Cancer Investigation, 32:197–205, 2014 ISSN: 0735-7907 print / 1532-4192 online C 2014 Informa Healthcare USA, Inc. Copyright DOI: 10.3109/07357907.2014.898156
REVIEW
Interleukin-8 and Interleukin-17 for Cancer Paul Zarogoulidis,1 Fotini Katsikogianni,2 Theodora Tsiouda,3 Antonios Sakkas,4 Nikolaos Katsikogiannis,2 and Konstantinos Zarogoulidis1
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1
Department of Pulmonary, Oncology Unit, “G. Papanikolaou” General Hospital, Aristotle University of Thessaloniki, Thessaloniki, Greece; 2 Department of Surgery (NHS), University General Hospital of Alexandroupolis, Alexandroupolis, Greece; 3 Department of Internal Medicine, “Theageneio” Anticancer Hospital, Thessaloniki, Greece; 4 Department of Pathology, “G. Papanikolaou” General Hospital, Thessaloniki, Greece been associated with advanced stage disease, identified as a prognostic factor (8, 9), and as a possible treatment target through inhibition of IL-8 production and blockage of IL-8 receptors (10, 11). Increased levels have been observed and associated with advanced stage and poor prognosis in cyst fluid, ascites, additionally over-expression of IL-8 has been observed in tumor tissue. The activity of IL-8 is mediated through the receptors IL-8RA and IL-8RB. The first is binding only to IL-8 and granulocyte chemotactic protein-2, while the second with neutrophil activating peptide, granulocyte activation peptide-2, and chemokine Gro-α,β,γ (10). The IL-8RA has been also associated with the pathogenesis of serous tumors (12). IL-8 stimulates proliferation by increasing levels of Cyclin D1 and Cyclin B1 and via activation of Phosphatidylinositide 3-kinases PI3K/Akt and Extracellular signal-regulated kinases Raf/MEK/ERK. Therefore, the mesenchymal transition occurs (13). The invasive capability of a tumor has also been found to be mediated with the increased activation of matrix metalloproteinase-2 and 9 (MMP-2,-9) that are mediated by the levels of IL-8 (14). Therefore, IL-8 levels have been associated with metastatic invasiveness and early recurrence (15). IL-8 expression has been found to be lower in normal cells (10). The MMP as proteolytic enzymes that have the ability to degrade basal membrane. The MMPs that exist in the extracellular matrix play a key role in tumor metastasis. However, although there are 25 MMPs, MMP-9 has been found over-expressed in cancer tissue (16). Further experimentation is overexpression of MMP-9 in a prostate cancer model confirmed the association of MMP-9 with tumor invasiveness (17). It has also been found in a gastric carcinoma model that IL-8 upregulates MMP-9 expression and consequently increased neoangiogenesis is observed (18). IL-17 is a pro-inflammatory cytokine group of ligands that is primarily produced from a subset of CD4+ effector cells known as Th17cells (19). IL17 has been previously associated with the pathogenesis of inflammatory diseases (20). IL-17 induces the expression of
Pro-inflammatory cytokines have been associated with chronic inflammation and inflammatory diseases. Increased levels of interleukins (ILs) have been associated with inflammatory disease exacerbation. ILs levels have been observed to be associated with advance stage cancer for several types of cancer and a poor prognostic maker for malignant disease. Moreover; increased levels of cytokines induce tumorigenesis. There are several paradigms such as the hepatocellular carcinoma induced from chronic inflammation of an underlying hepatitis. In the current review, we will focus on IL-8 and -17. These two ILs as in the case of others, induce neo-angiogenesis through activation of the vascular endothelial growth (VEGF) factor pathway. Additionally, they enhance the activity of matrix metalloproteinase-2 and –9 (MMP-2,-9) which in turn increase the metastatic activity of the underlying malignancy. Inhibition of cytokine production could be a potential treatment both for chronic inflammatory diseases and tumor modulation. Local microenvironment modulation could be applied in surgery resected patients as in the case of lung cancer in order to enhance the local immune activity. Keywords: Interleukin-8, Interleukin-17, Cancer, Immunomodulation
INTRODUCTION Interleukin-8 (IL-8) is a multifunctional pro-inflammatory cytokine produced under inflammation stimulation and is attracting and activating neutrophilic granulocytes (1). Il-8 is secreted by numerous cells such as; (a) mesothelial cells, (b) various tumor cells, (c) neutrophils, (d) endothelial cells, and (e) monocytes (2). It has been characterized as an; angiogenenic factor, motogenic factor, autocrine, and paracrine growth factor. This ability has been observed in numerous cancer types such as; (a) melanoma, (b) prostate cancer, (c) colon cancer, (d) ovarian cancer, (e) nasopharyngeal, (f) lung cancer, and (g) hepatocellular (3–7). IL-8 levels have
Correspondence to: Paul Zarogoulidis, M.D, Ph.D., Pulmonary Department-Oncology Unit, “G. Papanikolaou” General Hospital, Aristotle University of Thessaloniki, Thessaloniki, Greece, email: [email protected] Received 3 January 2014; revised 25 January 2014; accepted 17 February 2014.
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P. Zarogoulidis et al.
chemokines; CXCL1, CXCL6, CXCL8, nuclear factor-kappa B (NF-κB), growth-colony stimulating factor (G-SCF), granulocyte macrophage-colony stimulating factor (GM-CSF), adhesion molecules (ICAM-1), and IL-6 (21, 22). It has been observed that different polymorphisms are associated with different autoimmune diseases such as; rheumatoid arthritis and ulcerative colitis, therefore it has been suggested that the aberrant expression of IL-17 induces abnormal immune response (20). IL-17 has been observed to act upon different cell types (19, 23). The production of IL-17 has been also associated with helicobacter pylori infection and consequently with underlying disease progression (24). IL-17 levels have been found increased in colon (25), ovarian (26), hepatocellular (27), esophageal (28), lung cancer (29), and glioblastoma (30). IL-17 polymorphisms have been associated with gastric cancer through overproduction of IL-17, which in turn over-stimulates the immune system (31, 32). There are controversial data indicating that there is probably a cut off point in the production of IL-17, which if increased it overstimulates the local microenvironment and instead of protecting from mutagenesis it induces it (33). It has been previously observed that there are increased circulating levels of IL-17-producing Th-17 cells in patients with gastric cancer (34). In this subset of patients, colonization with helicobacter pylori does not play a role in gastric cancer development. In colorectal cancer, IL-17 expression was observed to be mediated by ras signaling, implying a new treatment with downregulation of this pathway (35). Furthermore, it has been observed that metastasis is associated with increased levels of IL-17 via activation of CXCR-2-dependent neo-angiogenesis (36). Increased levels of IL-17 have been associated with poor survival and metastasis through increased lymphangiogenesis (37, 38). Increased levels of IL-17 have been found in advance stage disease (29) and could be used as a prognostic marker (39). Increased levels of IL-17 have been associated with early recurrence (40). IL-17 and IL-10 have found elevated in malignant pleural infusion presenting remaining evidence of a mutagenesis process based on inflammation (41). Also in the study by Li et al. (29) it was observed that IL-17 induced metastasis by acting and over-expressing the IL-6 pathway (42, 43). In the current work, we will focus on the clinical data regarding these two pro-inflammatory cytokines. RESEARCH STRATEGIES We performed an electronic article search through PubMed, Google Scholar, Medscape, and Scopus databases, using combinations of the following keywords: IL-8 and cancer, IL-17 and cancer. All types of articles (randomized controlled trials, clinical observational cohort studies, review articles, case reports) were included. Selected references from identified articles were searched for further consideration, without language limitation (Table 1). IL-8 In the study by Shiau et al. (44), it was observed that human papillomavirus (HPV) infection is associated with the activ-
Table 1. Reference Selection Libraries: PubMed, Google Scholar, Medscape, and Scopus databases Key words: interleukin-8 and cancer, interleukin 17 and cancer 1rst Step: 70 studies initially identified and screened 2nd Step: 24 studies excluded by title or abstract examination 3rd Step: 23 studies were identified to be relevant from the references of the 46 previously identified studies 4th Step: 69 references were finally chosen to be included
