IL-6与VEGF在食管鳞状细胞癌中的表达及其临床意义

目的：检测食管鳞状细胞癌（esophageal squamous cell cancer,ESCC）患者血清中白细胞介素-6（interleukin-6,IL-6）和ESCC组织中血管内皮生长因子（vascular endothelial growth factor,VEGF）的表达及其两者关系,并研究两者表达对ESCC的临床意义。方法：收集河北医科大学第四医院胸外科于2014年1月至2015年1月期间行ESCC切除术的患者52例,每例患者均取原发灶组织和癌旁组织标本;同时在术前抽取患者外周血5 ml,再取52例健康体检者外周血5 ml为血清对照。应用ELISA法测定ESCC患者血清中IL-6的水平,免疫组化技术检测VEGF在ESCC组织中的表达,RT-PCR法检测肿瘤组织中IL-6和VEGF mRNA的表达情况。结果：ESCC患者血清IL-6表达水平为（116.71±25.98）pg/ml,明显高于健康对照组的\[（78.43±9.36）pg/ml\]（P＜0.05）,血清中IL-6的表达水平与患者肿瘤分化程度、TNM分期、肿瘤浸润深度和淋巴结转移相关（P＜0.05）。肿瘤组织VEGF蛋白阳性表达率明显高于癌旁组织（67.31% vs 32.69%,P<0.01),且与患者肿瘤分化程度、TNM分期、肿瘤浸润深度和淋巴结转移相关（P＜0.05）。肿瘤组织中IL-6 mRNA的表达与VEGF mRNA的表达呈显著正相关（r=7.113,P＜0.05）。结论：ESCC患者肿瘤组织中IL-6和VEGF均呈高表达且两者呈正相关,两者可能在ESCC侵袭和转移中发挥重要作用。     

Hepatocyte growth factor/scatter factor-induced activation of MEK and PI3K signal pathways contributes to expression of proangiogenic cytokines interleukin-8 and vascular endothelial growth factor in head and neck squamous cell carcinoma

The proangiogenic activity of hepatocyte growth factor (HGF)/scatter factor has been closely associated with its ability to stimulate endothelial cell chemotaxis, migration, proliferation, and capillary formation. However, the potential of HGF as a paracrine factor in regulating the expression of angiogenesis factors by tumor cells is not widely appreciated. We observed that increased HGF was correlated with higher levels of angiogenesis factors interleukin (IL)-8 and vascular endothelial growth factor (VEGF) in serum of patients with head and neck squamous cell carcinoma (HNSCC) as compared with that in normal volunteers and hypothesized that HGF may regulate angiogenesis factor production by tumor cells through the activation of its receptor c-Met, which is expressed by HNSCC cells. To test this hypothesis, we examined the effect of HGF treatment on IL-8 and VEGF expression by a panel of primary keratinocytes and HNSCC lines. HGF induced a significant dose-dependent increase in IL-8 and/or VEGF cytokine production in eight HNSCC lines tested, which is not observed in normal keratinocytes. In addition, HGF increased mRNA expression of IL-8 in 3 of 6 and VEGF in 5 of 6 HNSCC lines. The increase in induction of these factors by HGF corresponded to an increase in phosphorylation of c-Met in HNSCC. HGF-induced phosphorylation of mitogen-activated protein/extracellular signal-regulated kinase kinase (MEK) pathway substrate p42/p44(erk) and phosphatidylinositol 3'-kinase (PI3K) pathway substrate Akt provided evidence for downstream activation of MEK and PI3K pathways in HNSCC. Inhibitors of MEK (U0126) and PI3K (LY294002) blocked p42/p44(erk) and Akt, respectively, and partially blocked HGF-induced production of IL-8 and VEGF, whereas the combination of U0126 and LY294002 completely inhibited expression of IL-8 and VEGF by UMSCC-11A. Our results demonstrate that HGF can promote expression of angiogenesis factors in tumor cells through both MEK- and PI3K-dependent pathways. Understanding HGF/Met paracrine regulatory mechanisms between tumor and host cells may provide critical information for targeting of therapies against angiogenesis.     

