Chronic hepatitis B virus (HBV) infection is a global health problem that can lead to liver dysfunction, including liver cirrhosis and hepatocellular carcinoma (HCC). Current antiviral therapies can control viral replication in patients with chronic HBV infection; however, there is a risk of HCC development. HBV-related proteins may be produced in hepatocytes regardless of antiviral therapies and influence intracellular metabolism and signaling pathways, resulting in liver carcinogenesis. To understand the mechanisms of liver carcinogenesis, the effect of HBV infection in human hepatocytes should be analyzed. HBV infects human hepatocytes through transfer to the sodium taurocholate co-transporting polypeptide (NTCP). Although the NTCP is expressed on the hepatocyte surface in several animals, including mice, HBV infection is limited to human primates. Due to this species-specific liver tropism, suitable animal models for analyzing HBV replication and developing antivirals have been lacking since the discovery of the virus. Recently, a humanized mouse model carrying human hepatocytes in the liver was developed based on several immunodeficient mice; this is useful for analyzing the HBV life cycle, antiviral effects of existing/novel antivirals, and intracellular signaling pathways under HBV infection. Herein, the usefulness of human hepatocyte chimeric mouse models in the analysis of HBV-associated hepatocarcinogenesis is discussed.

• HBV
• HBs
• HBx
• HCC
• humanized mouse

• Animals
• Carcinogenesis/metabolism
• Carcinoma, Hepatocellular/metabolism
• Carcinoma, Hepatocellular/virology
• Disease Models, Animal
• Genotype
• Hepatitis B/metabolism
• Hepatitis B/virology
• Hepatitis B virus
• Hepatitis B, Chronic/metabolism
• Hepatitis B, Chronic/virology
• Hepatocytes/virology
• Humans
• Liver
• Liver Cirrhosis/virology
• Liver Neoplasms/metabolism
• Liver Neoplasms/virology
• Mice
• Symporters
• Taurocholic Acid
• Virus Replication

• Cancer
• Immunotherapeutics
• Inflammatory modulators
• NF-κβ
• Thymoquinone
• uncontrolled proliferation

• Prohibitin
• cholangiocarcinoma
• cholestatic liver injury
• hepatocellular carcinoma

Liver cancer (LC), which is one of the most common types of cancer worldwide, is notorious for its high morbidity and mortality rates. Interleukin-8 (IL-8), an important member of the CXC chemokine family that was originally classified as a potent neutrophil chemoattractant, has been shown to serve an important role in inflammation, tumor growth, invasion and metastasis through interactions with its receptors. However, the expression and functional roles of IL-8 and its receptors, CXC chemokine receptor (CXCR) 1 and CXCR2 in the progression of liver cancer remain to be fully elucidated. In the present study, it was shown that the mRNA levels of IL-8, CXCR1 and CXCR2 were increased in peripheral blood mononuclear cells from patients with liver cancer compared with those from patients with cirrhosis or normal controls (P<0.05). Higher levels of CXCR1, CXCR2 and IL-8 were associated with advanced tumor stage and increased risk of lymph node or distant metastasis. Immunohistochemistry showed that the IL-8, CXCR1 and CXCR2 proteins were expressed in the cytoplasm of hepatoma cells at higher intensities than those of normal controls (P<0.05). The semi-quantitative analysis revealed that the relative mean density of hepatic IL-8, CXCR1 and CXCR2 staining in liver cancer was significantly increased compared with that in normal liver tissues (P<0.05). The analysis revealed that the mRNA expression of IL-8 was positively associated with that of CXCR1 (r=0.618; P<0.05) and CXCR2 (r=0.569; P<0.05). The mRNA levels of CXCR1 and CXCR2 gradually increased with elevated expression of IL-8 in liver cancer. Experiments were performed using human Huh-7 and HepG2 cell lines, incubating cells with IL-8 and conducting in vitro migration and invasion assays. The results showed that the wound healing activity and migration of Huh-7 and HepG2 cells were increased by IL-8. Pretreatment of the cells with anti-CXCR1 or anti-CXCR2 (5 µM) for 30 min markedly inhibited IL-8-directed cell migration. Taken together, these results indicated that IL-8 promotes liver cancer cell migration via CXCR1 and CXCR2 and that targeting the CXCR1/2 may be a potential strategy for liver cancer treatment.

• CXC chemokine receptor 1
• CXC chemokine receptor 2
• interleukin-8
• invasion
• liver cancer
• metastasis

Epithelial-to-mesenchymal transition (EMT) is a complex process involving multiple genes, steps and stages. It refers to the disruption of tight intercellular junctions among epithelial cells under specific conditions, resulting in loss of the original polarity, order and consistency of the cells. Following EMT, the cells show interstitial cell characteristics with the capacity for adhesion and migration, while apoptosis is inhibited. This process is critically involved in embryogenesis, wound-healing, tumor invasion and metastasis. The tumor microenvironment is composed of infiltrating inflammatory cells, stromal cells and the active medium secreted by interstitial cells. Most patients with hepatocellular carcinoma (HCC) have a history of hepatitis virus infection. In such cases, major components of the tumor microenvironment include inflammatory cells, inflammatory factors and virus-encoded protein are major components. Here, we review the relationship between EMT and the inflammatory tumor microenvironment in the context of HCC. We also further elaborate the significant influence of infiltrating inflammatory cells and inflammatory mediators as well as the products expressed by the infecting virus in the tumor microenvironment on the EMT process.