ity of MMP-2 and -9 that are known to be associated with tumor invasiveness. Moreover; the IL-8 levels were again associated with HPV, neo-angiogenesis, and consequently tumor proliferation. This observation was performed in female never smokers with nonsmall cell lung cancer (NSCLC). In the study by Singh et al. (45), it was observed that IL-8 activity plays a crucial role in breast cancer, it is regulated by Human Epidermal Growth Factor Receptor 2 (HER-2) positive cancers through CXCR1/2 ligands. The IL-8 levels influence the breast cancer stem cell activity by enhancing or downregulating tumorigenesis. Therefore, IL-8 targeted therapy could potentially have a place for breast cancer, however; since it is also regulated by HER-2 expression then conventional therapy could also be sufficient. The activity of MMP-9 and IL-8 were blocked in the study by Park et al. (46) with luteolin 8-C-β-fucopyranoside. In specific, the following pathways were inhibited via ERK/AP-1 and ERK/NF-κB. Metastasis and tumorigenesis were blocked with LUC8C-FP. The polymorphism of IL-8–251 A/T in NSCLC was investigated in a large population study and it was observed that it is not associated with NSCLC. This observation indicates that in specific this expression of pro-inflammatory cytokine is not associated with tumorigenesis (47). However, in the study by Rafrafi et al. (48) high expression of IL-8mRNA was observed in lung cancer patients, which was further associated with tumor size and advance stage disease. In this study the IL-8–251T/T polymorphism was associated with increased tumor size and advance stage lung cancer. Also, this polymorphism was associated with NSCLC adenocarcinoma. In the study by Lattanzio et al. (49) the effect of radiotherapy and anti-angiogenic in the modulation of IL-8 activity was investigated. It is well known that IL-8 increased levels are associated with resistance to anti-angiogenic therapies, this observation was made with bevasizumab therapy. The combination of radiotherapy and antivascular endothelial growth factor (VEGF) reduce effectively the IL-8 levels and its activity. In the study by Yamada et al. (50), biopsies were taken from healthy volunteers and patients with advanced gastric cancer. It was observed that IL-8 mRNA expression level, Helicobacter pylori infection and low PG I/II ratio could be a prognostic cluster without any correlation with cagA mutation. In the study by Browne et al. (12), the levels of IL-8 levels and expression of IL-8RA and IL8-RB were evaluated as a potential prognostic markers for ovarian cancer and risk or tumorigenesis. The IL-8 levels and IL-8RA expression were observed to be potential prognostic biomarkers and were also suggested to be therapeutic targets in ovarian serous carcinoma. In the study by Albulescu et al. (51), the following Cancer Investigation
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IL-8 and IL-17 parameters were investigated in brain cancer: IL-1β, IL-2, IL4, IL-6, IL-8, IL-10, IL-12, GM-CSF, VEGF, Fibrotic growth factor-2 (FGF-2), and TNF-α. The IL-6, IL-10, TNF-α, and IL-1β were upregulated by threefold and VEGF, FGF-2, IL-8, IL-2, and GM-CSF were upregulated by twofold. Therefore, it can be said that in brain cancer strong inflammatory response along with enhanced tumorigenesis is observed. In the study by Jo et al. (52), the blood flow was correlated with IL-8 in order to identify whether it could be correlated with treatment efficiency. The concept is that there are agents targeting Bcl-2 pro-angiogenic pathway that could be used when there is elevated blood flow. In the study by Desai et al.(53), the concept of inhibiting IL-8 was again evaluated as a possible treatment strategy in lung cancer. Data presented again the close association of VEGF with IL-8 levels, moreover; the mesenchymal transition and invasiveness was again associated with IL-8 levels. This observation can be attributed to the MMP-2 and -9 enhanced activity due to the elevated levels of IL-8. The levels of IL-8 and MMP-9 expression were identified as prognostic markers for survival and tumor invasion in nasopharyngeal carcinoma (5). The increased levels of IL-8 induce resistance to anti-VEGF therapy, therefore, a dual therapy of blocking IL-8 and concominately administration of anti-VEGF administration could be potentially an efficient therapy as indicated in the study of Gyanchandani et al.(54). In the study by He et al. (55), tumorigesis was induced with hexavalent chromium Cr (VI) and neoangiogenesis was observed to be correlated with IL-8 leveles and insulin like growth factor-1 (IGF-1). The tumorigenesis of Cr (VI) was regulated by miR-143 expression, by reducing this expression tumorigenesis occurred. In the study by Hsiao et al. (56), it was observed that IL-6 and IL-8 levels are upregulated in the presence of E6 and E7 genes of HPV type 18. Moreover, it was observed that UV radiation increases the activity of E6 and E7 genes. In the study by Hsu et al. (57), it was observed that in epidermal growth factor receptor IL-8 levels and cycloxygenase (COX-2) are overexpressed and downregulated by a coral natural product. The downregulation was achieved through targeting Ca2+ signalingdependent inflammatory diseases. In the study by Cesaratto et al. (58), the APE1 over-expression was associated with chemo-resistance and onset inflammatory response in liver cancer cell lines (Table 2). IL-17 IL-17 family has been associated with numerous inflammatory diseases (20), additionally as it will be described below there, it is a crucial modulator for tumorigenesis and metastasis. In the study by Li et al. (29), it was observed that increased levels of IL-17 were associated with advanced stage disease in lung cancer. IL-7R increased expression was also associated with increased invasiveness. The Signal transducer and activator of transcription 3 (IL-6-STAT3) pathway was activated by the increased levels of IL-7 in order for NSCLC cells to increase their invasiveness. In the study by Petanidis et al. (35), an association of the k-ras status was observed with disease status in colorectal cancer. Moreover, k-ras mutation status was associated with the expression of C 2014 Informa Healthcare USA, Inc. Copyright
interferon-γ (IFN-γ ) and granulocyte macrophage-colony stimulate factor (GM-CSF). K-ras positive patients have the IL-17, IL-22, and 23 differently expressed and the increased levels induce tumorigenesis. The polymorphism G-197A in the IL-17 promoter region was associated with increased gastric cancer risk independently of Helicobacter pylori presence. Moreover, in homozygous patients (AA) at position -197 the chance of developing gastric cancer was 2.9 times higher than single (A) (59). In the study by Wu et al. (40), increased pretreatment levels of IL-17 were associated with poor prognosis of hepatocellular carcinoma and additionally; increased IL-17 levels were associated with early recurrence. In the study by Klimatsidas et al. (41), an association between IL-10, IL-17, and malignant pleural effusion was presented. In specific, it was suggested that the levels measured in the pleural fluid were a result of an inflammatory/mutagenic process. These levels were collated with malignancy type. In the study by Zhang et al. (39), it was observed that TNM stage, positive IL-17, Foxp3 expression, and negative CD4+ were the main prognostic factors. Patients survival undergoing surgery can be more efficiently assessed when combining clinical staging and IL-17/Foxp3 expression. The association of IL-17 expression and levels has been observed in liver fibrosis (60). The IL-6 has been previously also associated with activation of fibrotic mechanisms when cancer patients receive tyrosine kinase inhibitors (TKIs) (42, 43). Since IL-17 uses the IL-6 STAT pathway to enhance the tumor invasion capability it could be stated that these two IL are closely related at least from a clinical point of view. Further investigation of the interaction between these two pathways is needed. The IL-17-producing Foxp3+CD4+ T cells have been observed to play a crucial role in tumorigenesis of colorectal cancer and initiation of chronic bowel disease (61). In the study by Olsson-Akefeldt et al. (62), IL-17 effect on the tumor microenvironment was evaluated and controversial results were presented. Again it seems that the levels of IL-17A play a dual role of enhancing the immune system against tumorigenesis, however; probably after a cut off point that has not been established until now they induce tumorigenesis. In the study by Hayata et al. (63), the effect of IL-17 was again evaluated in the tumor microenvironment by administration of gene therapy. It was observed that at the sites of IL-17 inhibition the tumor proliferation was blocked through down-regulating neo-angionenesis and regulation of cytotoxic T lymphocytes. The MMP-9 activity that is a protein that enhances the cancer cell invasion and metastasis was also blocked with local control of IL-17. In the study by Jordan Wiliams et al. (64), the basic leucine zipper transcription factor (BATF) that dimerizes with the JUN proteins to direct patterns of activator protein-1 (AP-1)mediated gene expression was investigated. In specific, BATF was investigated how it interacts with the regulation of IL-17A and the results indicated that it could be used as a treatment target. IL-17 was also investigated in esophageal squamous cell carcinoma as a prognostic factor (28). It was observed that mast cells play a crucial role in tumor immunity by releasing IL-17 in muscularis propia and therefore
Colorectal patients
IL-17 G-197A polymorphism (patients) Hepatocellular
Malignant pleural fluid Lung cancer
Liver fibrosis Dendritic cells
Colon
Mice expressing human HA-tagged BATF Esophageal squamous cell carcinoma
Glioblastoma (patients) Colon cancer
Ovarian (patients)
Hepatocellular (Patients)
Breast (patients) Ovarian (patients)
Lung cancer
Breast cancer NSCLC Uveal melanoma
Petanidis et al.
