tetracycline and Liver-Neoplasms

Patients with POAG have lower corneal endothelial cell density than healthy controls of the same age. This may be attributed to mechanical damage from elevated IOP and toxicity of glaucoma medications.. Mycophenolic acid was detected in all cats. The dose 10 mg/kg given q12h for 1 week was tolerated (n = 3). The efficacy of MMF as an immunosuppressant and long-term safety in cats of this dosage regimen is unknown.. T

Topics: Acetylcholine; Acinetobacter baumannii; Actinobacteria; Action Potentials; Adalimumab; Adaptation, Physiological; Adipates; Administration, Oral; Adolescent; Adrenal Glands; Adsorption; Adult; Aged; Aged, 80 and over; Aging; AIDS-Related Opportunistic Infections; Aldosterone; Amino Acids; Ammonia; Amoxicillin; AMP-Activated Protein Kinases; Animals; Antacids; Anti-Bacterial Agents; Antineoplastic Agents; Antirheumatic Agents; Apgar Score; Area Under Curve; ARNTL Transcription Factors; Arterial Pressure; Arthritis, Juvenile; Athletes; Attention; Biodegradation, Environmental; Biofilms; Biofuels; Biological Therapy; Biomass; Biomimetic Materials; Bioreactors; Birth Weight; Bismuth; Blood Flow Velocity; Bone and Bones; Brain Injuries, Traumatic; Calcium; Calcium Channels; Capsaicin; Carbon; Carcinoma, Hepatocellular; Cardiomegaly, Exercise-Induced; Cartilage; Cartilage, Articular; Case-Control Studies; Catalysis; Cats; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Death; Cell Differentiation; Cell Line, Tumor; Cell Membrane; Charcoal; Chemokine CCL2; Child; Child, Preschool; Chondrogenesis; Chronic Disease; Circadian Clocks; Circadian Rhythm Signaling Peptides and Proteins; Clarithromycin; Coccidioides; Coccidioidomycosis; Cognitive Behavioral Therapy; Coinfection; Color; Coloring Agents; Computer Simulation; Computers, Molecular; Consensus; Corticosterone; Cyclic AMP Response Element-Binding Protein; Cytochrome P-450 Enzyme System; Death, Sudden, Cardiac; Density Functional Theory; Diabetes Mellitus, Type 2; Diabetic Retinopathy; Dialysis Solutions; Disease Models, Animal; Dogs; Dopamine Agonists; Dose-Response Relationship, Drug; Doxorubicin; Drug Administration Schedule; Drug Resistance, Bacterial; Drug Therapy, Combination; Electrocardiography; Electrocardiography, Ambulatory; Electrolytes; Endocardium; Endocrine Disruptors; Endocytosis; Endoscopy, Gastrointestinal; Escherichia coli Proteins; Esters; Evolution, Molecular; Executive Function; Feasibility Studies; Female; Ferric Compounds; Fluorescence; Fluorescent Dyes; Fluorine Radioisotopes; Frailty; Free Radical Scavengers; Gabapentin; Geriatric Assessment; Glucaric Acid; Glucocorticoids; Glucose; Glucose Metabolism Disorders; Halogenated Diphenyl Ethers; Heart Rate; Heart Ventricles; HEK293 Cells; Helicobacter Infections; Helicobacter pylori; Hep G2 Cells; Hepatocytes; Humans; Hungary; Hydrogen Sulfide; Hydrogen-Ion Concentration; Immunologic Factors; Immunomodulation; Immunosuppressive Agents; Independent Living; Indocyanine Green; Infant; Infant Formula; Infant Mortality; Infant, Newborn; Infant, Newborn, Diseases; Inflorescence; Insulin Resistance; Insulins; International Agencies; Iron; Isotonic Solutions; Kidney Failure, Chronic; Kinetics; Lactones; Leukocytes, Mononuclear; Liver Neoplasms; Macular Edema; Magnetic Resonance Imaging; Magnetic Resonance Spectroscopy; Magnetosomes; Male; Medical Audit; Mesenchymal Stem Cells; Metabolic Syndrome; Metformin; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Nude; Middle Aged; Molecular Conformation; Molecular Targeted Therapy; Motor Activity; Multiple Sclerosis; Mycophenolic Acid; Netherlands; Neuropsychological Tests; Nuclear Energy; Organs at Risk; Osteoarthritis; Osteoarthritis, Hip; Oxidation-Reduction; Palladium; Pericardium; Perinatal Death; Peritoneal Dialysis; Phantoms, Imaging; Pharmaceutical Preparations; Phospholipids; Phosphorylation; Physical Conditioning, Human; Physical Endurance; Pilot Projects; Polyketides; Polymers; Positron-Emission Tomography; Postoperative Period; Potassium; Powders; Pramipexole; Predictive Value of Tests; Pregabalin; Pregnancy; Pregnancy Outcome; Protein Structure, Secondary; Proton Pump Inhibitors; Puberty; Pulmonary Circulation; Quality Assurance, Health Care; Quantum Dots; Radiometry; Radiotherapy Dosage; Radiotherapy Planning, Computer-Assisted; Radiotherapy, Intensity-Modulated; Rats, Sprague-Dawley; Receptors, CCR2; Receptors, Transferrin; Regeneration; Registries; Renal Insufficiency, Chronic; Reproducibility of Results; Research Design; Restless Legs Syndrome; Retina; Retinoid X Receptor alpha; Retrospective Studies; Rhenium; Risk Factors; RNA, Messenger; Severity of Illness Index; Sex Factors; Sodium; Sodium Fluoride; Solvents; Spectrometry, Fluorescence; Spectroscopy, Fourier Transform Infrared; Stereoisomerism; Stroke; Structure-Activity Relationship; Tachycardia, Ventricular; Tetracycline; Tetrahydrofolate Dehydrogenase; Tetrahydronaphthalenes; Thermodynamics; Thiophenes; Time Factors; Tinidazole; Tomography, Optical Coherence; Tomography, X-Ray Computed; Topiramate; Toxoplasma; Toxoplasmosis, Cerebral; Transferrin; Treatment Outcome; Up-Regulation; Upper Extremity; Uremia; Uveitis; Vascular Remodeling; Ventricular Fibrillation; Ventricular Function, Left; Ventricular Function, Right; Ventricular Remodeling; Verapamil; Veterans; Visual Acuity; Vitrectomy; Water Pollutants, Chemical; Zea mays; Zirconium