• Epithelial-to-mesenchymal transition
• Expressed products of infected virus
• Hepatocellular carcinoma
• Inflammatory microenvironment
• Tumor-related macrophages

Hypoxia plays a critical role in many cancers. Hypoxia inducible factor-1α (HIF-1α) is an important mediator of the hypoxia response. It regulates the expression of various chemokines involved in tumor growth, angiogenesis and metastasis but the associated pathway needs further investigation.

The expression level of HIF-1α was determined in hepatocellular carcinoma (HCC) cells. The correlation of interleukin-8 (IL-8) and HIF-1α was assessed by knocking down HIF-1α. These cells were also used to assess its influence on HCC cell migration and invasion was checked. Pyrrolidinedithiocarbamate (PDTC), an inhibitor of NF-κB, was used to confirm the associated signaling pathway.

HIF-1α was significantly expressed in HCC cells and found to promote HCC cell migration and invasion in an IL-8-dependent manner. NF-κB was confirmed to be involved in the process.

HIF-1α promotes HCC cell migration and invasion by modulating IL-8 via the NF-κB pathway.

• HIF-1α
• Hepatocellular carcinoma
• Hypoxia
• IL-8
• NF-κB

The purpose of the present study was to investigate the role of latency-associated peptide (LAP)⁺CD4⁺T cells in hepatocellular carcinoma (HCC) immunity. Flow cytometric analysis was performed to detect the proportion of LAP⁺CD4⁺ T cells among the peripheral blood mononuclear cells (PBMCs) of 30 HBV-infected HCC patients at the pre-operative and post-operative stages, as well as 30 hepatitis B virus (HBV)-infected volunteers as a control group. Furthermore, tumor tissues and peri-tumor tissues from 28 patients with HCC, as well as hepatic tissues from 28 HBV-infected patients with benign lesions were subjected to immunohistochemical analysis with double staining for LAP and CD4, and the average number of the LAP⁺CD4⁺T cells in each visual field was quantified. The results indicated that the proportion of LAP+CD4+ T cells in the PBMCs of patients with HCC was significantly higher than that in the control group (1.84±0.85 vs. 0.73±0.39%, P=0.019), while it was significantly reduced after the operation (1.07±0.35, P=0.021), but still slightly, if not significantly, higher compared with that in the control group (P=0.342). Furthermore, the number of LAP⁺CD4⁺ T cells per high-magnification microscopic field (magnification, ×400) in the HCC tissues was 11.25±3.00, which was significantly higher than that in the peri-cancer tissues (5.75±1.00) and that in the HBV-infected hepatic tissues around benign lesions (2.61±0.83). In peri-cancer tissues, LAP⁺CD4⁺ T cells were also significantly more abundant than in control tissues. Furthermore, in the HCC tissues, LAP⁺CD4⁺ T cells were present as clusters in the tumor stroma and closely associated with CD4⁺ T lymphocytes. By contrast, in the peri-cancer liver tissues and HBV-infected hepatic tissues around benign lesions, LAP⁺CD4⁺ T cells were sparsely distributed. LAP⁺CD4⁺ T cells have marked inhibitory effects, and in the peripheral blood and tumor tissues of patients with HCC, they have an important role in the suppression of anti-tumor immunity and in the immune evasion of tumor cells.

• LAP+CD4+ T cells
• flow cytometry
• hepatocellular carcinoma
• immunohistochemistry
• peripheral blood mononuclear cells
• tumor microenvironment

Tumor metastasis is a multistep process involving a number of genetic alterations so that the genetic diagnosis is got increasingly attentions today. The aim of this study was to use RNA-seq to screen the effective differential expression genes in the peripheral blood mononuclear cells for the hepatic carcinoma with metastasis. The results showed that hepatic carcinoma samples gathered according to different metastasis. CCL3, CCL3L1, JUN, IL8, and IL1B were identified in inflammation mediated by chemokine and cytokine signaling pathway (P00031) in the hepatic carcinoma samples with metastasis, and subsequently confirmed by quantitative real-time polymerase chain reaction. In conclusions, CCL3, CCL3L1, JUN, IL8, and IL1B have the potential to be considered as candidates for future molecular diagnosis of the hepatic carcinoma with metastasis. This work may provide us with new visions into the metastasis process and potential efficient clinical diagnosis in the future.

