agar and Liver-Neoplasms

CT-guided radiofrequency ablation (RFA) is a potentially curative minimally invasive treatment for liver cancer. Local tumor recurrence limits the success of RFA for large or irregular tumors as it is difficult to visualize the tissue destroyed. This study was designed to validate a real-time software-simulated ablation volume for intraprocedural guidance.. Software that simulated RFA physics calculated ablation volumes in 17 agar-albumin phantoms (7 with a simulated vessel) and in six in-vivo (porcine) ablations. The software-modeled volumes were compared with the actual ablations (physical lesion in agar, contrast CT in the porcine model) and to the volume predicted by the manufacturer's charts. Error was defined as the distance from evenly distributed points on the segmented true ablation volume surfaces to the closest points on the corresponding computer-generated model, and for the porcine model, to the manufacturer-specified ablation volume.. The average maximum error of the simulation was 2.8 mm (range to 4.9 mm) in the phantoms. The heat-sink effect from the simulated vessel was well-modeled by the simulation. In the porcine model, the average maximum error of the simulation was 5.2 mm (range to 8.1 mm) vs 7.8 mm (range to 10.0mm) for the manufacturer's model (p = 0.009).. A real-time computer-generated RFA model incorporated tine position, energy deposited, and large vessel proximity to predict the ablation volume in agar phantoms with less than 3mm maximum error. Although the in-vivo model had slightly higher maximum error, the software better predicted the achieved ablation volume compared to the manufacturer's ablation maps.

Topics: Agar; Animals; Catheter Ablation; Liver; Liver Neoplasms; Software; Swine

The aim of this study was to characterize the oncogenic function and mechanism of Cathepsin Z (CTSZ) at 20q13.3, a frequently amplified region in hepatocellular carcinoma (HCC). Real-time PCR were used to compare CTSZ expression between paired HCC tumor and non-tumor specimens. CTSZ gene was stably transfected into HCC line QGY-7703 cells and its role in tumorigenicity and cell motility was characterized by soft agar, wound-healing, transwell invasion and cell adhesion assay, and tumor xenograft mouse model. Western blot analysis was used to study expression of proteins associated with epithelial-mesenchymal transition (EMT).Upregulation of CTSZ was detected in 59/137 (43%) of primary HCCs, which was significantly associated with advanced clinical stage (P = 0.000). Functional study found that CTSZ could increase colony formation in soft agar and promote cell motility. Further study found that the metastatic effect of CTSZ was associated with its role in inducing epithelial-mesenchymal transition (EMT) by upregulating mesenchymal markers (fibronectin and vimentin) and downregulating epithelial markers (E-cadherin and α-catenin). In addition, CTSZ could also upregulate proteins associated with extracellular matrix remodeling such as MMP2, MMP3 and MMP9. Taken together, our data suggested that CTSZ was a candidate oncogene within the 20q13 amplicon and it played an important role in HCC metastasis.

Topics: Agar; alpha Catenin; Animals; Cadherins; Carcinoma, Hepatocellular; Cathepsin Z; Cell Adhesion; Cell Movement; Epithelial-Mesenchymal Transition; Fibronectins; Gene Expression Regulation, Neoplastic; Humans; Liver Neoplasms; Male; Matrix Metalloproteinases; Mice; Mice, SCID; Neoplasm Metastasis; Neoplasm Transplantation; Vimentin; Wound Healing