Anatomic pathology has produced considerable knowledge about environmental teratogens and carcinogens. A special disease registry established by a pathologist provided details of the association between oral contraceptives and hepatic neoplams. Pathologists were also involved in establishing in the link between diethylstilbestrol use and clear-cell adenocarcinomas of the vagina. An area of particular interest has been gender and ethnic differences in the incidence of certain diseases. Pathologists further make use of animal studies to investigate the pathogenesis of human tumors. Finally, stored serum or tissue is often used by pathologists to help diagnose diseases retrospectively. Human skin fibroblasts grown in culture and stored have been especially valuable for laboratory research. This chapter briefly highlights some of the milestones in the detection of enviromental effects through anatomic pathology.

Topics: Adenocarcinoma; Animals; Ataxia Telangiectasia; Contraceptives, Oral; Diethylstilbestrol; Disease Models, Animal; Environment; Ethnicity; Female; Geography; Herpesvirus 4, Human; Humans; Liver Neoplasms; Lymphoproliferative Disorders; Osteosarcoma; Pathology, Clinical; Radiation, Ionizing; Sex Factors; Tetracycline; Thorium Dioxide; Tooth Discoloration; Vaginal Neoplasms; X Chromosome

Topics: Biliary Tract; Carcinoma, Hepatocellular; Humans; Liver; Liver Neoplasms; Organotechnetium Compounds; Pyrrolidines; Radionuclide Imaging; Spleen; Technetium Tc 99m Aggregated Albumin; Technetium Tc 99m Pentetate; Tetracycline

High levels of S100A6 have been associated with poor outcome in some types of human cancers, but the role of S100A6 in the molecular pathogenesis of these cancers is largely unknown. This study was performed to explore the expression and functional roles of S100A6 in hepatocellular carcinoma (HCC). The expression level of S100A6 in HCC tumor and corresponding peritumoral tissues were determined by immunohistochemistry analysis. The potential functions of S100A6 in tumorigenesis and metastasis were analyzed by cell proliferation, migration, and invasion assays in human liver cancer cells. Moreover, through expression and purification of S100A6 recombinant protein tagged with cell-penetrating peptide, we analyzed its complex extracellular/intracellular effects in a S100A6-silenced cellular model. As a result, the expression of S100A6 was up-regulated in human HCC compared with adjacent peritumoral tissues. S100A6 silencing inhibited the growth and motility of HCC cells, while intracellular re-expression of S100A6 could rescue the proliferation and migration defects. Intracellular over-expression of S100A6 resulted in down-regulation of E-cadherin expression and promoted nuclear accumulation of β-catenin. Moreover, we found that the enhanced cell proliferation and motility after S100A6 stimulation were dependent on the activation of PI3K/AKT pathway. These results suggest that S100A6 may be involved in promotion and progression of human liver cancer.. S100A6 is overexpressed in human hepatocellular carcinoma clinical specimens. S100A6 promotes proliferation and migration of human hepatoma cells. Overexpression of S100A6 results in alteration of E-cadherin and β-catenin. The multi-effects of S100A6 may be mediated in part by PI3K/AKT pathway activation.

Topics: Animals; beta Catenin; Cadherins; Carcinogenesis; Carcinoma, Hepatocellular; Cell Cycle Proteins; Cell Movement; Cell Nucleus; Cell Proliferation; Cell Shape; Cholangiocarcinoma; Down-Regulation; G1 Phase Cell Cycle Checkpoints; Gene Silencing; Hep G2 Cells; Humans; Immunohistochemistry; Liver Neoplasms; Mice; Phosphatidylinositol 3-Kinases; Protein Transport; Proto-Oncogene Proteins c-akt; Pseudopodia; S100 Calcium Binding Protein A6; S100 Proteins; Signal Transduction; Tetracycline

Myc is a pleiotropic transcription factor that is involved in many cellular activities relevant to carcinogenesis, including hepatocarcinogenesis. The zebrafish has been increasingly used to model human diseases and it is particularly valuable in helping to identify common and conserved molecular mechanisms in vertebrates. Here we generated a liver tumor model in transgenic zebrafish by liver-specific expression of mouse Myc using a Tet-On system. Dosage-dependent induction of Myc expression specifically in the liver was observed in our Myc transgenic zebrafish, TO(Myc), and the elevated Myc expression caused liver hyperplasia, which progressed to hepatocellular adenoma and carcinoma with prolonged induction. Next generation sequencing-based transcriptomic analyses indicated that ribosome proteins were overwhelmingly upregulated in the Myc-induced liver tumors. Cross-species analyses showed that the zebrafish Myc model correlated well with Myc transgenic mouse models for liver cancers. The Myc-induced zebrafish liver tumors also possessed molecular signatures highly similar to human those of hepatocellular carcinoma. Finally, we found that a small Myc target gene set of 16 genes could be used to identify liver tumors due to Myc upregulation. Thus, our zebrafish model demonstrated the conserved role of Myc in promoting hepatocarcinogenesis in all vertebrate species.