• PBMC
• diagnosis
• hepatic carcinoma
• metastasis
• tumor heterogeneity

Hypoxia inducible factor-1α (HIF-1α), induces cytokines such as CXCL8 and tumor dissemination, chemo- and radio-resistance. We analyzed correlation between HIF-1α and CXCL8 levels, tumor characteristics and overall survival in 102 hepatocellular carcinoma (HCC) patients. Levels of HIF-1α and CXCL8 were increased in HCC tissues and cell lines. Patients with high levels of HIF-1α and CXCL8 had worse outcome and poorer prognosis than those with lower levels. Co-overexpression of HIF-1α and CXCL8 was an independent negative prognostic factor for overall and disease-free survival. HIF-1α silencing and CXCL8 siRNA decreased migration under hypoxic conditions in vitro. Hypoxia-induced activation of AKT/mTOR/STAT3 pathways was reversed by depletion of CXCL8. We conclude that HIF-1α and CXCL8 induce HCC progression and metastasis, associated with activation of AKT/mTOR/STAT3. Co-expression of HIF-1α and CXCL8 is a prognostic marker and a potential therapeutic target in HCC.

• HCC
• IL-8
• cytokines
• hypoxia
• metastasis
• migration

• Angiogenesis
• Cell migration
• HCC
• HCC xenograft
• HSCs
• Hepatic stellate cells
• Hepatocellular carcinoma
• LX-2
• Met
• PAR2
• Proteinase-activated receptor 2
• Receptor tyrosine kinase
• Src

Hepatocellular carcinoma (HCC) is a common malignancy and now the second commonest global cause of cancer death. HCC tumorigenesis is relatively silent and patients experience late symptomatic presentation. As the option for curative treatments is limited to early stage cancers, diagnosis in non-symptomatic individuals is crucial. International guidelines advise regular surveillance of high-risk populations but the current tools lack sufficient sensitivity for early stage tumors on the background of a cirrhotic nodular liver. A number of novel biomarkers have now been suggested in the literature, which may reinforce the current surveillance methods. In addition, recent metabonomic and proteomic discoveries have established specific metabolite expressions in HCC, according to Warburg’s phenomenon of altered energy metabolism. With clinical validation, a simple and non-invasive test from the serum or urine may be performed to diagnose HCC, particularly benefiting low resource regions where the burden of HCC is highest.

• Hepatocellular carcinoma
• Biomarker
• Metabonomics
• Warburg hypothesis
• Serum
• Plasma
• Urine

• MC_PC_12015 Medical Research Council
• MC_UP_A900_1120 Medical Research Council

• angiogenesis
• gastric cancer
• interleukin-8
• vascular endothelial growth factor receptor-1
• vascular endothelial growth factor receptor-2
• vascular endothelial growth factor-A

Most chronic liver diseases (CLDs) are characterized by inflammatory processes with aberrant expressions of various pro- and anti-inflammatory mediators in the liver. These mediators are the driving force of many inflammatory liver disorders, which often result in fibrosis, cirrhosis, and liver tumorigenesis. c-Myc is involved in many cellular events such as cell growth, proliferation, and differentiation. c-Myc upregulates IL-8, IL-10, TNF-α, and TGF-β, while IL-1, IL-2, IL-4, TNF-α, and TGF-β promote c-Myc expression. Their interactions play a central role in fibrosis, cirrhosis, and liver cancer. Molecular interference of their interactions offers possible therapeutic potential for CLDs. In this review, current knowledge of the molecular interactions between c-Myc and various well known inflammatory mediators is discussed.

• Cancer
• GPCR
• HepG2
• IL-8
• liver

• Cell Line
• Cyclooxygenase 2/metabolism
• Humans
• Interleukin-8/metabolism
• MAP Kinase Signaling System
• Up-Regulation
• Viral Nonstructural Proteins/physiology

• 3D culture
• Angiogenesis
• Cancer microenvironment
• Hypoxia
• Inflammation
• Tissue engineering

• Alginates/chemistry
• Biocompatible Materials/chemistry
• Cell Culture Techniques
• Cell Hypoxia
• Endothelium, Vascular/pathology
• Gene Expression Profiling
• Gene Expression Regulation, Neoplastic
• Glucuronic Acid/chemistry
• Hexuronic Acids/chemistry
• Human Umbilical Vein Endothelial Cells
• Humans
• Hydrogels/chemistry
• Inflammation/pathology
• Interleukin-8/metabolism
• Neoplasm Invasiveness
• Neoplasms/pathology
• Neovascularization, Pathologic
• Oligonucleotide Array Sequence Analysis
• Oxygen/chemistry
• Signal Transduction
• Tissue Engineering/methods
• Tumor Cells, Cultured

• R21 CA157383 NCI NIH HHS
• 1R01 CA185293 NCI NIH HHS
• U54CA143876 NCI NIH HHS
• R01 CA185293 NCI NIH HHS
• RC1 CA146065 NCI NIH HHS
• R21CA161532 NCI NIH HHS
• RC1 CA 146065 NCI NIH HHS
• U54 CA143876 NCI NIH HHS
• R21 CA161532 NCI NIH HHS
• R01 CA173083 NCI NIH HHS

Inflammatory factors play a vital role in the progression of liver cancer, although exact factors and related mechanisms still remain unclear. The present study aimed at screening inflammatory factors related to liver cancer metastasis and investigating the potential mechanism by which cancer cells are recruited. We screened and validated inflammatory factors by microarray and RT-PCR. Small interfering RNA (siRNA) and recombinant protein were used to assess CXCL5 effects on the movement of liver cancer cells (LCCs). Our screening microarray demonstrated over-expression of CXCL5 in LCCs with high metastatic potentials. CXCL5 increased LCCs migration and invasion, probably through autocrine and paracrine mechanisms. CXCL5-CXCR2 and ERK1/2 pathways could play critical roles in the regulation of LCCs migration. Our data indicates that LCCs per se may act as the producer and receptor of CXCL5 responsible for liver cancer migration and invasion.