Hepatitis B x antigen (HBxAg) is a trans-activating protein that contributes to liver cancer, in part, by altering the expression of cellular genes. However, few natural effectors of HBxAg have been identified. Hence, HBxAg positive and negative HepG2 cells were prepared and analyzed by PCR select cDNA subtraction. The results identified elevated vascular endothelial growth factor receptor-3 short form splice variant (VEGFR-3(S)) expression in HBxAg positive compared to negative cells. Normally, VEGFR-3 activates Akt signaling in lymphatic endothelial cells, resulting in lymphangiogenesis. In contrast, the results here show that the expression of VEGFR-3(S) is up-regulated in >75% of HBxAg positive hepatocellular carcinoma (HCC) nodules. VEGFR-3(S) up-regulation correlates with the expression of HBxAg, is associated with decreased survival in tumor bearing patients, and when over-expressed in HepG2 cells, strongly stimulated cell growth in culture, in soft agar, and accelerated tumor formation in a ligand independent manner. VEGFR-3(S) siRNA partially blocked the ability of HBxAg to promote hepatocellular growth. In conclusion, HBxAg may short circuit VEGFR-3(S) signaling in liver cancer. Blocking VEGFR-3(S) signaling may be effective in preventing tumor development and/or prolonging survival in tumor bearing patients.

Topics: Agar; Animals; Carcinoma, Hepatocellular; Cell Division; Cell Line, Tumor; Cloning, Molecular; Gene Expression Regulation, Neoplastic; Gene Expression Regulation, Viral; Hepatitis B; Hepatitis B virus; Humans; Liver; Liver Neoplasms; Mice; Mice, Nude; Proliferating Cell Nuclear Antigen; Proto-Oncogene Proteins c-akt; RNA, Small Interfering; Signal Transduction; Trans-Activators; Up-Regulation; Vascular Endothelial Growth Factor C; Vascular Endothelial Growth Factor Receptor-3; Viral Regulatory and Accessory Proteins

Hepatocellular carcinoma (HCC) is one of the most common malignancies in Southeast Asia. Although inactivation of pRb2/p130 has been reported in a variety of human cancers, its function in HCC has not been established. In this study we report that loss of expression of pRb2/p130 was detected by immunohistochemistry and western blotting in 15.2% (7 of 46) HCCs examined. High levels of pRb2/p130 expression were found in 84.8% (39 of 46) HCCs studied. Western blot analysis revealed that HCC had 3.5-fold higher pRb2/p130 than adjacent benign liver (ABL) tissues. 71.7% (33 of 46) of HCCs examined exhibited both nuclear and cytoplasmic staining for pRb2/p130. Cytoplasmic staining was found in 93.5% (43 of 46) of ABL tissues. Overproduction of pRb2/p130 in HepG2 cells led to growth suppression, cell cycle arrest in G0/G1, altered cell morphology, inhibition of in vitro colony formation and reduction in tumourigenicity in SCID mice. This demonstration suggests a role of pRb2/p130 as a tumour suppressor protein in HCC and the loss of this protein may lead to the development or progression of HCC. Overexpression of pRb2/p130 in HCC was, therefore, suggested to be a programmed protective response of the organism to uncontrolled proliferation.

Topics: Agar; Animals; Blotting, Western; Carcinoma, Hepatocellular; Cell Cycle; Cell Division; Cell Line; Cytoplasm; Disease Progression; DNA, Complementary; Electrophoresis, Agar Gel; Flow Cytometry; G1 Phase; Humans; Immunohistochemistry; Liver Neoplasms; Mice; Mice, SCID; Neoplasm Transplantation; Neoplasms; Precipitin Tests; Protein Biosynthesis; Proteins; Resting Phase, Cell Cycle; Retinoblastoma-Like Protein p130; Reverse Transcriptase Polymerase Chain Reaction; Time Factors; Transfection; Tumor Cells, Cultured