Topics: Animals; Animals, Genetically Modified; Carcinoma, Hepatocellular; Disease Models, Animal; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Humans; Liver; Liver Neoplasms; Liver Neoplasms, Experimental; Mammals; Mice; Mice, Transgenic; Neoplasm Staging; Proto-Oncogene Proteins c-myc; Reproducibility of Results; Sequence Analysis, RNA; Tetracycline; Zebrafish

We report a 62-year-old man who presented with pain on the right side of his hip. CT revealed destructive masses in the right femur and left ilium. Histological examination indicated metastases from hepatocellular carcinoma, and further investigations revealed the primary tumor in the liver. Hepatobiliary scintigraphy using 99mTc N-pyrydoxyl-5-methyltryptophan and fused SPECT/CT clearly showed abnormal accumulation in these bone metastases from hepatocellular carcinoma.

Topics: Biliary Tract; Bone Neoplasms; Carcinoma, Hepatocellular; Humans; Liver; Liver Neoplasms; Male; Middle Aged; Multimodal Imaging; Organotechnetium Compounds; Positron-Emission Tomography; Pyrrolidines; Tetracycline; Tomography, X-Ray Computed

Recombinant adeno-associated virus (rAAV) are effective gene delivery vehicles that can mediate long-lasting transgene expression. However, tight regulation and tissue-specific transgene expression is required for certain therapeutic applications. For regulatable expression from the liver we designed a hepatospecific bidirectional and autoregulatory tetracycline (Tet)-On system (Tet(bidir)Alb) flanked by AAV inverted terminal repeats (ITRs). We characterized the inducible hepatospecific system in comparison with an inducible ubiquitous expression system (Tet(bidir)CMV) using luciferase (luc). Although the ubiquitous system led to luc expression throughout the mouse, luc expression derived from the hepatospecific system was restricted to the liver. Interestingly, the induction rate of the Tet(bidir)Alb was significantly higher than that of Tet(bidir)CMV, whereas leakage of Tet(bidir)Alb was significantly lower. To evaluate the therapeutic potential of this vector, an AAV-Tet(bidir)-Alb-expressing interleukin-12 (IL-12) was tested in a murine model for hepatic colorectal metastasis. The vector induced dose-dependent levels of IL-12 and interferon-γ (IFN-γ), showing no significant toxicity. AAV-Tet(bidir)-Alb-IL-12 was highly efficient in preventing establishment of metastasis in the liver and induced an efficient T-cell memory response to tumor cells. Thus, we have demonstrated persistent, and inducible in vivo expression of a gene from a liver-specific Tet-On inducible construct delivered via an AAV vector and proved to be an efficient tool for treating liver cancer.

Topics: Alanine Transaminase; Animals; Aspartate Aminotransferases; Cell Line; Dependovirus; Doxycycline; Female; Flow Cytometry; Gene Expression Regulation, Neoplastic; Genetic Therapy; Genetic Vectors; HEK293 Cells; HeLa Cells; Hep G2 Cells; Humans; Interferon-gamma; Interleukin-12; Liver Neoplasms; Male; Mice; Mice, Inbred BALB C; Tetracycline

Drug-induced liver injury occurs in general after several weeks and is often unpredictable. It is characterized by a large spectrum of lesions that includes steatosis and phospholipidosis. Many drugs such as amiodarone and tetracycline have been reported to cause phospholipidosis and/or steatosis. In this study, acute and chronic hepatic effects of these two drugs were investigated using well-differentiated human hepatoma HepaRG cells. Accumulation of typical lipid droplets, labeled with Oil Red O, was observed in hepatocyte-like HepaRG cells after repeat exposure to either drug. Amiodarone caused the formation of additional intracytoplasmic vesicles that did not stain in all HepaRG cells. At the electron microscopic level, these vesicles appeared as typical lamellar bodies and were associated with an increase of phosphatidylethanolamine and phosphatidylcholine. A dose-dependent induction of triglycerides (TG) was observed after repeat exposure to either amiodarone or tetracycline. Several genes known to be related to lipogenesis were induced after treatment by these two drugs. By contrast, opposite deregulation of some of these genes (FASN, SCD1, and THSRP) was observed in fat HepaRG cells induced by oleic acid overload, supporting the conclusion that different mechanisms were involved in the induction of steatosis by drugs and oleic acid. Moreover, several genes related to lipid droplet formation (ADFP, PLIN4) were up-regulated after exposure to both drugs and oleic acid.. Our results show that amiodarone causes phospholipidosis after short-term treatment and, like tetracycline, induces vesicular steatosis after repeat exposure in HepaRG cells. These data represent the first demonstration that drugs can induce vesicular steatosis in vitro and show a direct relationship between TG accumulation and enhanced expression of lipogenic genes.