Macrophage migration inhibitory factor: roles in regulating tumor cell migration and expression of angiogenic factors in hepatocellular carcinoma

Macrophage migration inhibitory factor (MIF) may contribute to multiple aspects of tumor progression, including control of cell proliferation, differentiation, cell survival and angiogenesis. However, the potential roles of MIF in regulating hepatocellular carcinoma (HCC) tumor cell migration and the expression of angiogenic factors by HCC tumor cells have not been studied yet. In our study, we reported that intracellular MIF mRNA and protein were overexpressed in HCC tissues compared to nontumor tissues by using in situ hybridization and immunohistochemic staining. HCC tumor cell lines also secreted large amounts of MIF into the supernatants of tumor cell culture. To assess the role of MIF in HCC, we employed the transwell invasion chamber to study the effect of MIF on tumor cell migration. Our results showed that recombinant MIF and the supernatants of tumor cell line culture could enhance the invasion and migration of HCC cells. This effect can be inhibited by the addition of a neutralizing anti-MIF antibody. We observed that increased MIF serum levels correlated with higher levels of interleukin-8 (IL-8) in the sera of patients with HCC than in normal volunteers. We therefore hypothesized that MIF may regulate the production of angiogenic factors by HCC cells. To test this hypothesis, we examined the effect of MIF treatment on vascular endothelial growth factor (VEGF) and IL-8 expression by HCC cell lines. MIF induced a significant dose-dependent increase in IL-8 and VEGF production. Taken together, our results indicated that MIF may act as an autocrine-acting factor that stimulates angiogenesis and metastasis in HCC by promoting expression of angiogenic factors and migration of tumor cells. A more detailed understanding of the MIF regulatory mechanisms involved may provide insight into new direction in the treatment of HCC.     

Clinical significance of serum soluble interleukin 2 receptor-alpha in esophageal squamous cell carcinoma.

Although the serum level of soluble interleukin-2 receptor alpha (sIL-2Ralpha) has been shown to correlate with progression and prognosis of several cancers, data to support its clinical significance to esophageal squamous cell carcinoma (ESCC) are limited. This study was conducted to assess the prognostic value and source of sIL-2Ralpha in patients with ESCC. From January 1986 to June 1997, 125 patients with histopathologically confirmed ESCC were enrolled for study. Ninety-three patients underwent en bloc esophagectomy, and 32 patients with unresectable tumor underwent palliative surgery. Four (4.3%; 4 of 93) patients died of surgical complications. Serum levels of sIL-2Ralpha were measured by ELISA. Expression of IL-2Ralpha, IL-2Rbeta, and IL-2Rgamma in the pathological section was determined, respectively, by immunohistochemistry (IHC) and in situ hybridization (ISH). Compared with the healthy control group (1020 +/-476 pg/ml, n = 103), ESCC patients tended to have significantly higher serum sIL-2Ralpha concentrations (1424 +/- 798 pg/ml, n = 121). The sIL-2Ralpha level was correlated with age, Tumor-Node-Metastasis classification, tumor stage, reading score of the IHC staining, and survival but not with the pathological grade or lymphovascular invasion. Prognosis was worse for patients with high sIL-2Ralpha levels (> or =1500 pg/ml) than for those with low serum sIL-2Ralpha levels (< 1500 pg/ml; P = 0.0209). It can be used as an independent prognostic factor of ESCC. In the pathological sections, expression of IL-2Ralpha, IL-2Rbeta, and IL-2Rgamma was detected in 17 (18.1%), 83 (89.2%), and 83 (89.2%) cases, respectively, by IHC, and the message of IL-2Ralpha was identified in tumor cells by ISH in 30.1% (28 of 93) of the cases. Serum concentrations of sIL-2Ralpha are frequently elevated in ESCC patients and are correlated with disease progression and survival. These data indicate that, in addition to activated T cells, cancer cells could be an important source of sIL-2Ralpha in ESCC patients.     

