)
NCD
(n = 39)
C
(n = 33)
IL-12 (pg/mL)
IFN-γ (pg/mL)
median
(95% CI)
p-value
(wartość p)
median
(95% CI)
p-value
(wartość p)
median
(95% CI)
p-value
(wartość p)
1.3(1.6–5.8)
GEC vs. C,
p < 0.001*
GEC vs. NCD,
p = 0.003*
NCD vs. C,
p = 0.002*
2.7(2.8–5.7)
GEC vs. C,
p = 0.024*
GEC vs. NCD,
p = 0.815
NCD vs. C,
p = 0.078
8.8(8.6–9.8)
GEC vs. C,
p = 0.011*
GEC vs. NCD,
p = 0.986
NCD vs. C,
p = 0.083
0.7(0.0–6.9)
0.4(0.3–0.8)
2.9(2.2–5.6)
2.3(1.2–3.1)
8.3(8.0–10.9)
7.6(6.7–8.5)
* – statistically significant.
* – istotne statystycznie.
Fig. 2. ROC curves for serum IL-27 and IL-6,
IL-12, IFN-γ as possible indicators of GEC
presence
1,0
Ryc. 2. Krzywe ROC dla IL-27, IL-6, IL-12
i IFN-γ w surowicy jako prawdopodobnych
markerów obecności GEC
sensitivity
0,8
0,6
0,4
0,2
IL-27
IL-6
IL-12
IFN-γ
0,0
0,0
0,2
0,4
0,6
specifity
0,8
1,0
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Interleukin-27 in Esophageal Cancer
Table 2. Relationship between serum IL-27 concentrations and characteristic or disease-related variables in GEC patients
Tabela 2. Związek między stężeniem IL-27 w surowicy a parametrami klinicznymi i patologicznymi pacjentów z GEC
Variables (Zmienne)
IL-27 (pg/mL)
median (95% CI)
p-value (wartość p)
Gender (Płeć)
females (kobiety) (n = 18)
males (mężczyźni) (n = 66)
280.0 (207.3–509.1)
180.0 (110.4–375.8)
0.494
Age – years (Wiek – lata)
< 60 (n = 48)
> 60 (n = 36)
253.8 (64.9–322.9)
341.3 (270.5–527.3)
0.361
Histology (Histologicznie)
SCC (n = 59)
ADC (n = 25)
230.0 (172.2–351.0)
325.5 (245.6–502.9)
0.116
TNM Stage (Stopień TNM)
I+II (n = 16)
III (n = 28)
IV (n = 40)
220.0 (144.8–361.1)
255.0 (107.9–496.3)
206.3 (135.9–459.4)
0.535
Tumor (Guz pierwotny)
T1+T2 (n = 18)
T3 (n = 20)
T4 (n = 46)
260.0 (137.1–490.0)
250.0 (150.9–398.8)
253.8 (117.1–422.1)
0.342
Lymph node metastases (Przerzuty do węzłów chłonnych)
N0 (n = 28)
N1 (n = 56)
130.0 (100.6–341.2)
270.3 (259.0–435.0)
0.044*
Distant metastases (Przerzuty odległe)
M0 (n = 52)
M1 (n = 32)
190.0 (189.0–369.0)
256.3 (239.5–481.6)
0.144
SCC – squamous cell carcinoma, ADC – adenocarcinoma, * – statistically significant.
SCC – rak płaskonabłonkowy, ADC – gruczolakorak, * – istotny statystycznie.
IL-27 (pg/mL)
higher in GEC patients in comparison with the
healthy control group. This result suggests that
IL-27 overexpression may be attributed to GEC
development. The levels of serum IL-27 in GEC
were significantly higher than those in NCD patients. This may indicate that IL-27 takes part in
the inflammatory processes of the esophagus and
gastric cardiae and in gastroesophageal cancer proinflammatory reactions.
IL-27 plays multiple roles in modulating immune response. This cytokine promotes inflammation by inducing the proliferation of naive
CD4+ T cells (STAT1 and STAT3 signaling pathways) and early Th1 differentiation [6–9, 11]. On
Fig. 3. Serum IL-27 concentrations in 21 patients with esophageal squamous cell carcinoma before and 3 months after surgical
resection of tumor
before
przed
after
po
Ryc. 3. Stężenia IL-27 w surowicy 21 pacjentów chorych na
płaskonabłonkowego raka przełyku przed operacją i 3 miesiące
po resekcji guza
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D. Diakowska et al.
Table 3. Evaluation of serum IL-27 as a possible indicator of cancer presence in comparison with IL-6, IL-12 and IFN-γ
Tabela 3. IL-27 w surowicy jako prawdopodobny wskaźnik obecności raka w porównaniu z IL-6, IL-12 and IFN-γ
AUC
(95% CI)
Cut-off (Górna odcięta):
Sensitivity (Czułość) (%)
Specificity (Specyficzność)(%)
Youden’s index (Indeks Youdena) (%)
LR+
LR–
IL-27
IL-6
IL-12
IFN-γ
0.766
(0.672–0.859)
> 260 U
47.2
96.2
43.4
12.3
0.53
0.849
(0.751–0.948)
> 1.22 U
51.9
94.4
46.3
9.35
0.48
0.685
(0.517–0.853)
> 3.7 U
20.7
93.8
14.5
3.31
0.79
0.759
(0.606–0.912)
> 8.7 U
52.8
88.9
41.7
4.75
0.47
AUC – area under ROC curve, LR+ and LR– – positive and negative likelihood ratios.