The central role of T cell activation in hepatocellular injury has been well documented. In this article, we provide evidence suggesting that T cells may also play a protective role in liver disease by releasing interleukin-22 (IL-22), a recently identified T cell-derived cytokine whose biological significance is unclear. IL-22 messenger RNA and protein expression are significantly elevated in T cell-mediated hepatitis induced by concanavalin A (ConA) but are less extensively elevated in the carbon tetrachloride-induced liver injury model. Activated CD3(+) T cells are likely responsible for the production of IL-22 in the liver after injection of ConA. The IL-22 receptor is normally expressed at high levels by hepatocytes and further induced after ConA injection. IL-22 blockade with a neutralizing antibody reduces signal transducer and activator of transcription factor 3 (STAT3) activation and worsens liver injury in T cell-mediated hepatitis, whereas injection of recombinant IL-22 attenuates such injury. In vitro treatment with recombinant IL-22 or overexpression of IL-22 promotes cell growth and survival in human hepatocellular carcinoma HepG2 cells. Stable overexpression of IL-22 in HepG2 cells constitutively activates STAT3 and induces expression of a variety of antiapoptotic (e.g., Bcl-2, Bcl-xL, Mcl-1) and mitogenic (e.g., c-myc, cyclin D1, Rb2, CDK4) proteins. Blocking STAT3 activation abolishes the antiapoptotic and mitogenic actions of IL-22 in hepatic cells. In conclusion, the T cell-derived cytokine IL-22 is a survival factor for hepatocytes; this suggests that T cell activation may also prevent and repair liver injury by releasing hepatoprotective cytokine IL-22 in addition to its previously documented central role in hepatocellular injury.

Topics: Agar; Animals; Carcinoma, Hepatocellular; Cell Division; Cell Line, Tumor; Cell Survival; Chemical and Drug Induced Liver Injury; Concanavalin A; DNA-Binding Proteins; Hepatitis; Hepatocytes; Humans; Interleukin-22; Interleukins; Liver Neoplasms; Mice; Mice, Nude; Neoplasm Transplantation; Receptors, Interleukin; STAT3 Transcription Factor; T-Lymphocytes; Trans-Activators

We have shown previously that approximately 1 in 10,000 primary hepatocytes isolated from untreated rats undergo clonal growth in soft agar in vitro in response to the synergistic action of nafenopin, a peroxisome proliferator (PP) and epidermal growth factor (EGF), a naturally occurring liver growth regulator. Here, we demonstrate that prior treatment of the animals with the genotoxic hepatocarcinogen diethylnitrosamine (DEN) caused a dose-dependent increase in soft agar colony numbers formed in vitro. These data suggest that the colony assay may offer a method of detecting in vitro hepatocytes transformed in vivo by DEN. It is known that rats treated with DEN develop enzyme altered foci prior to the development of tumours. The majority of these foci express high levels of gamma-glutamyl transpeptidase (GGT). However, foci promoted by PPs do not show this increased enzyme activity. In the present study, the colonies we have generated in vitro mimicked this pattern since the majority (approximately 80%) of the spontaneous colonies expressed GGT whereas colonies promoted by the synergistic action of nafenopin and EGF were mainly (75%) GGT negative. The proportion of colonies positive for GGT were similar using either hepatocytes isolated from control or from DEN-initiated rats. Further studies are required to assess if the hepatocytes selected for clonal expansion by this EGF/nafenopin regime reflect the presumed pre-neoplastic cells induced by genotoxin in vivo and associated with an increased propensity to cancer.

Topics: Agar; Animals; Carcinogenicity Tests; Carcinoma, Hepatocellular; Cells, Cultured; Contact Inhibition; Diethylnitrosamine; Dose-Response Relationship, Drug; Drug Synergism; Epidermal Growth Factor; gamma-Glutamyltransferase; Gene Expression Regulation, Neoplastic; Liver Neoplasms; Male; Nafenopin; Rats; Rats, Wistar; Tumor Stem Cell Assay

To characterize epidermal growth factor-related transforming growth factors in the urine of patients with hepatocellular carcinoma, gel filtration with Bio-Gel P-30 was performed in seven hepatocellular carcinoma patients and seven sex-matched and age-matched healthy controls. Distinct profiles of soft agar growth assay in the hepatocellular carcinoma patients and the normal controls were seen. Three peaks (A, B and C) in the urine were examined. Peak C in most hepatocellular carcinoma patients was higher than that in healthy controls. Similar profiles were detected with epidermal growth factor radioreceptor assay and cellular DNA synthesis assay. This result might indicate that transforming growth factors with low molecular weight were found in the urine of hepatocellular carcinoma patients. An exceptional HCC patient had an additional peak (A') that corresponded to the high molecular weight protein. We concluded that there were transforming growth factors with functional activity in the urine of patients with hepatocellular carcinoma.