Serum vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) levels in small cell lung cancer

This study was conducted to determine the value of the angiogenic serum factors, vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8), in patients with small cell lung cancer (SCLC). These serum angiogenic factors were measured of 34 SCLC patients on the before and after chemotherapy in comparison with 20 healthy controls using ELISA method. Serum levels of VEGF and IL-8 were significantly increased in SCLC patients compared with healthy controls (p < 0.001). No statistically significant relationships was found between investigated elevated serum angiogenic parameters and various characteristics of patients and disease such as disease stage and tumor burden. Likewise, we also found no correlation between serum angiogenic factors. Cytotoxic therapy of patients was accompanied by unchanged serum levels of angiogenic factors. Contrary to serum IL-8, elevated serum levels of VEGF was determined as a prognostic factor for survival by univariate analysis (p = 0.05). Multivariate analysis revealed that independent prognostic factors of overall survival included only response to chemotherapy and weight loss (p < 0.001 for both). In conclusion, our data suggest that the angiogenic serum factors, VEGF and IL-8, are useful diagnostic factors, but not predictive and prognostic markers for overall survival in SCLC patients.     

Cytokines facilitate chemotactic motility of breast carcinoma cells

BACKGROUND: Both growth and motility of various tumor cells have been shown to be influenced by surrounding cells such as lymphocytes, histiocytes and fibroblasts through various cytokines, growth factors and extracellular matrices. The role of cytokines and extracellular matrices produced by lymphocytes, histiocytes and fibroblasts on migration and invasion of breast carcinoma cells has not been fully investigated METHODS: We investigated the effect of hepatocyte growth factor (HGF), interleukin (IL)-6, IL-8, IL-11, soluble type IV collagen and soluble laminin on the migration of 3 human breast carcinoma cell lines, MDA-MB-231, MCF-7 and T-47D, using a cell culture insert and a biocoat matrigel invasion chamber to assess migration across a matrigel-coated polyethylene telephtalate membrane. RESULTS: HGF, IL-6, IL-11 and IL-8 induced significant migration of MDA-MB-231 cells depending on the dose of each cytokine. However, type IV collagen and laminin inhibited migration of MDA-MB-231 cells. In contrast, IL-8 inhibited migration of MCF-7 cells and IL-6 induced significant migration of T-47D cells, while no other cytokine or extracellular matrix induced significant migration of MCF-7 and T-47D cells. Only HGF induced significant invasion of MDA-MB-231 cells depending on the dose. MCF-7 and T-47D cells did not invade in response to any of the cytokines and extracellular matrices tested. CONCLUSIONS: These results suggest the possibility that the potency of chemotaxis or chemoinvasion differs according to the breast carcinoma cell line and that various cytokines and extracellular matrices secreted by lymphocytes, histiocytes and fibroblasts in the stroma of breast carcinoma can affect the invasion of breast carcinoma cells.     

ALC1蛋白在食管鳞癌中的表达及其对细胞增殖?侵袭?迁移的影响

  目的  探讨ALC1（amplified in liver cancer 1）在食管鳞癌组织中的表达及与临床病理特征及总生存率关系,检测过表达ALC1基因对食管癌细胞恶性生物学行为的影响。  方法  采用免疫组织化学方法检测245例食管鳞癌组织及癌旁组织中ALC1蛋白的表达,并探讨其与食管鳞癌患者性别、年龄、分化程度、浸润深度、TNM分期、远处淋巴结转移关系及总生存率关系;采用MTT法、克隆形成实验、Transwell实验及细胞划痕实验等观察高表达ALC1基因在食管癌细胞中的增殖、侵袭及迁移作用。  结果  ALC1蛋白在食管癌组织中的阳性表达率明显高于癌旁组织（41.6% vs. 21.2%,P < 0.05）;ALC1的高表达与肿瘤的浸润深度、TNM分期和淋巴结转移明显相关（P < 0.05）。ALC1高表达的食管鳞癌患者总生存率低。ALC1基因能够促进KYSE30食管癌细胞过度增殖、侵袭和迁移。  结论  ALC1表达升高可能与食管鳞癌的发生、发展相关,导致总生存率下降,高表达的ALC1基因增强KYSE30食管癌细胞的增殖、侵袭及迁移能力,检测ALC1可能为食管癌预后判断提供依据。      

Overexpression of macrophage migration inhibitory factor induces angiogenesis in human breast cancer