Topics: Amiodarone; Carcinoma, Hepatocellular; Carrier Proteins; Cell Line, Tumor; Down-Regulation; Fatty Acid Synthase, Type I; Fatty Liver; Humans; Lipid Metabolism; Lipogenesis; Liver Neoplasms; Membrane Proteins; Oleic Acid; Perilipin-2; Perilipin-4; Phosphoproteins; Stearoyl-CoA Desaturase; Tetracycline; Triglycerides; Up-Regulation

Hepatitis C virus (HCV) is a significant health problem facing the world. More than 170 million people are infected with HCV worldwide. HCV encodes a large polyprotein precursor that is processed into at least 10 distinct products including structural (core, E1 and E2) and non-structural (NS2, NS3, NS4A, NS4B, NS5A and NS5B). Besides its importance in virus replication, NS4A functions as a cofactor for NS3 and contributes to viral pathogenesis by influencing cellular functions. Here, we investigated the effect of NS4A protein on the growth rate induced by core protein in liver cells. Using our established tetracycline inducible system, we demonstrated the ability of NS4A protein to inhibit core protein-induced cell growth in Hepatoma cell line, HepG2. Induction of both core and NS4A proteins in HepG2-core/NS4A transfectants inhibited core-induced growth advantage in HepG2-core transfectants and blocked NS4A protein-induced cell growth inhibition in HepG2-NS4A transfectants. Using both immune fluorescence staining and Western blot analysis, we confirmed the localization of NS4A protein to the mitochondria in HepG2-NS4A transfectants expressing NS4A protein. Data obtained from flow cytometry analysis, using JC-1 demonstrated the loss of mitochondrial membrane potential (DeltaPsim) by the expression of NS4A protein in HepG2-NS4A transfectants, but not by the expression of core protein in HepG2-core transfectants. Whereas, the induction of the expression of both core and NS4A proteins in HepG2-core/NS4A transfectants blocked NS4A-induced loss of DeltaPsim in HepG2 cells. Taken together, our data suggest an important role for mitochondria in the modulation HCV NS4A-induced inhibition of HCV core-mediated cell growth.

Topics: Carcinoma, Hepatocellular; Carrier Proteins; Cell Line, Tumor; Cell Proliferation; Gene Expression Regulation, Viral; Hepacivirus; Humans; Intracellular Signaling Peptides and Proteins; Liver Neoplasms; Membrane Potential, Mitochondrial; Mitochondria, Liver; Protein Synthesis Inhibitors; Tetracycline; Viral Core Proteins; Viral Nonstructural Proteins; Viral Proteins

Induction of p53 gene expression in cancer cells can lead to both cell cycle arrest and apoptosis. To clarify whether the level of p53 expression determines the apoptotic response of hepatocellular carcinoma (HCC) cells, we assessed the effect of various levels of expression of p53 gene on a p53-deficient HCC cell line, Hep3B, utilizing a doxycycline (Dox)-regulated inducible p53 expression system. Our results showed that apoptosis was induced in HCC cells with high levels of p53 expression. However, lower level of p53 expression induced only cell cycle arrest but not apoptosis. Bax expression was up-regulated following high levels of p53 expression, while bcl-2 expression was not altered by the level of p53 expression. Moreover, p21 expression was observed in both high and low expression of p53. These results suggest the level of p53 expression could determine if the HCC cells would go into cell cycle arrest or apoptosis. Bax may participate, at least in part, in inducing p53-dependent apoptosis and the induction of p21 alone was able to cause cell cycle arrest but not apoptosis.

Topics: Anti-Bacterial Agents; Apoptosis; bcl-2-Associated X Protein; Carcinoma, Hepatocellular; Cell Cycle; Cell Line, Tumor; Clone Cells; Cyclin-Dependent Kinase Inhibitor p21; Doxycycline; Gene Expression Regulation, Neoplastic; Humans; Liver Neoplasms; Tetracycline; Tumor Suppressor Protein p53; Up-Regulation

The hyaluronan receptor CD44 plays an important role in facilitating invasion and metastasis of a variety of tumors, including breast carcinomas. CD44 functions as a bioactive signaling transmitter. Although a number of studies have implicated CD44 in breast tumor invasion, the evidence is still circumstantial. We have developed a tetracycline-regulated CD44s (standard form) system in the weakly metastatic breast cancer cell MCF7, which exhibits low endogenous expression of CD44 and generated a new cell line, MCF7F-B5. Induction of CD44s alone affected the growth characteristics of MCF7F-B5 cells by increasing their abilities to proliferate, migrate, and invade in vitro. In addition, we have identified and validated cortactin as a novel transcriptional target of hyaluronan/CD44s signaling in underpinning breast tumor invasion. To test these observations in vivo, we developed a doxycycline (DOX)-regulated CD44s breast cancer xenograft model. Induction of CD44s did not affect the growth rate or local invasion of the primary tumor. However, although no mice from the +DOX group developed metastasis, 8 of 11 mice from the -DOX group developed secondary tumors to the liver only. Interestingly, metastatic breast tumors expressed high levels of CD44. This study provides in vivo evidence for the role of the standard form of CD44 in promoting breast tumor invasion and metastasis to the liver.

Topics: Animals; Breast Neoplasms; Cell Line, Tumor; Disease Models, Animal; Doxycycline; Female; Hyaluronan Receptors; Liver Neoplasms; Mice; Mice, SCID; Neoplasm Invasiveness; Neoplasm Transplantation; Tetracycline; Transplantation, Heterologous

Precise control of the level of protein expression in cells can yield quantitative and temporal information on the role of a given gene in normal cellular physiology and on exposure to chemicals and drugs. This is particularly relevant to liver cells, in which the expression of many proteins, such as phase I and phase II drug-metabolizing enzymes, vary widely between species, among individual humans, and on exposure to xenobiotics. The most widely used gene regulatory system has been the tet-on/off approach. Although a second-generation tet-on transactivator was recently described, it has not been widely investigated for its potential as a tool for regulating genes in cells and particularly in cells previously recalcitrant to the first-generation tet-on approach, such as hepatocyte-derived cells. Here we demonstrate the development of two human (HepG2 and HuH7) and one mouse (Hepa1c1c7) hepatoma-derived cell lines incorporating a second-generation doxycycline-inducible gene expression system and the application of the human lines to control the expression of different transgenes. The two human cell lines were tested for transient or stable inducibility of five transgenes relevant to liver biology, namely phase I (cytochrome P-450 2E1; CYP2E1) and phase II (glutathione S-transferase P1; GSTP1) drug metabolism, and three transcription factors that respond to chemical stress [nuclear factor erythroid 2 p45-related factors (NRF)1 and 2 and NFKB1 subunit of NF-kappaB]. High levels of functional expression were obtained in a time- and dose-dependent manner. Importantly, doxycycline did not cause obvious changes in the cellular proteome. In conclusion, we have generated hepatocyte-derived cell lines in which expression of genes is fully controllable.