The biological role of interleukin-37 (IL-37) in cancer is large unknown. Through immunohistochemical detection using 163 primary hepatocellular carcinoma (HCC) clinical specimens, we found the expression of IL-37 was decreased in tumor tissues, and the expression level was negatively correlated with tumor size. High expression of IL-37 in HCC tumor tissues was associated with better overall survival (OS) and disease-free survival (DFS). IL-37 expression in tumor tissues was positively associated with the density of tumor-infiltrating CD57+ natural killer (NK) cells, but not with the CD3+ and CD8+ T cells. Consistently, in vitro chemotaxis analysis showed that IL-37- overexpressing HCC cells could recruit more NK cells. The in vivo mouse model experiments also revealed that overexpression IL-37 in HCC cells significantly delayed tumor growth and recruited more NK cells into tumors tissues. Our finding suggested that IL-37 might play an important role for the prognosis of HCC patients via regulating innate immune-action.

Hepatocellular carcinoma is the fifth most common cancer worldwide and shows a complex clinical course, poor response to pharmacological treatment, and a severe prognosis. Thus, the aim of this study was to investigate whether tacrolimus (FK506) has synergistic antitumor effects with doxorubicin on two human hepatocellular carcinoma cell lines, Huh7 and HepG2. Cell viability was analyzed by Sulforhodamine B assay and synergic effect was evaluated by the software CalcuSyn. Cell apoptosis was evaluated using Annexin V and Dead Cell assay. Apoptosis-related protein PARP-1 cleaved and autophagy-related protein expressions (Beclin-1 and LC3B) were measured by western blotting analysis. Cytokines concentration in cellular supernatants after treatments was studied by Bio-Plex assay. Interestingly the formulation with doxorubicin and tacrolimus induced higher cytotoxicity level on tumor cells than single treatment. Moreover, our results showed that the mechanisms involved were (i) a strong cell apoptosis induction, (ii) contemporaneous decrease of autophagy activation, understood as prosurvival process, and (iii) downregulation of proinflammatory cytokines. In conclusion, future studies could relate to the doxorubicin/tacrolimus combination effects in mice models bearing HCC in order to see if this formulation could be useful in HCC treatment.

• Apoptosis/drug effects
• Benzoquinones/pharmacology
• Carcinoma, Hepatocellular/drug therapy
• Carcinoma, Hepatocellular/metabolism
• Caspase 3/metabolism
• Caspase 9/metabolism
• Cell Line, Tumor
• Cell Proliferation/drug effects
• G2 Phase Cell Cycle Checkpoints/drug effects
• Hep G2 Cells
• Humans
• Interleukin-8/metabolism
• Liver Neoplasms/drug therapy
• Liver Neoplasms/metabolism
• M Phase Cell Cycle Checkpoints/drug effects
• NF-kappa B/metabolism
• Oxidative Stress/drug effects
• Poly(ADP-ribose) Polymerases/metabolism
• Proto-Oncogene Proteins c-bcl-2/metabolism
• Reactive Oxygen Species/metabolism
• Receptors, TNF-Related Apoptosis-Inducing Ligand/metabolism
• bcl-X Protein/metabolism

• Adult
• Aged
• Alleles
• Carcinoma, Hepatocellular/genetics
• Carcinoma, Hepatocellular/physiopathology
• China
• Female
• Genetic Association Studies
• Genetic Predisposition to Disease
• Haplotypes
• Humans
• Interleukin-8/genetics
• Liver Cirrhosis/genetics
• Liver Neoplasms/genetics
• Liver Neoplasms/physiopathology
• Male
• Middle Aged
• Polymorphism, Single Nucleotide

Hepatocellular carcinoma (HCC) related to hepatitis B virus (HBV) and hepatitis C virus (HCV) infections is thought to account for more than 80% of primary liver cancers. Both HBV and HCV can establish chronic liver inflammatory infections, altering hepatocyte and liver physiology with potential liver disease progression and HCC development. Cyclophilin A (CypA) has been identified as an essential host factor for the HCV replication by physically interacting with the HCV non structural protein NS5A that in turn interacts with RNA-dependent RNA polymerase NS5B. CypA, a cytosolic binding protein of the immunosuppressive drug cyclosporine A, is overexpressed in many cancer types and often associated with malignant transformation. Therefore, CypA can be a good target for molecular cancer therapy. Because of antiviral activity, the CypA inhibitors have been tested for the treatment of chronic hepatitis C. Nonimmunosuppressive Cyp inhibitors such as NIM811, SCY-635, and Alisporivir have attracted more interests for appropriating CypA for antiviral chemotherapeutic target on HCV infection. This review describes CypA inhibitors as a potential HCC treatment tool that is contrived by their obstructing chronic HCV infection and summarizes roles of CypA in cancer development.