Macrophage migration inhibitory factor (MIF) is known to be an important contributor to tumor progression. Overexpression of MIF has been reported in different types of tumors. However, the correlation between MIF expression and tumor pathologic features in patients with breast cancer has not been elucidated. In this study, we examined the expression of MIF, vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) in human tissues with or without tumor. In addition, we investigated the expression of MIF in MDA-MB-231, MCF-7 (breast cancer cell lines) and MCF-10A (epithelial cell line) cells, and its effect on VEGF and IL-8. We found that MIF was overexpressed in breast cancer tissues compared with normal ones. The level of MIF showed the positive correlation between the expression of IL-8 and tumor microvessel density (MVD). The patients with positive MIF expression in tumor tissues showed a significantly worse disease-free survival compared with negative ones. Increased MIF serum levels were also found to correlate with higher levels of IL-8 in the sera of the patients with breast cancer. In vitro experiments successfully detected MIF in breast cell lines. However, the expression level of it by normal epithelial cells was much less than that of cancer cells. Exogenous MIF did not cause endothelial tube formation and migration but induced a dose dependent increase in VEGF and IL-8 secretion in breast cancer cell lines. In summary, our studies show that human breast cancer tissue expresses MIF. Its in vitro effect on VEGF and IL-8 indicates that MIF may contribute to tumor in angiogenesis and thus play an important role in the pathogenesis of breast cancer.     

Interleukin-18 is a critical factor for vascular endothelial growth factor-enhanced migration in human gastric cancer cell lines

Cell migration and angiogenesis are key steps in tumor metastasis. However, the mechanism of migration regulated by vascular endothelial growth factor (VEGF), a potent regulator of angiogenesis, is not completely understood. This study examined the relationship between VEGF and migration, along with the mechanism involved in the VEGF-regulated migration of human gastric cancer cells. The level of cell migration was increased by recombinant human (rh)VEGF-165 in the VEGF receptor-2-expressing SNU-601 cells. Interleukin (IL)-18 is associated with the malignant progression of tumors. Accordingly, this study examined the effect of IL-18 on the migration of cancer cells in order to identify the factors involved in VEGF-enhanced migration. Inhibiting IL-18 markedly reduced the level of VEGF-enhanced migration, and IL-18 increased cell migration directly through filamentous-actin polymerization and tensin downregulation. It was confirmed that rhVEGF-165 increased IL-18 production significantly. An antioxidant and an extracellular signal-regulated kinase (ERK)1/2-specific inhibitor blocked rhVEGF-165-enhanced IL-18 production. Accordingly, rhVEGF-165 increased the generation of region of interest (ROI) and activated the ERK1/2 pathway. These results suggest that rhVEGF-165 enhances IL-18 production via the generation of ROI and ERK1/2 phosphorylation, which results in the increased migration of gastric cancer cells.     

Predicting treatment responses and disease progression in myeloma using serum vascular endothelial growth factor and hepatocyte growth factor levels

Angiogenesis is a crucial process in the progression of multiple myeloma (MM). Vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) are multifunctional cytokines that potently stimulate angiogenesis during tumor neovascularization. VEGF is secreted by MM cells. It induces proliferation of MM cells and stimulates IL-6 expression by microvascular endothelial cells and bone marrow stroma cells, suggesting both autocrine and paracrine functions for VEGF in MM. HGF and the HGF receptor, c-Met, are expressed simultaneously in MM cell lines and in freshly isolated MM cells, suggesting a possible role for HGF in MM cell proliferation. This review focuses on the clinical significance of serum levels of VEGF and HGF in MM.     