Topics: Amino Acid Sequence; Animals; Anti-Bacterial Agents; Carcinoma, Hepatocellular; Cell Line, Tumor; Cytochrome P-450 CYP2E1; Doxycycline; Gene Expression Regulation, Neoplastic; Glutathione Transferase; Hepatocytes; Humans; Liver; Liver Neoplasms; Mice; Molecular Sequence Data; NF-E2-Related Factor 1; NF-E2-Related Factor 2; NF-kappa B p50 Subunit; Protein Synthesis Inhibitors; Tetracycline; Transgenes; Xenobiotics

The hepatitis B virus (HBV) X protein (pX) is implicated in hepatocarcinogenesis by an unknown mechanism. pX variants encoded by HBV genomes found integrated in genomic DNA from liver tumors of patients with hepatocellular carcinoma (HCC) generally lack amino acids 134 to 154. Since deregulation of mitogenic pathways is linked to oncogenic transformation, herein we define the pX region required for mitogenic pathway activation. A series of pX deletions was used to construct tetracycline-regulated pX-expressing cell lines. The activation of the mitogenic pathways by these pX deletions expressed in the constructed cell lines was measured by transient transreporter assays, effects on endogenous cyclin A expression, and apoptosis. Conditional expression of pX51-140 in AML12 clone 4 cell line activates the mitogenic pathways, induces endogenous cyclin A expression, and sensitizes cells to apoptosis, similar to wild-type (WT) pX. By contrast, pX1-115 is inactive, supporting the idea that amino acids 116 to 140 are required for mitogenic pathway activation. Moreover, this pX deletion analysis demonstrates that WT pX function is modulated by two regions spanning amino acids 1 to 78 and 141 to 154. The N-terminal X1-78, expressed via a retroviral vector in WT pX-expressing 4pX-1 cells, coimmunoprecipitates with WT pX, indicating this pX region participates in protein-protein interactions leading to pX oligomerization. Interestingly, pX1-78 interferes with WT pX in mediating mitogenic pathway activation, endogenous gene expression, and apoptosis. The C-terminal pX region spanning amino acids 141 to 154 decreases pX stability, determined by pulse-chase studies of WT pX and pX1-140, suggesting that increased stability of naturally occurring pX variants lacking amino acids 134 to 154 may play a role in HCC development.

Topics: Animals; Apoptosis; Carcinoma, Hepatocellular; Cell Line; Cyclin A; Gene Expression; Genes, Viral; Hepatitis B virus; Humans; Liver Neoplasms; Mice; Mitogens; Recombinant Proteins; Sequence Deletion; Signal Transduction; Tetracycline; Trans-Activators; Transfection; Viral Regulatory and Accessory Proteins

We report a case of a 61-year-old man who presented with fatigue, abdominal pain and hepatomegaly. Computed tomography (CT) of the abdomen showed hepatomegaly and multiple hepatic lesions highly suggestive of metastatic diseases. Due to the endoscopic finding of colon ulcer, colon cancer with liver metastases was suspected. Biochemically a slight increase of transaminases, alkaline phosphatase and gammaglutamyl transpeptidase were present; alpha-fetoprotein, carcinoembryogenic antigen and carbohydrate 19-9 antigen serum levels were normal. Laboratory and instrumental investigations, including colon and liver biopsies revealed no signs of malignancy. In the light of spontaneous improvement of symptoms and CT findings, his personal history was reevaluated revealing direct contact with pigs and their tissues. Diagnosis of leptospirosis was considered and confirmed by detection of an elevated titer of antibodies to leptospira. After two mo, biochemical data, CT and colonoscopy were totally normal.

Topics: Colonic Neoplasms; Colonoscopy; Contrast Media; Diagnosis, Differential; Gram-Negative Bacteria; Humans; Leptospirosis; Liver Function Tests; Liver Neoplasms; Male; Middle Aged; Tetracycline; Tomography, X-Ray Computed

We describe here a case in which 99mTc-Sn-N-pyridoxy-5-methyltryptophan (99mTc-PMT) scintigraphy was useful in diagnosing renal metastasis of hepatocellular carcinoma (HCC). A 62-year-old man who had undergone hepatectomy for HCC presented 6 years after initial diagnosis with left flank pain and was found on CT and MRI to have a tumor in the left kidney. Hepatobiliary scintigraphy using 99Tc-PMT was performed, and 99mTc-PMT accumulation was found in the tumor. Nephrectomy was performed and metastasis of HCC was confirmed.