• Cyclophilin A
• Hepatitis C virus
• Hepatocellular carcinoma
• Peptidyl prolyl isomerase

• selenite
• selenbp1
• selk
• gpx1
• hepg2
• huh7
• hepatocarcinoma
• cytokines
• interactomics
• inflammation

• Carcinoma, Hepatocellular/metabolism
• Cell Line, Tumor
• Cytokines/metabolism
• Glutathione Peroxidase/metabolism
• Hep G2 Cells
• Hepatocytes/drug effects
• Hepatocytes/metabolism
• Humans
• Liver Neoplasms/metabolism
• Protein Binding
• Protein Interaction Mapping
• Protein Interaction Maps
• Selenium-Binding Proteins/metabolism
• Selenoproteins/metabolism
• Sodium Selenite/pharmacology
• Glutathione Peroxidase GPX1

Interleukin (IL)-8 plays the critical role in the initiation of micro-environmental inflammation responsible for tumour growth and patient prognosis. This study aimed at investigating the molecular mechanisms of IL-8 production from human hepatocellular carcinoma (HCC) cells. The levels of IL-8 and phosphorylation of p38 mitogen-activated protein kinase (MAPK), ERK1/2 and Akt in MHCC-97H cells were measured by ELISA, Western blot and immunofluorescence. NF-κB p65 protein nuclear translocation was determined by non-radioactive NF-κB p50/p65 transcription factor activity kit and cell bio-behaviours were detected by the real-time cell-monitoring system. Tumour necrosis factor-α (TNF-α) significantly induced phosphorylation of p38 MAPK, ERK, Akt and production of IL-8 from HCC cells, which were prevented by SB203580 (p38 MAPK inhibitor), PD98059 (ERK inhibitor), LY294002 and Wortmannin (PI3K inhibitor) and SB328437 (CCR3 inhibitor). TNF-α could significantly increase the translocation of NF-κB p65 protein into the nucleus in a dose-dependent manner, while SB203580 partially inhibited. In inflammatory micro-environment, HCC auto-produced IL-8 through p38 MAPK, ERK and PI3K/Akt signalling pathways, where the p38 MAPK is a central factor to activate the NF-κB pathway and regulate the expression of IL-8 production. There was a potential cross-talking between receptors.

• Aged
• C-Reactive Protein/metabolism
• Carcinoma, Hepatocellular/blood
• Carcinoma, Hepatocellular/complications
• Carcinoma, Hepatocellular/diagnosis
• Diagnostic Errors
• Fever/etiology
• Humans
• Liver Abscess/diagnosis
• Liver Neoplasms/blood
• Liver Neoplasms/complications
• Liver Neoplasms/diagnosis
• Male
• Sarcoma/blood
• Sarcoma/complications
• Sarcoma/diagnosis

Hepatocellular carcinoma (HCC) is among the most aggressive and fatal cancers. Its treatment with conventional chemotherapeutic agents is inefficient, due to several side effects linked to impaired organ function typical of liver diseases. Consequently, there exists a decisive requirement to explore possible alternative chemopreventive and therapeutic strategies. The use of dietary antioxidants and micronutrients has been proposed for HCC successful management. The aim of this work was to test in vitro the effects of lipoic acid, caffeic acid and a new synthesized lipoyl-caffeic conjugate on human hepatoma cell lines in order to assess their effect on tumor cell growth. The results of cytotoxicity assays at different times showed that the cell viability was directly proportional to the molecule concentrations and incubation times. Moreover, to evaluate the pro- or anti-inflammatory effects of these molecules, the cytokine concentrations were evaluated in treated and untreated cellular supernatants. The obtained cytokine pattern showed that, at the increasing of three molecules concentrations, three pro-inflammatory cytokines such as IL-1β, IL-8 and TNF-α decreased whereas the anti-inflammatory cytokine such as IL-10 increased.

Serum interleukin-8 (IL-8) and interferon-alpha (IFN-α) levels have been estimated from a total of 88 individuals of which 19 were disease-free healthy individuals, and 69 were patients with thyroid diseases: goitre (N = 21), autoimmune diseases (N = 16), and carcinomas (N = 32). Both IL-8 and IFN-α were significantly higher in all the patients as compared to healthy individuals. Serum IL-8 levels showed significant positive correlation with disease stage in thyroid cancer patients. Higher serum IL-8 levels were associated with advanced disease stage while no significant correlation was observed between serum IFN-α levels and any of the clinicopathological parameters. IL-8 and IFN-α significantly correlated with each other in anaplastic carcinoma patients. Finally concluding, monitoring the serum IL-8 and IFN-α levels can help differentiate patients with thyroid diseases from healthy individuals, and IL-8 seems to have a role in the pathogenesis of thyroid diseases and may represent a target for innovative diagnostic and therapeutic strategies.