Rafiei et al.
Klimatsidas et al. Zhang et al.
Du et al. Olsson-Akefeldt et al.
Hayata et al.
Jordan-Williams et al. Wang et al.
Cui et al. Straus D.
Lan et al.
Liao et al.
Chen et al. Droeser et al.
Shiau et al.
Singh et al. Bhat et al. Lattanzio et al.
Wu et al.
NSCLC patients
Cancer
Li et al.
Author
Table 2. Clinical Studies
ELISA, CM and Flow cytometry, tissue microarray design and immunocytochemistry IHC, quantification of staining results Tissue microarray construction, IHC, validation analysis of IL-17 staining Zymography, Western blot, RNA extraction and RT-PCR, ELISA Review DNA extraction, PCR, ELISA, western blot
IHC, immunofluorescence, immunoelectron, quantification of immunofluorescence, evaluation of immunohistology IHC, Glucose and L-lactate assays, Growth factors assays, Protein experiments and AKT phosphorylation assays. IHC, selection of the cut-off point for expression of IL-17
ELISA, RT-PCR, IHC TRAP and Hoechst staining, CFSE and CD14-PE labeling for proliferation study, Flow cytometry, western blot, immunocytofluorescence labeling, affymetrix genechip study, RT-PCR, Functional analysis Cytotoxic assay, flow cytometry analysis, IHC and quantitative microscopy, Measurement of cytokine and chemokine content in tumor tissues and in vitro ELISA, Flow Cytometry
ELISA IHC, Eli Vision two-step kit
Quantification of serum levels of cytokines, FACS
DNA-RNA extraction, ELISA, RT-PCR, Westen blot, IHC, MTT cell viability, SPOT GM-CSF-IFN-γ , TUNEL assay, K-ras status IL-17 genotyping, Helicobacter pylori detection
Invasive assay, Plasmid construction, western blotting, evaluation of tumor progression in vivo
Method of Evaluation
Results
IL-8 levels associated with neo-angiogenesis, MMP-2 and -9 associated with tumor invasiveness-metastasis IL-8 levels regulate breast cancer stem cell activity. IL-8–251 A/T IL-8 levels and signaling were more effectively reduced when combination of radiotherapy and bevacizumab were administered
IL-17 is a good prognostic marker for glioblastoma TNF-α and IL-17 increased levels enhance tumorigenesis in colorectal cancer IL-17 and TAM density are correlated. IL-17 levels are associated with PFS and survival IL-17 and IL-17RE increased levels are a poor prognostic factor for HCC
IL-17 increased in propia muscularis along with mast cells, macrophages and CD8+ T cells
IL-17 inhibition with gene therapy down regulates neo-angiogenesis and MMP-9 activity, therefore inhibiting tumorigenesis BTAF could be a possible treatment target
G-197A polymorphism associated with development of gastric cancer Increased pretreatment levels associated with poor prognosis and early recurrence IL-10, IL-17 levels associated with malignancy TNM staging, IL-17 and Foxp3 expression, CD4+negative expression associated with disease prognosis IL-17 levels associated with liver fibrosis Modulation of microenvironment
IL-17 levels associated with advance stage, IL-17R increased expression associated with increased invasiveness. K-ras positive mutation associated with increased tumorigenesis
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(45) (47) (49)
(44)
(37) (65)
(27)
(26)
(30) (25)
(14)
(64)
(63)
(60) (62)
(41) (39)
(40)
(59)
(35)
(29)
Ref
P. Zarogoulidis et al.
Cancer Investigation
C 2014 Informa Healthcare USA, Inc. Copyright
Gastric cancer
Ovarian cancer
Breast cancer
Brain cancer (patients)
Head and Neck squamous cell carcinoma (patients) Lung cancer
Yamada et al.
Browne et al.
Park et al.
Albulescu et al.
Jo et al.
Lung cancer
Basal cancer
Lung cancer
Liver cancer cell lines
Gyanchandani et al.
He et al.
Hsiao et al.
Hsu et al.
Cesaratto et al.
MTT and cell growth assays, MTS assay, Nile red staining, western blot, immunofoulorescence assay and confocal analysis, RT-PCR, soluble cytokines determination, transient transfection experiments
ELISA, RT-PCR, western blotting, luciferase assay
ELISA, antibody array, transwell migration, cell proliferation, western blot, capillary-tube formation, IHC and immunofluorescence RT-qPCR, colony isolation, tube formation assay, IHC, miRNA transfection, chorioallantoic membrane assay, miRNA luciferase reporter constructs and luciferase activity assay Editorial
Cell adhesion assay, confocal immunofluorescent staining, western blotting and IP, motility and invasion assay, gelatin zymography, anoikis assay, cell transfection, soft agar colony formation assay Serum IL-8 and MMP-9 assay
CT perfusion, serum samples and ELISA
MTS assay, wound healing assay, matrigel invasion assay, gelatin zymography, RT-PCR, ELISA, western blotting xMAP analysis and ELISA,
IHC, assessment of expression
Determination of mRNA expression, genotyping of cytokine gene polymorphisms RT-PCR, genotyping, antibody test
UV increase the activity of E6 and E7 genes in HPV type 18 virus and IL-6 and IL-8 levels are increased in the presence of E6 and E7 Coral natural product a potential treatment for increased levels of COX-2 and IL-8 APE1 responsible for chemo-resistance
miR-143 regulates tumorigenesis from Cr (VI) and IL-8 levels are associated with neoangiogenesis
IL-8 increased levels and MMP-9 expression are considered as prognostic markers IL-8 and anti-VEGF co-administration more efficient than one agent alone
Helicobacter Pylori, mRNA expression and low PG I/II ratio are associated with increased rick of gastric cancer IL-8 and IL-8RA potential prognostic markers and treatments LUC8C-FP a possible treatment for blocking metastasis and tumorigenesis IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, GM-CSF, VEGF, FGF-2 and TNF-α. The IL-6, IL-10, TNF-α, IL-1β were up-regulated by threefold and VEGF, FGF-2, IL-8, IL-2, and GM-CSF were upregulated by twofold Elevated blood flow associated possibly with effective Bcl-2 targeted treatment IL-8 possible targeted treatment which down-regulates the VEGF pathway
IL-8–251T/T association with NSCLC
(58)
(57)
(56)
(55)
(54)
(5)
(53)
(52)
(51)
(46)
(12)
(50)
(48)
NSCLC; nonsmall cell lung cancer, IL-17; interleukin-17, IL-17R; interleukin 17 receptor, ELISA; Enzyme-Linked Immunosorbent Assay, RT-PCR; Reverse transcription polymerase chain reaction, IHC; immunohistochemistry, MTT; tetrazolium dye, GM-CSF; granulocyte macrophage-colony stimulating factor, IFN-γ ; interferon-γ , TUNEL; Terminal deoxynucleotidyl transferase dUTP nick end labeling, SPOT; enzyme-linked immunosorbent spot, FACS; Fluorescence-activated cell sorting, CFSE; Carboxyfluorescein succinimidyl ester (CFSE) is a fluorescent cell staining dye, MMP-9; matrix metalloproteinase, TNF-α; tumor necrosis factor-α, TAM; tumor associated macrophages, PFS; progression free survival, HCC; hepatocellular carcinoma, CM; solid-phase cytometry, MTS; MTT, CT; computed tomography, xMAP; multiplex assay, VEGF; vascular endothelial growth factor, UV; ultra violet radiation.