Topics: Carcinoma, Hepatocellular; Humans; Kidney Neoplasms; Liver Neoplasms; Male; Middle Aged; Organotechnetium Compounds; Pyrrolidines; Radionuclide Imaging; Radiopharmaceuticals; Tetracycline

We cloned the complete complementary DNA of an isolate of the hepatitis C virus, HCV-S1, into a tetracycline-inducible expression vector and stably transfected it into two human hepatoma cell lines, Huh7 and HepG2. Twenty-six Huh7 and two HepG2-positive clones were obtained after preliminary screening. Two Huh7 (SH-7 and -9) and one HepG2 (G-19) clones were chosen for further characterisation. Expression of HCV proteins in these cells accumulated from 6 h to 4 days posttreatment. Full-length viral plus-strand RNA was detected by Northern analyses. Using RT-PCR and ribonuclease protection assay, we also detected the synthesis of minus-strand HCV RNA. Plus- and minus-strand viral RNA was still detected after treatment with actinomycin D. Indirect immunofluorescence staining with anti-E2, NS4B, and NS5A revealed that these proteins were mostly localised to the endoplasmic reticulum (ER). Culture media from tet-induced SH-9 cells was separated on sucrose density gradients and analysed for the presence of HCV RNA. Viral RNA levels peaked at two separate ranges, one with a buoyant density of 1.08 g/ml and another from 1.17 to 1.39 g/ml. Electron microscopy demonstrated the presence of subviral-like particles (approximately 20-25 nm in diameter) in the cytoplasm of SH-9 and G-19 cells, which were positively labelled by anti-HCV core antibodies. Anti-E2 antibodies strongly labelled cytoplasmic vesicular structures and some viral-like particles. Complete viral particles of about 50 nm which reacted with anti-E2 antibodies were observed in the culture media of tet-induced SH-9 cells following negative staining. Supernatant from tet-treated SH-9 cells was found to infect nai;ve Huh7 and stable Huh7-human CD81 cells.

Topics: Carcinoma, Hepatocellular; Centrifugation, Density Gradient; Culture Media; Endoplasmic Reticulum; Genetic Vectors; Hepacivirus; Hepatitis C Antigens; Humans; Liver Neoplasms; RNA, Viral; Tetracycline; Transfection; Tumor Cells, Cultured; Viral Core Proteins; Viral Envelope Proteins; Viral Nonstructural Proteins; Viral Proteins; Virion; Virus Replication

The growth of any solid tumor depends on angiogenesis. Among the known angiogenic factors, basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), are potent and representative factors involved in tumor development. It has been reported that bFGF and VEGF showed a synergistic effect in both in vitro and in vivo angiogenesis. However, the interaction of these factors on tumor development and angiogenesis, including hepatocellular carcinoma (HCC), has not yet been elucidated. In this study, we examined the combined effect of bFGF and VEGF overexpression by means of a combination of a retroviral tetracycline (tet)-regulated (Retro-Tet) gene expression system, which can manipulate the gene expression in vivo by providing tet in the drinking water, and a conventional plasmid gene expression system. In an allograft study, bFGF and VEGF overexpression synergistically increased tumor growth and angiogenesis in the murine HCC cells. This synergistic effect also was found in established tumors. VEGF messenger RNA (mRNA) expression in the tumor was increased 3.1-fold by bFGF-overexpression, and the bFGF-induced tumor development was significantly attenuated by treatment with KDR/Flk-1 neutralizing monoclonal antibody. In conclusion, these results suggest that bFGF synergistically augments VEGF-mediated HCC development and angiogenesis at least partly by induction of VEGF through KDR/Flk-1.

Topics: Animals; Carcinoma, Hepatocellular; Drug Synergism; Endothelial Growth Factors; Fibroblast Growth Factor 2; Gene Transfer Techniques; Genetic Vectors; Injections, Subcutaneous; Liver Neoplasms; Lymphokines; Mice; Mice, Inbred BALB C; Neoplasm Transplantation; Neovascularization, Pathologic; Protein Synthesis Inhibitors; Receptor Protein-Tyrosine Kinases; Receptors, Growth Factor; Receptors, Vascular Endothelial Growth Factor; Retroviridae; Tetracycline; Tumor Cells, Cultured; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

Adaptive epigenetic changes and toxicity often accompany constitutive expression of a transgene or knockout of an endogenous gene in mice. These considerations potentially limit the usefulness of transgenic technology in studying the in vivo functions of a gene. Using conditional gene expression technology, it is possible to override such restrictions to achieve temporal and tissue-specific manipulation of gene expression in vivo. Based on the tetracycline regulatory system, we established a binary transgenic model in which the conditional expression of two transgenes, SV40 T antigen (TAg) and lacZ, can be tightly regulated in the liver by administration of tetracycline. The mouse albumin or mouse major urinary protein promoter was used to achieve liver-specific expression of the tetracycline-responsive transcriptional activator (tTA) in one set of transgenic mice. These mice were crossed with transgenic mice carrying either TAg or lacZ under the control of the tTA-regulated promoter. Analyses of mice transgenic for both tTA and TAg (or lacZ) revealed that the liver-specific expression of the transgenes could be suppressed to undetectable levels and regulated in a reversible fashion by tetracycline administration and withdrawal. Mice with tTA and TAg transgenes developed hepatocellular adenomas and hyperplasia that could be prevented by continuous tetracycline administration. Our report demonstrates the value of this binary transgenic model in studying the physiological functions of any potential genes of interest in a liver-specific manner.