Topics: Adult; Agar; Aged; Carcinoma, Hepatocellular; Cell Division; DNA, Neoplasm; Epidermal Growth Factor; ErbB Receptors; Humans; Liver Neoplasms; Middle Aged; Radioligand Assay; Transforming Growth Factors; Tumor Cells, Cultured

Earlier studies showed that the extracellular matrix and the conditioned medium from colon carcinoma support and catalyze the conversion of normal epithelial to colon carcinoma cells (14, 16). Since the cause of this apparent change in malignant potential is completely unknown, the following experiments examined molecular changes accompanying and mediating the transition. Colon epithelial cell cultures were initiated from normal colon biopsies. The cell cultures were carried on in standard medium on fibronectin-coated plates, and in conditioned medium on extracellular matrix from confirmed colon carcinoma. At time intervals, during 12 months period, aliquots were harvested, transferred into roller bottles to obtain enough cells to isolate cellular Poly(A)+-enriched RNA, cell membrane oligosaccharides and to determine cellular growth characteristics. During the 12 months in vitro culture, normal colon epithelial (NCE) cells grown on fibronectin in the standard growth medium maintained their initial characteristics. Whereas, NCE cells grown on the extracellular matrix and in colon carcinoma conditioned medium, progressively acquired the ability to form colonies in soft agar, to form tumors when implanted subcutaneously and to metastasize into the liver when administered intravenously into athymic mice. Poly(A)+-enriched RNA from NCE cells grown on fibronectin and in standard culture medium, did not, whereas the RNA from NCE cells grown on the extracellular matrix and in the colon carcinoma conditioned medium hybridized with 32P-cDNA from colon carcinoma. There were significant changes in the composition and profile of the oligosaccharides from membranes of NCE cells grown on the extracellular matrix. There was significant correlation (P less than 0.001) between the last characteristics.

Topics: Agar; Animals; Cell Membrane; Cells, Cultured; Colon; Colonic Neoplasms; Culture Media; Epithelial Cells; Epithelium; Female; Humans; Liver Neoplasms; Mice; Mice, Inbred BALB C; Neoplasm Metastasis; Oligosaccharides; Tumor Stem Cell Assay

A continuous human cell line was established from a patient with large cell anaplastic lung carcinoma. This cell line, designated QU-DB, has been in culture for over 36 months and grows as an adherent monolayer with a doubling time of 10-12 hours. Its morphology, ultrastructure, karyotype, ability to grow in soft agar and heterotransplantability, indicate it is a large-cell lung tumor cell line of human origin. Three cell lines were established from metastatic tumors in nude mice receiving subcutaneous injections of QU-DB cells. The morphology and growth characteristics exhibited by these cell lines were similar to the primary cell line. Karyotypic analysis of cell lines derived from the primary tumor and a metastasis to the diaphragm were similar, but cells from a liver metastasis culture showed additional karyotypic changes. This large cell lung tumor cell line may prove useful as a model system for studies of human tumor progression and metastasis.