AUC – pole powierzchni pod krzywą ROC, LR+ i LR– – pozytywny i negatywny współczynnik prawdopodobieństwa.
the other hand, IL-27 inhibits inflammation responses by suppressing Th2 and Th17 differentiation [16]. Analyzing the IL-27 level in NCD patients, we demonstrated a statistically significant
elevation of this cytokine in comparison with control subjects. The increased concentration of IL-27
observed in NCD patients may be connected with
the up-regulation of Th1 and down-regulation of
Th2 cell activity.
The results of the present study show the highest
level of serum IL-27 in GEC patients. Through active participation in inflammatory processes, IL-27
may directly or/and indirectly influence the initiation and development of gastroesophageal cancer.
The possible role of IL-27 in GEC may be explained
by the participation of this cytokine in the initiation of Th cell response. IL-27 generates a Th1 immune response, which influences the stimulation of
the secretion of proinflammatory cytokines such as
TNF-α, IL-1, IL-6 and IL-8 [8, 17]. IL-27 activates
the production of Th1-type cytokines such as IL-12,
IL-18 and IFN-γ, and inhibits secretion of Th2-type
cytokines such as IL-4 and IL-10 [16, 18]. It has been
shown that IL-27 plays an important role in the suppression of Th2 response and induction of Th1 response [5, 8, 10, 18]. Through stimulation of proinflammatory cytokines and induction of CD8+ T and
NK cell activity, IL-27 has prospective anti-tumor
and anti-angiogenic effects [7, 8, 16]. Selective usage of these anticancer mechanisms depends on the
properties of tumor cells [7, 16].
Many studies analyze biological prognostic
markers, which in connection with clinicopathological parameters modify the staging systems for optimal therapy for patients. Our studies are the first ones
which analyze serum IL-27 concentration in patients
with esophageal and cardiae cancer to determine the
clinicopathological and prognostic significance of this
cytokine. We demonstrated a significant relationship
between a high level of serum IL-27 and lymph node
metastases (LNM) in GEC patients.
Early lymphatic invasion is characteristic for
esophageal squamous cell carcinoma and its detection is significant in the course of esophageal cancer [19]. However, precise clinical lymph
node staging is very difficult. With respect to this,
a non-invasive biomarker predicting LNM may be
a helpful prognostic factor in cardiae and especially in esophageal cancers. However, our analysis of
serum IL-27 level as a marker of lymph node status did not show satisfactory results.
Recently, the dependence of LNM to T tumor
stage has been reported [20]. It has been observed
that tumor invasion is a predictor of lymph node
metastases. But in our study, IL-27 level was not
significantly different in relation to the depth of
primary tumor invasion and cancer stage. With
respect to these results, we did not examine IL-27
concentration in GEC according to their T-LNM
status.
In vitro study on multiple myeloma, colon carcinoma tumor model and neuroblastoma tumor
cells demonstrated that IL-27 could exert a potent anti-tumor effect [8, 21, 22]. Studies of Hu
et al. [23] have demonstrated that transfection of
IL-27 gene into a murine hepatocellular carcinoma
cell line prevents tumor growth. In contrast, in vitro studies of Lo et al. [24] have shown that IL-27
has a slight influence on both innate and adaptive lymphocytes. The exact mechanisms of these
opposite effects of IL-27 are not clear and need
further exploration. It is possible that IL-27, like
IL-12, mediates activation of anti-tumor immunity by inducing neoplastic cells to produce anti-angiogenic factors [25].
However, on the basis of the above-mentioned
observations and our data, we assume that IL-27
may possibly induce the differences in local proinflammatory cytokine levels in a tumor microenvironment and simultaneously may activate host
immune cells in the surrounding tissues against
tumor growth and metastases.
Interleukin-27 in Esophageal Cancer
The authors observed that the level of serum
IL-27 increased significantly in stage I+II of cancer in comparison with the control, and remained
high in stages III and IV of GEC. This may indicate that IL-27 up-regulation is associated with
early neoplastic transformation and disease progression, probably by activation of host immune
response against cancer.
To the best of our knowledge, we are the first
who have evaluated the diagnostic parameters,
such as sensitivity, specificity, positive and negative predictive value and ROC curve, for IL-27 in
GEC patients. We showed that the IL-27 area under the ROC curve was higher than AUC for IL-12
and IFN-γ, but lower than AUC for IL-6. Also,
prognostic sensitivity, specificity and Youden’s index for IL-27 were lower than for IL-6. We compared our results for IL-27 with data for classic
tumor markers in the serum of esophageal and
gastric cancer patients [26, 27]. We revealed in our
analysis that AUC for cell cancer antigen (SCCAg) (AUC = 0.811, p < 0.001) was higher than
AUC for IL-27. Simultaneously, AUC for IL-27
was higher than AUC for carcinoembryonic antigen (CEA) (AUC = 0.673, p < 0.001) and carbohydrate antigen (CA19-9) (AUC = 0.762, p < 0.001).