Topics: Adenoma; Albumins; Animals; Anti-Bacterial Agents; Antigens, Polyomavirus Transforming; Crosses, Genetic; Gene Expression Regulation; Gene Transfer Techniques; Genotype; Hyperplasia; Lac Operon; Liver; Liver Neoplasms; Mice; Mice, Transgenic; Promoter Regions, Genetic; Proteins; Spleen; Tetracycline; Transcriptional Activation; Transgenes

Topics: Bone Neoplasms; Carcinoma, Hepatocellular; Clavicle; Humans; Ilium; Liver Neoplasms; Male; Middle Aged; Organotechnetium Compounds; Pyrrolidines; Radionuclide Imaging; Radiopharmaceuticals; Ribs; Scapula; Skull; Technetium Tc 99m Medronate; Tetracycline

Tc-99m PMT and Tc-99m GSA can be taken up by hepatocellular carcinoma (HCC), but there has been no report concerning HCC showing accumulation of both of Tc-99m PMT and Tc-99m GSA. In this paper we describe a case of two simultaneously developed HCCs, one of which took up both tracers but the other took up neither of them.

Topics: Aged; Carcinoma, Hepatocellular; Female; Humans; Liver Neoplasms; Magnetic Resonance Imaging; Organotechnetium Compounds; Pyrrolidines; Radionuclide Imaging; Radiopharmaceuticals; Reproducibility of Results; Technetium Tc 99m Aggregated Albumin; Technetium Tc 99m Pentetate; Tetracycline; Tissue Distribution; Tomography, X-Ray Computed

Angiogenesis is essential for the development of a solid tumor, including hepatocellular carcinoma (HCC). HCC is a well-known hypervascular tumor. Vascular endothelial growth factor (VEGF) is one of the most potent angiogenic factors. Its role has not been clarified in vivo in HCC development. We used a self-contained, tetracycline-regulated retroviral vector system to elucidate the effect of VEGF on murine HCC development in a xenograft experimental model. By delivering the VEGF gene within the retroviral vector and under the control of a tetracycline-regulated promoter, we were able to manipulate VEGF expression in vivo tumor by providing tetracycline in the drinking water. Overexpression of VEGF showed a marked increase in tumor development accompanied by augmentation of neovascularization. The degree of tumor enlargement corresponded to the level of VEGF gene expression. Suppression of VEGF led to a decrease in tumor growth at the established tumor size, whether relatively small or large. The level of VEGF expression did not alter the proliferation of HCC cells in vitro. In a double-chamber chemoinvasion assay, the in vitro invasion activity of VEGF-transduced cells was not changed. In the presence of endothelial cells (EC), however, VEGF-transduced cells showed a marked increase in their in vitro invasion activity. These results suggested that VEGF plays a critical role in the development of HCC in cooperation with EC

Topics: Animals; Carcinoma, Hepatocellular; Cell Division; Cell Movement; Endothelial Growth Factors; Endothelium, Vascular; Gene Expression Regulation; Genetic Vectors; Humans; Liver Neoplasms; Lymphokines; Mice; Neoplasm Invasiveness; Platelet Endothelial Cell Adhesion Molecule-1; Protein Synthesis Inhibitors; Retroviridae; Tetracycline; Tumor Cells, Cultured; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

We report the development and isolation of a cell line, termed HepAD38, that replicates human hepatitis B virus (HBV) under conditions that can be regulated with tetracycline. In the presence of the antibiotic, this cell line is free of virus due to the repression of pregenomic (pg) RNA synthesis. Upon removal of tetracycline from the culture medium, the cells express viral pg RNA, accumulate subviral particles in the cytoplasm that contain DNA intermediates characteristic of viral replication, and secrete virus-like particles into the supernatant. Since the HepAD38 cell line can produce high levels of HBV DNA, it should be useful for analyses of the viral replication cycle that depend upon viral DNA synthesis in a synchronized fashion. In addition, this cell line has been formatted into a high-throughput, cell-based assay that permits the large-scale screening of diverse compound libraries for new classes of inhibitors of HBV replication.

Topics: Cell Line; DNA, Viral; Hepatitis B virus; Hepatoblastoma; Humans; Liver Neoplasms; Protein Synthesis Inhibitors; RNA, Viral; Tetracycline; Transfection; Tumor Cells, Cultured; Virus Replication

The diagnostic value of whole body scanning using 99mTc-N-pyridoxylmethyltryptophan (PMT) was evaluated in 16 patients with bone metastases from hepatocellular carcinoma, in comparison with 99mTc-MDP. Of the 72 known lesions of bone metastases, 63 (87.5%) were detected by 99mTc-PMT scintigraphy, which demonstrated increased uptake of radionuclide. However, 99mTc-MDP bone scintigraphy detected only 45 lesions (62.5%), which were shown as increased, decreased, or mixed patterns of uptake. Thus 99mTc-PMT scintigraphy was more sensitive than 99mTc-MDP bone scintigraphy. In addition, the latter showed poor specificity because of its high false positive rate due to degenerative change. All lesions undetected by 99mTc-PMT scintigraphy were located in areas that overlapped the liver or bowel activity. In conclusion, it is recommended that whole body 99mTc-PMT scintigraphy be combined with 99mTc-MDP bone scintigraphy for the detection of bone metastases from hepatocellular carcinoma.

Topics: Aged; Aged, 80 and over; Bone Neoplasms; Carcinoma, Hepatocellular; Female; Humans; Japan; Liver Neoplasms; Male; Middle Aged; Organotechnetium Compounds; Pyrrolidines; Radionuclide Imaging; Retrospective Studies; Tetracycline; Tetracyclines

The authors report discordant hepatic uptake of Tc-99m NGA and Tc-99m PMT in a patient with hepatoma. Tc-99m PMT uptake was delayed and Tc-99m NGA concentrated in another area, thereby demonstrating that the uptake mechanisms for Tc-99m NGA and Tc-99m PMT are different. Tc-99m NGA imaging may be useful in characterizing the focal hepatic lesion of Tc-99m IDA or Tc-99m PMT concentration.