Nasopharyngeal cancer (Patients) Head and neck cancer
Cheng et al.
Desai et al.
Lung cancer (patients)
Rafrafi et al.
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IL-8 and IL-17
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P. Zarogoulidis et al.
could serve as a prognostic factor for squamous cell carcinoma. Activated macrophages and increased levels of density effector CD8+ T cells were also observed in muscularis propia along with increased levels of IL-17. In the study by Cui et al. (30), the expression of IL-17 was evaluated in glioblastoma cells and was observed to be a valuable prognostic marker. In specific, higher levels of IL-17 were correlated with progression free survival (PFS), and death rate. The IL-17 and PFS were also observed to be independent factors affecting the overall survival. Therefore, it can probably be speculated that in the case of glioblastoma the IL-17 is modulating the tumor microenvironment as a defense mechanism. The same observation was made for ovarian cancer, increased levels of IL-17 were associated increased PFS and survival (26). Furthermore, it was observed that the density of tumor-associated macrophages (TAMs) was associated with the levels of IL-17. Moreover, increased levels of IL-17 have been associated with chemo-sensitivity to platinum-based chemotherapy (65). The contribution of tumor necrosis factor-a and IL-17 in colon cancer tumorigenesis was investigated, and it was observed that these pro-inflammatory cytokines contribute to cancer proliferation through stimulation of glycolysis and growth factor production (25). This study by Straus D. indicates again that IL-17 blockage could be a future treatment proposal. In the study by Liao et al. (27) the IL-17 and IL-17RE were found to be poor prognostic factors for hepatocellular carcinoma. In this type of cancer the chronic nature of the inflammation and continuous activity of IL-17 producing CD4+ T cells could be the substrate of tumorigenesis. Targeted therapy in a form of pulse adjuvant treatment could be helpful, activating the defense mechanisms but not over-expressing them. Moreover; increased IL-17 levels were associated with poor survival in breast cancer (66). Increased levels of IL-17 were also associated with HER-2, advanced T stage and presented shorter disease free survival when combined (Table 2). DISCUSSION Cytokines have been observed increased in several chronic inflammatory diseases, and increased levels have been associated with disease exacerbation. IL-8 has been extensively investigated in lung cancer, while IL-17 has been investigated in a variety of cancer types. There is the case of crohn‘s disease were treatment with monoclonal antibodies inhibiting the inflammation cascade and cytokine production have provided an efficient disease control for many patients (42, 43). Inflammatory cytokines have been observed increased in advanced stage disease for many types of cancer and increased levels are a poor prognostic marker. The IL-17 family consists of numerous subdivisions such as; A, B, C, D, E and F. The IL-17A blocking has been tested for autoimmune disorders and could be a future treatment methodology. Regarding B, C, D and E their function still remain to be elucidated. IL-17F plays a crucial role in host defense from bacteria (20). The modulation of the tumor microenvironment is possible through inhibition of IL-17 activity. The inhibition of IL-8
Figure 1. IL; interleukin, VEGF; vascular endothelial growth factor, FGF-2; fibrosis growth factor-2; matrix metalloproteinase , (MMP-2,9). The increased levels of interlukins 8 and 17 increase the tumor neoangiogenesis and tumor proliferation. In addition, the extracellular matrix with consist of MMP-2,-9 are considered proteins that assist in th destruction of the cells basal membrane and therefore tumor microinvasion.
and 17 is consequently blocking the neo-angiogenesis and MMP-9 activity, therefore both proliferation and metastatic invasion is reduced (63). At this point we should state that cytokine production can be either the result of a malignancy, or a pre-malignant-inflammatory situation. There is the case as chronic hepatitis where chronic inflammation and cytokine production induce hepatocellular carcinoma. In this situation enhanced neo-angiogenesis and increased activity of MMP-2/-9, assist in tumorigenesis (Figure 1). Therefore targeted treatment modifying the microenvironment could have a place as inhibition of tumorigenesis. Surgically resected patients as in the case of lung cancer patients could have adjuvant treatment at the margins of the surgery stunt, in order to enhance the immune defense mechanisms locally. However; this is stated with caution since an over-stimulation could induce tumorigenesis. Initial experimentation in an in vivo model should provide more data regarding the timing and methodology that should be pursued. In any case the tumor or post surgery microenvironment control should be a future treatment target. There is still a long way of experimentation with IL-8 and IL-17 blocking until a final product is on the market as in the case of IL-6. In conclusion several compounds and methods have been investigated in order to identify which one blocks efficiently the overexpression of these two cytokines. These can be summarized to the following: a) 11-episinulariolide acetate (57), E3330 block of IL-8 (58), Luteolin 8-C-β-fucopyranoside(46), Platinum based chemotherapy more effective in IL-17-positive TICI(65), Ad-si-IL-17 gene therapy(63). These therapeutic approaches have been investigated either by intratumoral injection of intravenous injection. Currently there are antibodies for IL-17 under development and investigated in clinical trials; e.g. vidofludimus in phase IIB for rheumatoid arthritis, psoriasis and inflammatory bowel disease. As of early 2012, clinical trials of IL-17 antibodies against psoriasis Cancer Investigation
IL-8 and IL-17 have shown extremely promising results in Phase II(67). The anti-IL-23 antibody ustekinumab can be used to effectively treat psoriasis by reducing IL-17(68). Raparixin is an inhibitor of IL-8(69). Based on the presented data there are currently dugs on the market blocking the over-expression of IL8 and IL-17, however; off label clinical trials are necessary in order to investigate the effectiveness of these drugs to cancer patients as immunomodulation therapy. DECLARATION OF INTEREST