Clinical significance of osteopontin in esophageal squamous cell carcinoma: comparison with common tumor markers

OBJECTIVE: Osteopontin (OPN) is a secreted integrin-binding glycophosphoprotein that may have a role in head and neck squamous cell carcinoma (SCC). To evaluate the clinical significance of OPN in esophageal squamous cell carcinoma (ESCC), we compared plasma OPN levels with those of common tumor markers. METHODS: Preoperative plasma OPN levels were measured by enzyme immunoassay in 103 ESCC patients. Serum SCC antigen, Cyfra 21-1, and carcinoembryonic antigen (CEA) levels were also measured routinely at admission by radioimmunoassay. RESULTS: Plasma OPN levels ranged from 82.8 to 1,980 ng/ml. High OPN level was associated with lymph node metastasis (p = 0.05), but not with tumor histology or depth of invasion. The overall survival of the patients with high OPN levels was worse than that of those with low OPN levels (p = 0.02). SCC antigen and Cyfra 21-1 levels were associated with the depth of tumor invasion, the tumor diameter, lymph node metastasis, and the overall survival, but CEA was not associated with these clinicopathological factors. Combined evaluation of OPN plus Cyfra 21-1 or OPN plus SCC antigen was useful as an independent prognostic indicator. CONCLUSION: Measurement of the plasma OPN level, as well as serum SCC antigen and Cyfra 21-1, may help to predict the progression of ESCC.     

Pretreatment serum levels of cytokines and cytokine receptors in patients with non-small cell lung cancer, and correlations with clinicopathological features and prognosis. M-CSF - an independent prognostic factor

OBJECTIVES: Cytokines are potential new serum markers, especially desirable for malignancies with poor prognosis like non-small cell lung cancer (NSCLC). METHODS: Cytokines, tumor necrosis factor alpha (TNFalpha), interleukin (IL)-6 and IL-8, soluble TNF (sTNF) RI, sTNF RII, soluble IL-2 receptor-alpha, IL-1 receptor antagonist (IL-1ra), IL-10, vascular endothelial growth factor, basic fibroblast growth factor, and macrophage (M-CSF) and granulocyte colony-stimulating factor, as well as tumor markers - carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC) and CYFRA 21.1 - were assessed in the sera of 103 untreated NSCLC patients, and these cytokines and tumor markers were referred to clinical parameters of the disease and to the overall survival of patients evaluated during a 6-year follow-up. RESULTS: Most of the factors analyzed were found to be elevated in the sera of NSCLC patients, and increases in IL-6, IL-8 and sTNF RI were noted in the greatest proportion of stage I patients. Most cytokine/cytokine receptor levels revealed higher sensitivity than the standard tumor markers; IL-6 and IL-1ra levels were significantly different in patients with squamous cell versus adenocarcinoma; IL-6 and IL-10 were related to the tumor size, while IL-6 and M-CSF levels significantly increased with disease progression. A significant prognostic value of pretreatment serum M-CSF and CEA levels in NSCLC patients has been shown, but only M-CSF proved to be an independent prognostic factor. CONCLUSIONS: Increased pretreatment serum M-CSF level is a significant independent predictor of poor survival in patients with NSCLC.     

The role of circulating IL-8 and VEGF protein in the progression of gastric cancer

Both vascular endothelial growth factor (VEGF) and interleukin 8 (IL-8) play an important role in the progression of gastric cancer (GC). In this study, we investigated whether circulating levels of VEGF or IL-8 in drainage veins of GC patients were correlated with any clinicopathological factors. Thirty-seven patients with primary GC who underwent gastrectomy at our department between 1999 and 2002 were analyzed. Blood samples were drawn from a peripheral vein just before surgery and from a drainage vein immediately after laparotomy. Plasma VEGF levels were significantly higher than those in 10 healthy controls. There was no correlation between VEGF levels in drainage veins and any clinicopathological variable, whereas there was a significant relationship in the case of VEGF levels in peripheral veins; the levels were higher in patients with venous invasion. We found a significant relationship between IL-8 levels in drainage veins and both tumor size and lymph node metastasis, whereas no significant relationship between IL-8 levels in peripheral veins and any variable was found. There was no correlation between VEGF and IL-8 levels in drainage veins. Large tumors, deeply invasive tumors, lymph node involvement, venous invasion and high IL-8 levels in drainage veins were all significantly associated with shorter disease-free survival, although multivariate analysis revealed that lymph node involvement was the only independent prognostic factor. In conclusion, the measurement of IL-8 levels in drainage veins of GC patients may reflect production mainly by the primary lesion and is valuable as an indicator of risk for recurrent disease.     