We discuss a patient who had poorly differentiated HCC with pyrexia and high CRP in laboratory data, which are not commonly observed in the usual HCC. A 50-year-old man with a history of liver dysfunction was admitted with a chief complaint of a prolonged fever and general fatigue. Preoperative diagnosis was HCC with portal vein tumor thrombus. Posterior segmentectomy of the liver and thrombectomy was performed. Rapid tumor recurrence occurred after surgery, and he died 79 days after the operation. Immunohistochemical stain of HCC in this patient revealed the production of proinflammatory cytokine, interleukin-8 (IL-8). IL-8 production may have contributed to the high fever, high inflammatory reaction, and poor prognosis in this case.

AIM: To study the implication of prokineticin 1 (PK1/EG-VEGF) and prokineticin 2 (PK2/Bv8) in hepatocellular carcinoma angiogenesis.

METHODS: The gene induction of PK1/EG-VEGF and PK2/Bv8 was investigated in 10 normal, 28 fibrotic and 28 tumoral livers by using real time PCR. Their expression was compared to the expression of VEGF (an angiogenesis marker), vWF (an endothelial cell marker) and to CD68 (a monocyte/macrophage marker). Furthermore, the mRNA levels of PK1/EG-VEGF, PK2/Bv8, prokineticin receptor 1 and 2 were evaluated by real time PCR in isolated liver cell populations. Finally, PK2/Bv8 protein was detected in normal liver paraffin sections and in isolated liver cells by immunohistochemistry and immunocytochemistry.

RESULTS: PK2/Bv8 mRNA but not PK1/EG-VEGF was expressed in all types of normal liver samples examined. In the context of liver tumor development, we reported that PK2/Bv8 correlates only with CD68 and showed a significant decrease in expression as the pathology evolves towards cancer. Whereas, VEGF and vWF mRNA were significantly upregulated in both fibrosis and HCC, as expected. In addition, out of all isolated liver cells examined, only Kupffer cells (liver resident macrophages) express significant levels of PK2/Bv8 and its receptors, prokineticin receptor 1 and 2.

CONCLUSION: In normal liver PK2/Bv8 and its receptors were specifically expressed by Kupffer cells. PK2/Bv8 expression decreased as the liver evolves towards cancer and did not correlate with HCC angiogenesis.

• Prokineticin
• Hepatocellular carcinoma
• PK2/Bv8
• Angiogenesis
• Kupffer cells
• Vascular endothelial growth factor
• Liver

AIM: To determine the expression of macro-phage migration inhibitory factor (MIF) in hepatocellular carcinoma (HCC) and its effect on cell proliferation and angiogenesis.

METHODS: The protein expression of MIF, interleukin-8 (IL-8), and vascular endothelial factor (VEGF) as well as microvessel density (MVD) were examined by immunochemistry in HCC tissues (n = 40), cancer-adjacent liver tissues (n = 40) and normal liver tissues (n = 10). At the same time, real time quantitative polymerase chain reaction (RQ-PCR) was used to detect the mRNA levels of MIF, VEGF and IL-8. After human hepatocarcinama cell line Bel-7402 was treated with recombinant MIF (rMIF), cell proliferation was detected by MTT assay and the secretion levels and mRNA expression of VEGF and IL-8 were examined by enzyme linked immunosorbent assay (ELISA) and RQ-PCR, respectively.

RESULTS: The positive rates of MIF, IL-8 and VEGF in HCC tissues were significantly higher than those in normal liver tissues (72.5% vs 30.0%, 67.5% vs 20.0%, 62.5% vs 30.0%, all P < 0.01). The expression of MIF and IL-8 were markedly correlated with VEGF expression (r = 0.271, P < 0.05) and MVD (r = 0.284, P < 0.05). VEGF concentration in the supernatant of Bel-7402 cells was higher (900.00 ± 264.58 ng/L) than that in the controls (180 ± 50 ng/L) at the 24^th hour after rMIF treatment (P < 0.01). However, there was no significant variation in IL-8 concentration (P > 0.05). The post-treatment level of VEGF mRNA was 6.86 times as high as that without treatment (P < 0.05), and there was no significant change in IL-8 mRNA expression (P > 0.05).

CONCLUSION: The over-expression of MIF plays a crucial role in the angiogenesis of liver cancer and the roles of MIF and IL-8 may be realized via VEGF.

• Liver cancer
• Macrophage migration inhibitory factor
• Vascular endothelial factor
• Interleukin-8
• Microvessel density
• Immunochemistry
• Real time quantitative polymerase chain reaction

• Animals
• Antineoplastic Agents/pharmacology
• Cell Growth Processes/drug effects
• Chemokine CXCL1
• Chemokines, CXC/biosynthesis
• ErbB Receptors/metabolism
• Extracellular Signal-Regulated MAP Kinases/metabolism
• Gefitinib
• Gene Dosage
• Genes, erbB-1
• Intercellular Signaling Peptides and Proteins/biosynthesis
• Liver Neoplasms, Experimental/blood supply
• Liver Neoplasms, Experimental/drug therapy
• Liver Neoplasms, Experimental/metabolism
• Male
• Mice
• Mice, Inbred BALB C
• Neovascularization, Pathologic/drug therapy
• Neovascularization, Pathologic/metabolism
• Neovascularization, Pathologic/pathology
• PTEN Phosphohydrolase/metabolism
• Phosphorylation/drug effects
• Proto-Oncogene Proteins c-akt/metabolism
• Quinazolines/pharmacology
• Signal Transduction/drug effects
• Vascular Endothelial Growth Factor A/biosynthesis