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The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper. REFERENCES 1. Xie K. Interleukin-8 and human cancer biology. Cytokine Growth Factor Rev 2001;12(4):375–391. 2. Carpagnano GE, Palladino GP, Lacedonia D, Koutelou A, Orlando S, Foschino-Barbaro MP. Neutrophilic airways inflammation in lung cancer: the role of exhaled LTB-4 and IL-8. BMC Cancer 2011;11:226. 3. Araki S, Omori Y, Lyn D, Singh RK, Meinbach DM, Sandman Y, Lokeshwar VB, Lokeshwar BL. Interleukin-8 is a molecular determinant of androgen independence and progression in prostate cancer. Cancer Res 2007;67(14):6854–6862. 4. Gabellini C, Trisciuoglio D, Desideri M, Candiloro A, Ragazzoni Y, Orlandi A, Zupi G, Del Bufalo D. Functional activity of CXCL8 receptors, CXCR1 and CXCR2, on human malignant melanoma progression. Eur J Cancer 2009;45(14):2618–2627. 5. Cheng D, Kong H, Li Y. Prognostic value of interleukin-8 and MMP-9 in nasopharyngeal carcinoma. Eur Arch Otorhinolaryngol 2014;271(3):503–509. 6. Luppi F, Longo AM, de Boer WI, Rabe KF, Hiemstra PS. Interleukin-8 stimulates cell proliferation in non-small cell lung cancer through epidermal growth factor receptor transactivation. Lung Cancer 2007;56(1):25–33. 7. Yu J, Ren X, Chen Y, Liu P, Wei X, Li H, Ying G, Chen K, Winkler H, Hao X. Dysfunctional activation of neurotensin/IL-8 pathway in hepatocellular carcinoma is associated with increased inflammatory response in microenvironment, more epithelial mesenchymal transition in cancer and worse prognosis in patients. PLoS One 2013;8(2):e56069. 8. Kassim SK, El-Salahy EM, Fayed ST, Helal SA, Helal T, Azzam Eel D, Khalifa A. Vascular endothelial growth factor and interleukin8 are associated with poor prognosis in epithelial ovarian cancer patients. Clin Biochem 2004;37(5):363–369. 9. Mayerhofer K, Bodner K, Bodner-Adler B, Schindl M, Kaider A, Hefler L, Zeillinger R, Leodolter S, Joura EA, Kainz C. Interleukin8 serum level shift in patients with ovarian carcinoma undergoing paclitaxel-containing chemotherapy. Cancer 2001;91(2):388– 393. 10. Waugh DJ, Wilson C. The interleukin-8 pathway in cancer. Clin Cancer Res 2008;14(21):6735–6741. 11. Yang G, Rosen DG, Liu G, Yang F, Guo X, Xiao X, Xue F, Mercado-Uribe I, Huang J, Lin SH, Mills GB, Liu J. CXCR2 promotes ovarian cancer growth through dysregulated cell cycle, diminished apoptosis, and enhanced angiogenesis. Clin Cancer Res 2010;16(15):3875–3886. 12. Browne A, Sriraksa R, Guney T, Rama N, Van Noorden S, Curry E, Gabra H, Stronach E, El-Bahrawy M. Differential expression of IL8 and IL-8 receptors in benign, borderline and malignant ovarian epithelial tumours. Cytokine 2013;64(1):413–421. 13. Li Z, Lin SX, Liang YJ, Zong YS. [Effect of macrophage migration inhibitory factor (MIF) on expression of MMP-2,MMP9,and IL-8 in nasopharyngeal carcinoma cell strains]. Ai Zheng 2004;23(2):130–135. C 2014 Informa Healthcare USA, Inc. Copyright
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REVIEW
Interleukin-8 and Interleukin-17 for Cancer Paul Zarogoulidis,1 Fotini Katsikogianni,2 Theodora Tsiouda,3 Antonios Sakkas,4 Nikolaos Katsikogiannis,2 and Konstantinos Zarogoulidis1
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1
Department of Pulmonary, Oncology Unit, “G. Papanikolaou” General Hospital, Aristotle University of Thessaloniki, Thessaloniki, Greece; 2 Department of Surgery (NHS), University General Hospital of Alexandroupolis, Alexandroupolis, Greece; 3 Department of Internal Medicine, “Theageneio” Anticancer Hospital, Thessaloniki, Greece; 4 Department of Pathology, “G. Papanikolaou” General Hospital, Thessaloniki, Greece been associated with advanced stage disease, identified as a prognostic factor (8, 9), and as a possible treatment target through inhibition of IL-8 production and blockage of IL-8 receptors (10, 11). Increased levels have been observed and associated with advanced stage and poor prognosis in cyst fluid, ascites, additionally over-expression of IL-8 has been observed in tumor tissue. The activity of IL-8 is mediated through the receptors IL-8RA and IL-8RB. The first is binding only to IL-8 and granulocyte chemotactic protein-2, while the second with neutrophil activating peptide, granulocyte activation peptide-2, and chemokine Gro-α,β,γ (10). The IL-8RA has been also associated with the pathogenesis of serous tumors (12). IL-8 stimulates proliferation by increasing levels of Cyclin D1 and Cyclin B1 and via activation of Phosphatidylinositide 3-kinases PI3K/Akt and Extracellular signal-regulated kinases Raf/MEK/ERK. Therefore, the mesenchymal transition occurs (13). The invasive capability of a tumor has also been found to be mediated with the increased activation of matrix metalloproteinase-2 and 9 (MMP-2,-9) that are mediated by the levels of IL-8 (14). Therefore, IL-8 levels have been associated with metastatic invasiveness and early recurrence (15). IL-8 expression has been found to be lower in normal cells (10). The MMP as proteolytic enzymes that have the ability to degrade basal membrane. The MMPs that exist in the extracellular matrix play a key role in tumor metastasis. However, although there are 25 MMPs, MMP-9 has been found over-expressed in cancer tissue (16). Further experimentation is overexpression of MMP-9 in a prostate cancer model confirmed the association of MMP-9 with tumor invasiveness (17). It has also been found in a gastric carcinoma model that IL-8 upregulates MMP-9 expression and consequently increased neoangiogenesis is observed (18). IL-17 is a pro-inflammatory cytokine group of ligands that is primarily produced from a subset of CD4+ effector cells known as Th17cells (19). IL17 has been previously associated with the pathogenesis of inflammatory diseases (20). IL-17 induces the expression of
Pro-inflammatory cytokines have been associated with chronic inflammation and inflammatory diseases. Increased levels of interleukins (ILs) have been associated with inflammatory disease exacerbation. ILs levels have been observed to be associated with advance stage cancer for several types of cancer and a poor prognostic maker for malignant disease. Moreover; increased levels of cytokines induce tumorigenesis. There are several paradigms such as the hepatocellular carcinoma induced from chronic inflammation of an underlying hepatitis. In the current review, we will focus on IL-8 and -17. These two ILs as in the case of others, induce neo-angiogenesis through activation of the vascular endothelial growth (VEGF) factor pathway. Additionally, they enhance the activity of matrix metalloproteinase-2 and –9 (MMP-2,-9) which in turn increase the metastatic activity of the underlying malignancy. Inhibition of cytokine production could be a potential treatment both for chronic inflammatory diseases and tumor modulation. Local microenvironment modulation could be applied in surgery resected patients as in the case of lung cancer in order to enhance the local immune activity. Keywords: Interleukin-8, Interleukin-17, Cancer, Immunomodulation
INTRODUCTION Interleukin-8 (IL-8) is a multifunctional pro-inflammatory cytokine produced under inflammation stimulation and is attracting and activating neutrophilic granulocytes (1). Il-8 is secreted by numerous cells such as; (a) mesothelial cells, (b) various tumor cells, (c) neutrophils, (d) endothelial cells, and (e) monocytes (2). It has been characterized as an; angiogenenic factor, motogenic factor, autocrine, and paracrine growth factor. This ability has been observed in numerous cancer types such as; (a) melanoma, (b) prostate cancer, (c) colon cancer, (d) ovarian cancer, (e) nasopharyngeal, (f) lung cancer, and (g) hepatocellular (3–7). IL-8 levels have
Correspondence to: Paul Zarogoulidis, M.D, Ph.D., Pulmonary Department-Oncology Unit, “G. Papanikolaou” General Hospital, Aristotle University of Thessaloniki, Thessaloniki, Greece, email: [email protected] Received 3 January 2014; revised 25 January 2014; accepted 17 February 2014.
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P. Zarogoulidis et al.