Topics: Agar; Aged; Animals; Carcinoma; Cell Division; Cell Line; Clone Cells; Flow Cytometry; Humans; Karyotyping; Liver Neoplasms; Lung Neoplasms; Male; Mice; Mice, Nude; Microscopy, Electron; Neoplasm Metastasis; Smoking

Three parameters were evaluated as diagnostic of the malignant potential of cultured rat liver epithelial cells: cytology, growth in soft agar, and production of extracellular plasminogen activator. A total of 22 tumorigenic and nontumorigenic cultures from 15 cell lines were sent coded from their originators to two different laboratories for the evaluation of these three parameters. Cytologic diagnosis and growth in soft agar were reliable means of determining the malignant potential of the cultured cells. However, the production of extracellular plasminogen activator showed little correlation with tumorigenicity. Of cytologic properties evaluated, the two that correlated best with malignant potential were increased cytoplasmic basophilia and and increased nuclear:cytoplasmic ratio.

Topics: Agar; Animals; Cell Division; Cell Line; Cell Transformation, Neoplastic; Epithelium; Liver Neoplasms; Neoplasms, Experimental; Plasminogen Activators; Rats

Epithelial cultures established from adult rat liver and from rat hepatomas induced in vivo with aromatic amine carcinogens have been compared by light and electron microscopy and by growth properties in liquid medium and in agar. The morphology and growth patterns of all of these cultures indicate that they have characteristics of epithelial rather than fibroblast cells. The criteria generally used to score for transformation of fibroblasts were not satisfactory for distinguishing normal epithelial cells from hepatoma cells in culture. Growth in agar, however, provides a simple and objective method of scoring for transformed epithelial cells, because only the tumorigenic cells grow in agar. Since none of the normal cultures had hydrocortisone-inducible tryosine aminotransferase, we lack definitive evidence that they are derived from liver parenchymal cells. The outstanding feature in the ultrastructure of the hepatoma cells in culture was the presence of type A and C viral particles. Whereas five hepatoma cultures and a spontaneously transformed normal liver cell line were positive for these particles, five independently isolated cell cultures from normal adult rat liver were negative. Evidence is presented that the viral particles seen in hepatoma cultures are due to activation of latent viruses rather than to in vitro contamination. The possible significance of these particles in hepatocarcinogenesis is discussed.

Topics: Agar; Animals; Carcinogens; Carcinoma, Hepatocellular; Cell Division; Cell Transformation, Neoplastic; Cells, Cultured; Epithelial Cells; Epithelium; Liver; Liver Neoplasms; Microscopy, Electron; Neoplasms, Experimental; Oncogenic Viruses; Rats; Retroviridae

The sera of 445 persons, mainly patients with various liver diseases, were examined for alpha-fetoprotein (AFP) and hepatitis B antigens (HB Ag) and antibodies (HB Ab). The presence of AFP was detected in 9 sera by agar-gel precipitation and in 22 additional sera by the more sensitive aggregate haemagglutination technique. HB Ag proved to be present in 74 sera and HB Ab was present in 34 sera. Hepatitis B antigen was present more frequently (42%) in sera containing alpha-fetoprotein than in sera that did not contain it (14.7%). The food habits of the population investigated were also studied.

Topics: Agar; alpha-Fetoproteins; Antibodies, Viral; Chemical Precipitation; Feeding Behavior; Hemagglutination Tests; Hepatitis A; Hepatitis B Antigens; Humans; Immunoelectrophoresis; Liver Diseases; Liver Neoplasms; Mongolia

Topics: Agar; Animals; Carbon Isotopes; Carcinoma, Hepatocellular; Electrophoresis; Gels; Glycine; Isoenzymes; Liver; Liver Neoplasms; Mice; Mice, Inbred C3H; Neoplasms, Experimental; Proteins; Pyridoxal Phosphate; Rats; Serine; Spectrophotometry; Tetrahydrofolates; Transferases

Topics: Adenocarcinoma; Adenocarcinoma, Scirrhous; Adolescent; Adult; Agar; Aged; Carcinoma; Chromatography; Colonic Neoplasms; Gallbladder Neoplasms; Hemangiosarcoma; Humans; Liver Neoplasms; Lymphoma, Non-Hodgkin; Middle Aged; Neoplasm Proteins; Neoplasm Recurrence, Local; Pancreatic Neoplasms; Protein Denaturation; Retroperitoneal Neoplasms; Stomach Neoplasms