• Angiogenesis Inhibitors/pharmacology
• Angiogenesis Inhibitors/therapeutic use
• Carcinoma, Hepatocellular/blood supply
• Carcinoma, Hepatocellular/drug therapy
• Carcinoma, Hepatocellular/pathology
• Humans
• Liver Neoplasms/blood supply
• Liver Neoplasms/drug therapy
• Liver Neoplasms/pathology
• Neovascularization, Pathologic/metabolism
• Neovascularization, Pathologic/prevention & control
• Vascular Endothelial Growth Factor A/antagonists & inhibitors

Hepatocellular carcinoma (HCC) is one of the most frequent malignant tumors and is the second most common cause of cancer death in China. Therefore, it is very important to detect this disease and the recurrence at its earlier period. Serum tumor markers, as the effective method for detecting hepatocellular carcinoma for a long time, could be divided into 4 categories: oncofetal antigens and glycoprotein antigens; enzymes and isoenzymes; genes; and cytokines. Serum alpha fetoprotein (AFP) is the most widely used tumor marker in detecting patients with hepatocellular carcinoma, and has been proven to have capability of prefiguring the prognosis. However, it has been indicated that AFP-L3 and DCP excel AFP in differentiating hepatocellular carcinoma from nonmalignant hepatopathy and detecting small hepatocellular carcinoma. Some tumor markers, such as human cervical cancer oncogene and human telomerase reverse transcriptase mRNA, have also been indicated to have higher accuracies than AFP. Furthermore, some other tumor markers, such as glypican-3, gamma-glutamyl transferase II, alpha-l-fucosidase, transforming growth factor-beta1, tumor-specific growth factor, have been indicated to be available supplementaries to AFP in the detection. AFP mRNA has been shown to correlate with the metastasis and recurrence of HCC, and it may be the most useful marker to prefigure the prognosis. Some other markers, such as gamma-glutamyl transferase mRNA, vascular endothelial growth factor, and interleukin-8, could also be used as available prognostic indicators, and the simultaneous determination of AFP and these markers may detect the recurrence of HCC at its earlier period.

• Hepatocellular carcinoma
• Serum tumor markers
• Sensitivity
• Specificity
• Prognosis

• Antigens, Neoplasm/blood
• Biomarkers/blood
• Biomarkers, Tumor/blood
• Carcinoma, Hepatocellular/blood
• Carcinoma, Hepatocellular/diagnosis
• Cytokines/blood
• Glycoproteins/blood
• Humans
• Liver Neoplasms/blood
• Liver Neoplasms/diagnosis
• Prognosis
• Protein Precursors/blood
• Prothrombin
• RNA, Messenger/blood
• alpha-Fetoproteins/analysis
• alpha-Fetoproteins/genetics
• gamma-Glutamyltransferase/blood
• gamma-Glutamyltransferase/genetics

• Aflatoxin B1/toxicity
• Animals
• Carcinoma, Hepatocellular/chemically induced
• Carcinoma, Hepatocellular/genetics
• Carcinoma, Hepatocellular/pathology
• Disease Models, Animal
• Gene Expression Profiling
• Gene Expression Regulation, Neoplastic/drug effects
• Liver/drug effects
• Liver/pathology
• Liver Neoplasms/chemically induced
• Liver Neoplasms/genetics
• Liver Neoplasms/pathology
• Oligonucleotide Array Sequence Analysis
• Oncorhynchus mykiss/genetics
• RNA, Messenger/metabolism
• RNA, Neoplasm/analysis
• Reverse Transcriptase Polymerase Chain Reaction
• Transcription, Genetic/drug effects

• CA90890 NCI NIH HHS
• ES00210 NIEHS NIH HHS
• ES03850 NIEHS NIH HHS
• ES07060 NIEHS NIH HHS
• ES11267 NIEHS NIH HHS

• Adenocarcinoma, Clear Cell/drug therapy
• Animals
• Antiviral Agents/therapeutic use
• Apoptosis
• Cell Cycle
• Cell Line, Tumor
• Cell Proliferation
• Dose-Response Relationship, Drug
• Enzyme-Linked Immunosorbent Assay
• Female
• Fibroblast Growth Factor 2/metabolism
• Flow Cytometry
• Humans
• Interferon Type I/therapeutic use
• Interferon-alpha
• Interleukin-8/metabolism
• Membrane Proteins
• Mice
• Mice, Inbred BALB C
• Mice, Nude
• Microcirculation
• Necrosis
• Ovarian Neoplasms/drug therapy
• Ovarian Neoplasms/metabolism
• RNA, Messenger/metabolism
• Receptor, Interferon alpha-beta
• Receptors, Interferon
• Recombinant Proteins
• Reverse Transcriptase Polymerase Chain Reaction
• Time Factors
• Vascular Endothelial Growth Factor A/metabolism

AIM: To investigate the relationship among the expression of IL-8, MCP-1, MIP-1a and tumor-associated macrophage (TAM) counting in pancreatic cancer.