chemokines; CXCL1, CXCL6, CXCL8, nuclear factor-kappa B (NF-κB), growth-colony stimulating factor (G-SCF), granulocyte macrophage-colony stimulating factor (GM-CSF), adhesion molecules (ICAM-1), and IL-6 (21, 22). It has been observed that different polymorphisms are associated with different autoimmune diseases such as; rheumatoid arthritis and ulcerative colitis, therefore it has been suggested that the aberrant expression of IL-17 induces abnormal immune response (20). IL-17 has been observed to act upon different cell types (19, 23). The production of IL-17 has been also associated with helicobacter pylori infection and consequently with underlying disease progression (24). IL-17 levels have been found increased in colon (25), ovarian (26), hepatocellular (27), esophageal (28), lung cancer (29), and glioblastoma (30). IL-17 polymorphisms have been associated with gastric cancer through overproduction of IL-17, which in turn over-stimulates the immune system (31, 32). There are controversial data indicating that there is probably a cut off point in the production of IL-17, which if increased it overstimulates the local microenvironment and instead of protecting from mutagenesis it induces it (33). It has been previously observed that there are increased circulating levels of IL-17-producing Th-17 cells in patients with gastric cancer (34). In this subset of patients, colonization with helicobacter pylori does not play a role in gastric cancer development. In colorectal cancer, IL-17 expression was observed to be mediated by ras signaling, implying a new treatment with downregulation of this pathway (35). Furthermore, it has been observed that metastasis is associated with increased levels of IL-17 via activation of CXCR-2-dependent neo-angiogenesis (36). Increased levels of IL-17 have been associated with poor survival and metastasis through increased lymphangiogenesis (37, 38). Increased levels of IL-17 have been found in advance stage disease (29) and could be used as a prognostic marker (39). Increased levels of IL-17 have been associated with early recurrence (40). IL-17 and IL-10 have found elevated in malignant pleural infusion presenting remaining evidence of a mutagenesis process based on inflammation (41). Also in the study by Li et al. (29) it was observed that IL-17 induced metastasis by acting and over-expressing the IL-6 pathway (42, 43). In the current work, we will focus on the clinical data regarding these two pro-inflammatory cytokines. RESEARCH STRATEGIES We performed an electronic article search through PubMed, Google Scholar, Medscape, and Scopus databases, using combinations of the following keywords: IL-8 and cancer, IL-17 and cancer. All types of articles (randomized controlled trials, clinical observational cohort studies, review articles, case reports) were included. Selected references from identified articles were searched for further consideration, without language limitation (Table 1). IL-8 In the study by Shiau et al. (44), it was observed that human papillomavirus (HPV) infection is associated with the activ-
Table 1. Reference Selection Libraries: PubMed, Google Scholar, Medscape, and Scopus databases Key words: interleukin-8 and cancer, interleukin 17 and cancer 1rst Step: 70 studies initially identified and screened 2nd Step: 24 studies excluded by title or abstract examination 3rd Step: 23 studies were identified to be relevant from the references of the 46 previously identified studies 4th Step: 69 references were finally chosen to be included
ity of MMP-2 and -9 that are known to be associated with tumor invasiveness. Moreover; the IL-8 levels were again associated with HPV, neo-angiogenesis, and consequently tumor proliferation. This observation was performed in female never smokers with nonsmall cell lung cancer (NSCLC). In the study by Singh et al. (45), it was observed that IL-8 activity plays a crucial role in breast cancer, it is regulated by Human Epidermal Growth Factor Receptor 2 (HER-2) positive cancers through CXCR1/2 ligands. The IL-8 levels influence the breast cancer stem cell activity by enhancing or downregulating tumorigenesis. Therefore, IL-8 targeted therapy could potentially have a place for breast cancer, however; since it is also regulated by HER-2 expression then conventional therapy could also be sufficient. The activity of MMP-9 and IL-8 were blocked in the study by Park et al. (46) with luteolin 8-C-β-fucopyranoside. In specific, the following pathways were inhibited via ERK/AP-1 and ERK/NF-κB. Metastasis and tumorigenesis were blocked with LUC8C-FP. The polymorphism of IL-8–251 A/T in NSCLC was investigated in a large population study and it was observed that it is not associated with NSCLC. This observation indicates that in specific this expression of pro-inflammatory cytokine is not associated with tumorigenesis (47). However, in the study by Rafrafi et al. (48) high expression of IL-8mRNA was observed in lung cancer patients, which was further associated with tumor size and advance stage disease. In this study the IL-8–251T/T polymorphism was associated with increased tumor size and advance stage lung cancer. Also, this polymorphism was associated with NSCLC adenocarcinoma. In the study by Lattanzio et al. (49) the effect of radiotherapy and anti-angiogenic in the modulation of IL-8 activity was investigated. It is well known that IL-8 increased levels are associated with resistance to anti-angiogenic therapies, this observation was made with bevasizumab therapy. The combination of radiotherapy and antivascular endothelial growth factor (VEGF) reduce effectively the IL-8 levels and its activity. In the study by Yamada et al. (50), biopsies were taken from healthy volunteers and patients with advanced gastric cancer. It was observed that IL-8 mRNA expression level, Helicobacter pylori infection and low PG I/II ratio could be a prognostic cluster without any correlation with cagA mutation. In the study by Browne et al. (12), the levels of IL-8 levels and expression of IL-8RA and IL8-RB were evaluated as a potential prognostic markers for ovarian cancer and risk or tumorigenesis. The IL-8 levels and IL-8RA expression were observed to be potential prognostic biomarkers and were also suggested to be therapeutic targets in ovarian serous carcinoma. In the study by Albulescu et al. (51), the following Cancer Investigation
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IL-8 and IL-17 parameters were investigated in brain cancer: IL-1β, IL-2, IL4, IL-6, IL-8, IL-10, IL-12, GM-CSF, VEGF, Fibrotic growth factor-2 (FGF-2), and TNF-α. The IL-6, IL-10, TNF-α, and IL-1β were upregulated by threefold and VEGF, FGF-2, IL-8, IL-2, and GM-CSF were upregulated by twofold. Therefore, it can be said that in brain cancer strong inflammatory response along with enhanced tumorigenesis is observed. In the study by Jo et al. (52), the blood flow was correlated with IL-8 in order to identify whether it could be correlated with treatment efficiency. The concept is that there are agents targeting Bcl-2 pro-angiogenic pathway that could be used when there is elevated blood flow. In the study by Desai et al.(53), the concept of inhibiting IL-8 was again evaluated as a possible treatment strategy in lung cancer. Data presented again the close association of VEGF with IL-8 levels, moreover; the mesenchymal transition and invasiveness was again associated with IL-8 levels. This observation can be attributed to the MMP-2 and -9 enhanced activity due to the elevated levels of IL-8. The levels of IL-8 and MMP-9 expression were identified as prognostic markers for survival and tumor invasion in nasopharyngeal carcinoma (5). The increased levels of IL-8 induce resistance to anti-VEGF therapy, therefore, a dual therapy of blocking IL-8 and concominately administration of anti-VEGF administration could be potentially an efficient therapy as indicated in the study of Gyanchandani et al.(54). In the study by He et al. (55), tumorigesis was induced with hexavalent chromium Cr (VI) and neoangiogenesis was observed to be correlated with IL-8 leveles and insulin like growth factor-1 (IGF-1). The tumorigenesis of Cr (VI) was regulated by miR-143 expression, by reducing this expression tumorigenesis occurred. In the study by Hsiao et al. (56), it was observed that IL-6 and IL-8 levels are upregulated in the presence of E6 and E7 genes of HPV type 18. Moreover, it was observed that UV radiation increases the activity of E6 and E7 genes. In the study by Hsu et al. (57), it was observed that in epidermal growth factor receptor IL-8 levels and cycloxygenase (COX-2) are overexpressed and downregulated by a coral natural product. The downregulation was achieved through targeting Ca2+ signalingdependent inflammatory diseases. In the study by Cesaratto et al. (58), the APE1 over-expression was associated with chemo-resistance and onset inflammatory response in liver cancer cell lines (Table 2). IL-17 IL-17 family has been associated with numerous inflammatory diseases (20), additionally as it will be described below there, it is a crucial modulator for tumorigenesis and metastasis. In the study by Li et al. (29), it was observed that increased levels of IL-17 were associated with advanced stage disease in lung cancer. IL-7R increased expression was also associated with increased invasiveness. The Signal transducer and activator of transcription 3 (IL-6-STAT3) pathway was activated by the increased levels of IL-7 in order for NSCLC cells to increase their invasiveness. In the study by Petanidis et al. (35), an association of the k-ras status was observed with disease status in colorectal cancer. Moreover, k-ras mutation status was associated with the expression of C 2014 Informa Healthcare USA, Inc. Copyright