Epithelial-like cells originating from 10-day and 8-week old BD rats were treated in vitro with dimethylnitrosamine (DMN) and N-methyl-N'-nitro-nitrosoguanidine (MNNG). Morphologically, no differences were seen between treated and untreated cells up to the time when the cells were transplanted into syngeneic hosts. However, the treated cells grew in soft agar and once injected subcutaneously or intraperitoneally into newborn rats, developed tumours after a latent period of 9-12 weeks. The tumours obtained with DMN-treated cells were well differentiated adenocarcinomata, whereas carcinosarcomata were observed with the MNNG-treated cells.

Topics: Adenocarcinoma; Agar; Animals; Carcinoma; Cell Line; Epithelial Cells; Injections, Intraperitoneal; Injections, Subcutaneous; Liver Neoplasms; Neoplasm Transplantation; Nitrosamines; Nitrosoguanidines; Rats

Topics: Agar; Alloys; Aluminum; Animals; Arsenic; Carcinoma, Hepatocellular; Cell Adhesion; Cell Division; Cell Line; Densitometry; Ethylmaleimide; Glass; Iodoacetates; Kinetics; Liver Neoplasms; Mercuribenzoates; Mercury; Polyethylenes; Polymers; Polytetrafluoroethylene; Rats; Silicones

Topics: Agar; Electrophoresis; Gels; Hepatitis A; Humans; Isoenzymes; L-Lactate Dehydrogenase; Liver Neoplasms; Myocardial Infarction

Topics: Acyltransferases; Agar; Bile Duct Neoplasms; Biliary Tract Diseases; Cholelithiasis; Electrophoresis; Glutamates; Hepatitis; Humans; Isoenzymes; Jaundice; Liver Cirrhosis; Liver Diseases; Liver Neoplasms; Pancreatic Diseases; Pancreatic Neoplasms; Pancreatitis

Topics: Acute Disease; Agar; Alanine; Alkaline Phosphatase; Amides; Aminohydrolases; Anilides; Cholestasis; Chronic Disease; Electrophoresis; Hepatitis; Humans; Isoenzymes; Leucine; Liver Cirrhosis; Liver Diseases; Liver Neoplasms; Naphthalenes; Nitro Compounds; Pancreatic Diseases; Pancreatitis

Topics: Agar; Female; Fetal Proteins; Humans; Immunodiffusion; Liver Diseases; Liver Neoplasms; Spain

Topics: Agar; Animals; Anti-Bacterial Agents; Antineoplastic Agents; Ascites; Carcinoma, Hepatocellular; Clone Cells; Culture Techniques; Diffusion; Liver Neoplasms; Mercaptopurine; Methods; Mitomycins; Neoplasms, Experimental; Nitrogen Mustard Compounds; Rats; Thiotepa

Topics: Agar; Animals; Carcinoma; Carcinoma, Hepatocellular; Chemical Precipitation; Cricetinae; Electrophoresis; gamma-Globulins; In Vitro Techniques; Kidney; Liver Neoplasms; Neoplasm Proteins; Neoplasms, Experimental; Proteins; Quaternary Ammonium Compounds; Rats; Tissue Extracts

Topics: Agar; Alkaline Phosphatase; Bile; Bone Diseases; Bone Neoplasms; Child; Clinical Enzyme Tests; Electrophoresis; Hodgkin Disease; Humans; Jaundice; Jaundice, Obstructive; Liver Diseases; Liver Neoplasms; Lymphoma, Large B-Cell, Diffuse; Lymphoma, Non-Hodgkin; Osteitis Deformans; Sarcoma

Topics: Agar; Animals; Antigens; Carcinoma, Hepatocellular; Cytoplasm; Liver; Liver Neoplasms; Rats