These observations suggest that high serum IL-27
level may participate in GEC presence. Further investigations are necessary to assay the role of IL-27
as a prognostic parameter in upper gastrointestinal tract cancers.
This is the first report about differences between concentrations of IL-27 in patients with
esophageal squamous cell carcinoma before and
after surgical tumor resection. In the majority of
patients, the level of IL-27 was higher after tumor
resection than before esophagectomy. This observation may be consistent with our hypothesis that
host immune cells can significantly influence serum IL-27 concentration and that the primary tumor is not the only important source of circulating
IL-27 in esophageal squamous cell carcinoma. It is
also possible that non-detected lymph node metastases and micrometastases can induce synthesis of
IL-27 in patients after esophagectomy.
Further studies are necessary for better clarification of the main source of high IL-27 level in
gastric and esophageal cancers. In our future research, we would like to compare IL-27 expression
in tumor tissue with the tissue surrounding the tumor in GEC patients.
Chronic inflammation, in which many pro-inflammatory cytokines take part, may be a risk factor for cancer development [2, 3, 16]. We marked
the IL-6 and IL-12 serum levels as cytokines which
belong to the same family as IL-27, and IFN-γ serum level as well. Our data showed that serum
689
IL-6 levels were significantly higher in GEC patients than in healthy subjects. We reported similar accuracy in IL-6 detection to those described
for esophageal and gastric cancer by other authors [26], where serum concentrations of IL-6
significantly increased. IL-6 plays an important
role in host defense mechanisms. This cytokine is
able to stimulate the anti-tumor activity of macrophages and to induce the expression of vascular
endothelial growth factor [28]. However, overexpression of IL-6 mRNA and the protective role of
IL-6 from apoptosis have been observed in esophageal carcinoma cell lines [18]. Recently Guzzo et
al. [29] have demonstrated that IL-27 induces IL-6
expression in human monocytic cells. Our correlation studies did not find a significant correlation
between serum IL-6 and IL-27.
The anti-tumor activity of IL-12 is highly dependent on primary IFN-γ produced by T and NK
cells in response to IL-12 [7, 9, 10, 25]. IFN-γ plays
an important role in enhancing antigen specific and nonspecific immune responses which support tumor rejection [25]. In the present study, we
observed significantly higher levels of serum IL-12
and IFN-γ in GEC patients in comparison with
the control group. These findings are in line with
our previous studies, where we demonstrated that
the serum level of IL-12 was significantly higher in
squamous cell carcinoma of the esophagus than in
healthy controls [14]. It may be possible that IL-12
and IFN-γ participate in GEC anti-tumor immunity by inhibition of tumor cells in proangiogenic
factor production.
Because IL-27 also induces synergistic production of IFN-γ with IL-12 [10, 25] it was expected
that the anti-tumor activity of IL-27 might be maintained. In experimental studies, Shimizu et al. [7]
showed the strong inhibition of neovascularization
by IL-27 in chicken embryos. They said that IL-27
had a potent anti-angiogenic activity, and this effect was IFN-γ independent. These results suggest
that IFN-γ and IL-12 do not require IL-27 for the
stimulation of anti-metastatic and anti-angiogenic
activities. Our opposite results revealed that serum
IL-27 levels correlate significantly with serum levels of IFN-γ and with serum IL-12 in GEC patients.
The studies of Xu et al. [16] have also indicated that
IL-27 exerts an anti-tumor effect via CD8+ T cells
and IFN-γ production in highly immunogenic tumors expressing MHC class I. Therefore, we suggest that IL-27 enhances anti-tumor and anti-angiogenic activity through IFN-γ production in GEC
patients.
In conclusion, our results demonstrated the
stimulation of IL-6, IL-12, IL-27 and IFN-γ production in GEC patients. High IL-27 level was positively associated with the presence of lymph node
690
D. Diakowska et al.
metastases in GEC patients. ROC analysis showed
a significant relationship between a high level of serum IL-27 and GEC presence. However, possible application of IL-27 as a diagnostic marker of cancer
presence warrants further investigation. Significantly higher levels of IL-27 in patients with esophageal
squamous cell carcinoma after tumor resection may
imply that host immune cells might significantly influence serum IL-27 concentration. A significant
correlation between serum IL-27 and serum IFN-γ
suggests a possible relationship between these interleukins activation and production in GEC.
Acknowledgements. The authors would like to thank dr Elżbieta Klausa from the Regional Center of
Blood Donation and Therapeutics in Wrocław, Poland for supplying the serum of healthy individuals.
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Address for correspondence:
Dorota Diakowska
Department of Gastrointestinal and General Surgery
Wroclaw Medical University
Sklodowska-Curie 66
50-369 Wroclaw
Poland
Fax: +48 71 733 26 09
E-mail: [email protected]
Conflict of interest: None declared
Received: 9.10.2012
Revised: 1.02.2013
Accepted: 3.10.2013