Interleukin-6, interleukin-10, and vascular endothelial growth factor in metastatic renal cell carcinoma: prognostic value of interleukin-6--from the Groupe Francais d'Immunotherapie.

PURPOSE: Few clinical prognostic factors have been identified for patients with metastatic renal cell carcinoma (MRCC), and no biomarker is known in this disease. Several endogenous cytokines have demonstrated interesting and significant correlations with survival in these patients. Our objective was to analyze the prognostic value of circulating vascular endothelial growth factor (VEGF), interleukin-10 (IL-10), and interleukin-6 (IL-6). PATIENTS AND METHODS: Serum levels of IL-6, IL-10, and VEGF were measured in patients with MRCC. Their prognostic value for response to treatment and progression-free and overall survival was evaluated. Pretreatment samples were obtained from 138 patients of a large randomized multicentric trial. Endogenous cytokine levels were determined using immunoassays. Univariate and multivariate analyses were performed to evaluate the prognostic value of each factor further controlled by an internal validation test. Threshold values for serum IL-6 and VEGF were determined using the quartile method. RESULTS: Serum IL-6 was detectable in 70% of the patients. IL-10 and VEGF were elevated in 8% and 71% of the patients, respectively. None of these circulating factors was correlated with response to treatment. IL-10 was not significantly correlated with progression-free or overall survival. Despite significant correlation with survival, VEGF was not an independent prognostic factor in the multivariate analysis. Finally, IL-6 was significantly correlated with progression-free survival and overall survival, and has prognostic value for overall survival. CONCLUSION: Circulating IL-6 level appears to be an important independent prognostic factor in patients with MRCC; if confirmed in further studies, it could be considered for treatment decisions in these patients.     

Matrix Metalloproteinase-13 - A Potential Biomarker for Detection and Prognostic Assessment of Patients with Esophageal Squamous Cell Carcinoma

Background: Matric metalloproteinase (MMP) 13 gene expression is increased in esophageal squamous cell carcinomas (ESCCs) and associated with increasing tumor invasion, lymph node involvement and decreased survival rates. Levels of the circulating enzyme may be elevated and used as a marker of tumor progression. In this study, clinical application of MMP-13 serum levels was evaluated for early detection, prediction of prognosis and survival time of ESCC patients. Materials and Methods: Serum levels of MMP13 were determined by ELISA in 66 ESCC patients prior of any treatment and 54 healthy controls for comparison with clinicopathological data through statistical analysis with Man Whitney U and Log-Rank tests. In addition, clinical value of MMP13 levels for diagnosis was evaluated by receiver operating characteristic (ROC) test. Results: The serum level of MMP-13 in patients (>250 pg/ml) was significantly higher than in the control group (Conclusions: These findings indicate a potential clinical significance of serum MMP13 measurement for early detection and prognostic assessment in ESCC patients.     

食管鳞癌细胞自分泌白介素-32通过诱导上皮间质转化促进肿瘤恶性表型

目的:探讨食管鳞癌（esophageal squamous cell carcinoma,ESCC）细胞自分泌的白介素-32（interleukin-32,IL-32）对细胞恶性表型的影响及其可能机制。方法:分析ESCC组织和正常组织中IL-32表达量差异,检测不同ESCC细胞系中IL-32表达;在高表达细胞中利用siRNA敲低IL-32表达,在低表达细胞系中过表达IL-32;通过MTT、Transwell实验等检测IL-32对ESCC细胞增殖、侵袭、迁移等表型的影响;Western blot检测上皮间质转化相关基因的表达变化。结果:ESCC组织中IL-32的表达量显著高于正常组织,ESCC细胞系中IL-32的表达量显著高于食管上皮永生化细胞系Het-1A;敲低IL-32显著抑制ESCC细胞的增殖、迁移和侵袭能力,过表达IL-32促进ESCC细胞的恶性表型;Western blot结果显示IL-32促进ESCC细胞发生上皮间质转化。结论:ESCC细胞自分泌的IL-32可能通过促进ESCC细胞发生上皮间质转化从而发挥促癌作用,抑制IL-32可能成为治疗ESCC的一种方法。