interferon-γ (IFN-γ ) and granulocyte macrophage-colony stimulate factor (GM-CSF). K-ras positive patients have the IL-17, IL-22, and 23 differently expressed and the increased levels induce tumorigenesis. The polymorphism G-197A in the IL-17 promoter region was associated with increased gastric cancer risk independently of Helicobacter pylori presence. Moreover, in homozygous patients (AA) at position -197 the chance of developing gastric cancer was 2.9 times higher than single (A) (59). In the study by Wu et al. (40), increased pretreatment levels of IL-17 were associated with poor prognosis of hepatocellular carcinoma and additionally; increased IL-17 levels were associated with early recurrence. In the study by Klimatsidas et al. (41), an association between IL-10, IL-17, and malignant pleural effusion was presented. In specific, it was suggested that the levels measured in the pleural fluid were a result of an inflammatory/mutagenic process. These levels were collated with malignancy type. In the study by Zhang et al. (39), it was observed that TNM stage, positive IL-17, Foxp3 expression, and negative CD4+ were the main prognostic factors. Patients survival undergoing surgery can be more efficiently assessed when combining clinical staging and IL-17/Foxp3 expression. The association of IL-17 expression and levels has been observed in liver fibrosis (60). The IL-6 has been previously also associated with activation of fibrotic mechanisms when cancer patients receive tyrosine kinase inhibitors (TKIs) (42, 43). Since IL-17 uses the IL-6 STAT pathway to enhance the tumor invasion capability it could be stated that these two IL are closely related at least from a clinical point of view. Further investigation of the interaction between these two pathways is needed. The IL-17-producing Foxp3+CD4+ T cells have been observed to play a crucial role in tumorigenesis of colorectal cancer and initiation of chronic bowel disease (61). In the study by Olsson-Akefeldt et al. (62), IL-17 effect on the tumor microenvironment was evaluated and controversial results were presented. Again it seems that the levels of IL-17A play a dual role of enhancing the immune system against tumorigenesis, however; probably after a cut off point that has not been established until now they induce tumorigenesis. In the study by Hayata et al. (63), the effect of IL-17 was again evaluated in the tumor microenvironment by administration of gene therapy. It was observed that at the sites of IL-17 inhibition the tumor proliferation was blocked through down-regulating neo-angionenesis and regulation of cytotoxic T lymphocytes. The MMP-9 activity that is a protein that enhances the cancer cell invasion and metastasis was also blocked with local control of IL-17. In the study by Jordan Wiliams et al. (64), the basic leucine zipper transcription factor (BATF) that dimerizes with the JUN proteins to direct patterns of activator protein-1 (AP-1)mediated gene expression was investigated. In specific, BATF was investigated how it interacts with the regulation of IL-17A and the results indicated that it could be used as a treatment target. IL-17 was also investigated in esophageal squamous cell carcinoma as a prognostic factor (28). It was observed that mast cells play a crucial role in tumor immunity by releasing IL-17 in muscularis propia and therefore
Colorectal patients
IL-17 G-197A polymorphism (patients) Hepatocellular
Malignant pleural fluid Lung cancer
Liver fibrosis Dendritic cells
Colon
Mice expressing human HA-tagged BATF Esophageal squamous cell carcinoma
Glioblastoma (patients) Colon cancer
Ovarian (patients)
Hepatocellular (Patients)
Breast (patients) Ovarian (patients)
Lung cancer
Breast cancer NSCLC Uveal melanoma
Petanidis et al.
Rafiei et al.
Klimatsidas et al. Zhang et al.
Du et al. Olsson-Akefeldt et al.
Hayata et al.
Jordan-Williams et al. Wang et al.
Cui et al. Straus D.
Lan et al.
Liao et al.
Chen et al. Droeser et al.
Shiau et al.
Singh et al. Bhat et al. Lattanzio et al.
Wu et al.
NSCLC patients
Cancer
Li et al.
Author
Table 2. Clinical Studies
ELISA, CM and Flow cytometry, tissue microarray design and immunocytochemistry IHC, quantification of staining results Tissue microarray construction, IHC, validation analysis of IL-17 staining Zymography, Western blot, RNA extraction and RT-PCR, ELISA Review DNA extraction, PCR, ELISA, western blot
IHC, immunofluorescence, immunoelectron, quantification of immunofluorescence, evaluation of immunohistology IHC, Glucose and L-lactate assays, Growth factors assays, Protein experiments and AKT phosphorylation assays. IHC, selection of the cut-off point for expression of IL-17
ELISA, RT-PCR, IHC TRAP and Hoechst staining, CFSE and CD14-PE labeling for proliferation study, Flow cytometry, western blot, immunocytofluorescence labeling, affymetrix genechip study, RT-PCR, Functional analysis Cytotoxic assay, flow cytometry analysis, IHC and quantitative microscopy, Measurement of cytokine and chemokine content in tumor tissues and in vitro ELISA, Flow Cytometry
ELISA IHC, Eli Vision two-step kit
Quantification of serum levels of cytokines, FACS
DNA-RNA extraction, ELISA, RT-PCR, Westen blot, IHC, MTT cell viability, SPOT GM-CSF-IFN-γ , TUNEL assay, K-ras status IL-17 genotyping, Helicobacter pylori detection
Invasive assay, Plasmid construction, western blotting, evaluation of tumor progression in vivo
Method of Evaluation
Results
IL-8 levels associated with neo-angiogenesis, MMP-2 and -9 associated with tumor invasiveness-metastasis IL-8 levels regulate breast cancer stem cell activity. IL-8–251 A/T IL-8 levels and signaling were more effectively reduced when combination of radiotherapy and bevacizumab were administered
IL-17 is a good prognostic marker for glioblastoma TNF-α and IL-17 increased levels enhance tumorigenesis in colorectal cancer IL-17 and TAM density are correlated. IL-17 levels are associated with PFS and survival IL-17 and IL-17RE increased levels are a poor prognostic factor for HCC
IL-17 increased in propia muscularis along with mast cells, macrophages and CD8+ T cells
IL-17 inhibition with gene therapy down regulates neo-angiogenesis and MMP-9 activity, therefore inhibiting tumorigenesis BTAF could be a possible treatment target
G-197A polymorphism associated with development of gastric cancer Increased pretreatment levels associated with poor prognosis and early recurrence IL-10, IL-17 levels associated with malignancy TNM staging, IL-17 and Foxp3 expression, CD4+negative expression associated with disease prognosis IL-17 levels associated with liver fibrosis Modulation of microenvironment
IL-17 levels associated with advance stage, IL-17R increased expression associated with increased invasiveness. K-ras positive mutation associated with increased tumorigenesis
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(45) (47) (49)
(44)
(37) (65)
(27)
(26)
(30) (25)
(14)
(64)
(63)
(60) (62)
(41) (39)
(40)
(59)
(35)
(29)
Ref
P. Zarogoulidis et al.
Cancer Investigation
C 2014 Informa Healthcare USA, Inc. Copyright
Gastric cancer
Ovarian cancer
Breast cancer
Brain cancer (patients)
Head and Neck squamous cell carcinoma (patients) Lung cancer
Yamada et al.
Browne et al.
Park et al.
Albulescu et al.
Jo et al.
Lung cancer
Basal cancer
Lung cancer
Liver cancer cell lines
Gyanchandani et al.
He et al.
Hsiao et al.
Hsu et al.
Cesaratto et al.