METHODS: Paraffin section of pancreatic cancer specimen was made using routine method. TAM was counted after staining. The expression of IL-8, MCP-1, MIP-1a was detected by immunohistochemical method

RESULTS: The positive rates of IL-8, MCP-1 and MIP-1a were 62.7%, 66.7% and 66.7%, and their scores were 2.9±1.9, 2.6±1.8 and 2.4±1.7, respectively. Mean counting of TAM was 18.0±6.0 in each pancreatic cancer specimen. The scores of IL-8 and TAM counting were significantly less in well-differentiated cancer than those in poorly-differentiated cancer (IL-8:2.7±1.7 vs 3.2±1.2, P <0.05; TAM:15.9±6.4 vs 21.2±5.2, P <0.05). The positive rates and levels of IL-8, MCP-1, MIP-1a expression as well as TAM counting were significantly higher in metastatic cancer than those in the cancer without metastasis (P <0.05 or P <0.01). TAM counting was significantly higher in IL-8, MCP-1 and MIP-1a positive cancer than that in the negative one. The levels of IL-8, MCP-1, MIP-1a expression positively related to TAM counting (r_IL-8 = 0.52, P <0.01; r_MCP-1 = 0.50, P <0.01; r_MIP-1a = 0.46, P <0.01). At the same time, positive relationship existed between expression of IL-8 and MCP-1a (r = 0.52, P <0.01), IL-8 and MCP-1 (r = 0.54, P <0.01), MCP-1 and MCP-1a (r = 0.64, P <0.01).

CONCLUSION: The expression of IL-8, MCP-1, MIP-1a and TAM counting may relate to the progression, angiogenesis, metastasis and prognosis of pancreatic cancer. Positive expression of IL-8, MCP-1, MIP-1a and high TAM counting may indicate rapid progression and poor prognosis of pancreatic cancer.

• N/A

AIM: To evaluate the diagnostic value of assaying telomerase activity in ascites cells for the differential diagnosis of malignant and non-malignant ascites.

METHODS: Ascites from 40 patients with hepatocellular carcinoma (HCC), 31 with non-HCC gastrointestinal carcinoma (CA), and 24 with liver cirrhosis (LC) were analyzed for telomerase activity. The telomerase activities in cell pellets from ascites were measured according to the Telomeric Repeat Amplification Protocol (TRAP) and quantified with a densitometer.

RESULTS: Positive telomerase activity was detected in 16 of 31 (52%) CA patients, 10 of 40 (25%) HCC patients, and 1 of 24 (4%) LC patients (P < 0.001). The telomerase activity was higher in the ascites of CA patients than in the ascites of HCC or LC patients (CA: 22.9 ± 5.8, HCC: 6.7 ± 2.5, LC: 1.3 ± 1.3, P = 0.001). Cytology was positive in 18 CA patients (58%) and 1 HCC patient (2.5%), respectively. The positive telomerase activity was not related to patients’ age, gender, and ascitic protein concentration, but to white blood count (r = 0.31, P = 0.002), neutrophil count (r = 0.29, P = 0.005), and the C-reactive protein level (r = 0.29, P = 0.018). When the results of both cytological examination and telomerase assay were considered together, the sensitivity increased to 77% for CA patients, 25% for HCC patients, and 48% for all 71 gastrointestinal cancer patients.

CONCLUSION: Combining cytological examination of ascites with telomerase activity assay significantly improves the differential diagnosis between malignant and non-malignant ascites.

• Aged
• Ascites/diagnosis
• Ascites/etiology
• Ascites/metabolism
• Ascitic Fluid/metabolism
• Carcinoma, Hepatocellular/complications
• Carcinoma, Hepatocellular/diagnosis
• Carcinoma, Hepatocellular/metabolism
• Diagnosis, Differential
• Female
• Gastrointestinal Neoplasms/complications
• Gastrointestinal Neoplasms/diagnosis
• Gastrointestinal Neoplasms/metabolism
• Humans
• Liver Cirrhosis/complications
• Liver Cirrhosis/diagnosis
• Liver Cirrhosis/metabolism
• Liver Neoplasms/complications
• Liver Neoplasms/diagnosis
• Liver Neoplasms/metabolism
• Male
• Middle Aged
• Prospective Studies
• Telomerase/metabolism

• Base Sequence
• Carcinoma, Hepatocellular/genetics
• Carcinoma, Hepatocellular/metabolism
• Carcinoma, Hepatocellular/pathology
• Chemokine CCL3
• Chemokine CCL4
• DNA Primers
• Humans
• Interleukin-1/physiology
• Liver Neoplasms/genetics
• Liver Neoplasms/metabolism
• Liver Neoplasms/pathology
• Macrophage Inflammatory Proteins/genetics
• Macrophage Inflammatory Proteins/metabolism
• Polymerase Chain Reaction
• RNA, Messenger/genetics
• Receptors, CCR1
• Receptors, Chemokine/genetics
• Receptors, Chemokine/metabolism
• Reverse Transcriptase Polymerase Chain Reaction
• Transcription, Genetic
• Tumor Cells, Cultured