MTT and cell growth assays, MTS assay, Nile red staining, western blot, immunofoulorescence assay and confocal analysis, RT-PCR, soluble cytokines determination, transient transfection experiments
ELISA, RT-PCR, western blotting, luciferase assay
ELISA, antibody array, transwell migration, cell proliferation, western blot, capillary-tube formation, IHC and immunofluorescence RT-qPCR, colony isolation, tube formation assay, IHC, miRNA transfection, chorioallantoic membrane assay, miRNA luciferase reporter constructs and luciferase activity assay Editorial
Cell adhesion assay, confocal immunofluorescent staining, western blotting and IP, motility and invasion assay, gelatin zymography, anoikis assay, cell transfection, soft agar colony formation assay Serum IL-8 and MMP-9 assay
CT perfusion, serum samples and ELISA
MTS assay, wound healing assay, matrigel invasion assay, gelatin zymography, RT-PCR, ELISA, western blotting xMAP analysis and ELISA,
IHC, assessment of expression
Determination of mRNA expression, genotyping of cytokine gene polymorphisms RT-PCR, genotyping, antibody test
UV increase the activity of E6 and E7 genes in HPV type 18 virus and IL-6 and IL-8 levels are increased in the presence of E6 and E7 Coral natural product a potential treatment for increased levels of COX-2 and IL-8 APE1 responsible for chemo-resistance
miR-143 regulates tumorigenesis from Cr (VI) and IL-8 levels are associated with neoangiogenesis
IL-8 increased levels and MMP-9 expression are considered as prognostic markers IL-8 and anti-VEGF co-administration more efficient than one agent alone
Helicobacter Pylori, mRNA expression and low PG I/II ratio are associated with increased rick of gastric cancer IL-8 and IL-8RA potential prognostic markers and treatments LUC8C-FP a possible treatment for blocking metastasis and tumorigenesis IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, GM-CSF, VEGF, FGF-2 and TNF-α. The IL-6, IL-10, TNF-α, IL-1β were up-regulated by threefold and VEGF, FGF-2, IL-8, IL-2, and GM-CSF were upregulated by twofold Elevated blood flow associated possibly with effective Bcl-2 targeted treatment IL-8 possible targeted treatment which down-regulates the VEGF pathway
IL-8–251T/T association with NSCLC
(58)
(57)
(56)
(55)
(54)
(5)
(53)
(52)
(51)
(46)
(12)
(50)
(48)
NSCLC; nonsmall cell lung cancer, IL-17; interleukin-17, IL-17R; interleukin 17 receptor, ELISA; Enzyme-Linked Immunosorbent Assay, RT-PCR; Reverse transcription polymerase chain reaction, IHC; immunohistochemistry, MTT; tetrazolium dye, GM-CSF; granulocyte macrophage-colony stimulating factor, IFN-γ ; interferon-γ , TUNEL; Terminal deoxynucleotidyl transferase dUTP nick end labeling, SPOT; enzyme-linked immunosorbent spot, FACS; Fluorescence-activated cell sorting, CFSE; Carboxyfluorescein succinimidyl ester (CFSE) is a fluorescent cell staining dye, MMP-9; matrix metalloproteinase, TNF-α; tumor necrosis factor-α, TAM; tumor associated macrophages, PFS; progression free survival, HCC; hepatocellular carcinoma, CM; solid-phase cytometry, MTS; MTT, CT; computed tomography, xMAP; multiplex assay, VEGF; vascular endothelial growth factor, UV; ultra violet radiation.
Nasopharyngeal cancer (Patients) Head and neck cancer
Cheng et al.
Desai et al.
Lung cancer (patients)
Rafrafi et al.
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IL-8 and IL-17
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P. Zarogoulidis et al.
could serve as a prognostic factor for squamous cell carcinoma. Activated macrophages and increased levels of density effector CD8+ T cells were also observed in muscularis propia along with increased levels of IL-17. In the study by Cui et al. (30), the expression of IL-17 was evaluated in glioblastoma cells and was observed to be a valuable prognostic marker. In specific, higher levels of IL-17 were correlated with progression free survival (PFS), and death rate. The IL-17 and PFS were also observed to be independent factors affecting the overall survival. Therefore, it can probably be speculated that in the case of glioblastoma the IL-17 is modulating the tumor microenvironment as a defense mechanism. The same observation was made for ovarian cancer, increased levels of IL-17 were associated increased PFS and survival (26). Furthermore, it was observed that the density of tumor-associated macrophages (TAMs) was associated with the levels of IL-17. Moreover, increased levels of IL-17 have been associated with chemo-sensitivity to platinum-based chemotherapy (65). The contribution of tumor necrosis factor-a and IL-17 in colon cancer tumorigenesis was investigated, and it was observed that these pro-inflammatory cytokines contribute to cancer proliferation through stimulation of glycolysis and growth factor production (25). This study by Straus D. indicates again that IL-17 blockage could be a future treatment proposal. In the study by Liao et al. (27) the IL-17 and IL-17RE were found to be poor prognostic factors for hepatocellular carcinoma. In this type of cancer the chronic nature of the inflammation and continuous activity of IL-17 producing CD4+ T cells could be the substrate of tumorigenesis. Targeted therapy in a form of pulse adjuvant treatment could be helpful, activating the defense mechanisms but not over-expressing them. Moreover; increased IL-17 levels were associated with poor survival in breast cancer (66). Increased levels of IL-17 were also associated with HER-2, advanced T stage and presented shorter disease free survival when combined (Table 2). DISCUSSION Cytokines have been observed increased in several chronic inflammatory diseases, and increased levels have been associated with disease exacerbation. IL-8 has been extensively investigated in lung cancer, while IL-17 has been investigated in a variety of cancer types. There is the case of crohn‘s disease were treatment with monoclonal antibodies inhibiting the inflammation cascade and cytokine production have provided an efficient disease control for many patients (42, 43). Inflammatory cytokines have been observed increased in advanced stage disease for many types of cancer and increased levels are a poor prognostic marker. The IL-17 family consists of numerous subdivisions such as; A, B, C, D, E and F. The IL-17A blocking has been tested for autoimmune disorders and could be a future treatment methodology. Regarding B, C, D and E their function still remain to be elucidated. IL-17F plays a crucial role in host defense from bacteria (20). The modulation of the tumor microenvironment is possible through inhibition of IL-17 activity. The inhibition of IL-8
Figure 1. IL; interleukin, VEGF; vascular endothelial growth factor, FGF-2; fibrosis growth factor-2; matrix metalloproteinase , (MMP-2,9). The increased levels of interlukins 8 and 17 increase the tumor neoangiogenesis and tumor proliferation. In addition, the extracellular matrix with consist of MMP-2,-9 are considered proteins that assist in th destruction of the cells basal membrane and therefore tumor microinvasion.
and 17 is consequently blocking the neo-angiogenesis and MMP-9 activity, therefore both proliferation and metastatic invasion is reduced (63). At this point we should state that cytokine production can be either the result of a malignancy, or a pre-malignant-inflammatory situation. There is the case as chronic hepatitis where chronic inflammation and cytokine production induce hepatocellular carcinoma. In this situation enhanced neo-angiogenesis and increased activity of MMP-2/-9, assist in tumorigenesis (Figure 1). Therefore targeted treatment modifying the microenvironment could have a place as inhibition of tumorigenesis. Surgically resected patients as in the case of lung cancer patients could have adjuvant treatment at the margins of the surgery stunt, in order to enhance the immune defense mechanisms locally. However; this is stated with caution since an over-stimulation could induce tumorigenesis. Initial experimentation in an in vivo model should provide more data regarding the timing and methodology that should be pursued. In any case the tumor or post surgery microenvironment control should be a future treatment target. There is still a long way of experimentation with IL-8 and IL-17 blocking until a final product is on the market as in the case of IL-6. In conclusion several compounds and methods have been investigated in order to identify which one blocks efficiently the overexpression of these two cytokines. These can be summarized to the following: a) 11-episinulariolide acetate (57), E3330 block of IL-8 (58), Luteolin 8-C-β-fucopyranoside(46), Platinum based chemotherapy more effective in IL-17-positive TICI(65), Ad-si-IL-17 gene therapy(63). These therapeutic approaches have been investigated either by intratumoral injection of intravenous injection. Currently there are antibodies for IL-17 under development and investigated in clinical trials; e.g. vidofludimus in phase IIB for rheumatoid arthritis, psoriasis and inflammatory bowel disease. As of early 2012, clinical trials of IL-17 antibodies against psoriasis Cancer Investigation
IL-8 and IL-17 have shown extremely promising results in Phase II(67). The anti-IL-23 antibody ustekinumab can be used to effectively treat psoriasis by reducing IL-17(68). Raparixin is an inhibitor of IL-8(69). Based on the presented data there are currently dugs on the market blocking the over-expression of IL8 and IL-17, however; off label clinical trials are necessary in order to investigate the effectiveness of these drugs to cancer patients as immunomodulation therapy. DECLARATION OF INTEREST
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