Topics: Albumins; Carcinoma, Hepatocellular; Humans; Liver Neoplasms; Male; Middle Aged; Organotechnetium Compounds; Pyrrolidines; Radionuclide Imaging; Tetracycline; Tetracyclines

Topics: Adrenal Gland Neoplasms; Aged; Aged, 80 and over; Carcinoma, Hepatocellular; Humans; Liver Neoplasms; Male; Organotechnetium Compounds; Pyrrolidines; Radionuclide Imaging; Tetracycline; Tetracyclines

Topics: Adenocarcinoma; Biliary Fistula; Female; Humans; Liver Neoplasms; Middle Aged; Postoperative Complications; Sclerosing Solutions; Sigmoid Neoplasms; Tetracycline

The effects of numerous drugs and drug metabolites on the liver cleansing system are reviewed. Topics covered include metabolite-related drug toxicity, drug hypersensitivity, chronic liver disease, and hepatic tumors and oral contraceptives (OCs). Currently available evidence implicating OCs in the pathogenesis of hepatic tumors is large but inconclusive. However, both benign and malignant hepatic tumors have been encountered in women taking OCs, inviting further speculation that in some patients malignant transformation may occur. In a study covering 1937-1976, only 400 cases of hepatic cell adenoma and focal nodular hyperplasia were reported in patients, not taking OCs, but more than 200 cases have been reported in women taking OCs in the past 3 years. This may, however, present a biased view in favor of reporting contraceptive-associated tumors. An 8-year prospective study in Great Britain has so far not revealed tumors either in women receiving these agents or in any of their control groups. Studies with animals have been inconclusive so far; norethynodrel with mestranol (Enovid) failed to increase the incidence of hepatic tumors in susceptible mice, though a number of C17 alpha alkyl derivatives of testosterone will increase the expected frequency of hepatic tumors in susceptible mice. The reported regression of benign hepatic tumors on withdrawal of therapy and recurrences of tumors after primary resection in patients who continued to use such agents support the etiological role of OCs. However, more accurate statistics, which can only be obtained by large regional or national registries of hepatic tumors, are essential before this problem can be adequately evaluated.

Topics: Acetaminophen; Adult; Animals; Chemical and Drug Induced Liver Injury; Chronic Disease; Contraceptives, Oral; Drug Hypersensitivity; Female; Halothane; Humans; Isoniazid; Liver; Liver Neoplasms; Methotrexate; Pharmaceutical Preparations; Tetracycline

Iodine-131-tetracycline (131I-TET) was prepared by allowing tetracycline hydrochloride to react with radioiodide in acidic methanol (labeling efficiency greater than 85%). This preparation was found to be stable at--4 degrees C for at least 72 hr. Some minimal in vivo breakdown did occur. The 131I-TET, 67Ga, and several 99mTc compounds were studied in a rat hepatoma model. The incorporation of the radiopharmaceuticals into the tumor occurred rapidly, with peak levels at 0.5 and 24 hr after injection for 131I-TET and 67Ga, respectively. The clearnace of the radiopharmaceutical from nonviable tumor was slower than for viable tumor, and by 72 hr after injection the greatest concentration of radioactivity appeared in the nonviable fraction. All normal tissues showed faster clearance than did tumor tissue, regardless of viability. Decreasing the quantity of 131I-TET injected increased the percent of uptake in the nonviable tumor tissue but had no effect on the viable tumor uptake. Of the 99mTc compounds studied, the phosphates developed the highest tumor-to-background ratios. Unfortunately these ratios were not as high as those achieved for 67Ga or 131I-TET.

Topics: Animals; Disease Models, Animal; Gallium; Gallium Radioisotopes; Iodine Radioisotopes; Isotope Labeling; Liver Neoplasms; Neoplasm Transplantation; Neoplasms; Neoplasms, Experimental; Phosphates; Potassium Iodide; Radionuclide Imaging; Rats; Sugar Acids; Technetium; Tetracycline

A study was made of the efficacy of trypan blue, acridine orange, tetracycline and oxytetracycline for detection of tumour cells injected into the blood stream of rats. The cells were identified in the mesenteric microvessels by intravital microscopy. Fluorescence of fluorochromized cells was observed in the blue-violet (lambda max = 400 nm) and ultra-violet (lambda max = 365 nm) irradiation of the fluorescent lamp and in the laser irradiation (lambda = 337 nm). The cells stained with acridine orange had a higher fluorescence intensity and a more distinct structure than those labelled with tetracyclines. Identification of cells with trypan blue was more difficult. The fluorescent method of determination is rather simple and permits to indentify tumour cells directly in the blood stream.

Topics: Acridines; Animals; Carcinoma, Hepatocellular; Liver Neoplasms; Microscopy, Fluorescence; Neoplasms, Experimental; Neoplastic Cells, Circulating; Oxytetracycline; Rats; Staining and Labeling; Tetracycline; Trypan Blue

Topics: Adenocarcinoma; Animals; Carcinoma, Bronchogenic; Carcinoma, Hepatocellular; Gallium; Glioblastoma; Hodgkin Disease; Humans; L-Lactate Dehydrogenase; Liver Neoplasms; Lung Neoplasms; Mediastinal Neoplasms; Methane; Mice; Muscular Diseases; Neoplasm Metastasis; Neoplasm Transplantation; Neoplasms, Experimental; Nitrosourea Compounds; Osteosarcoma; Rabbits; Radioisotopes; Radionuclide Imaging; Rats; Sarcoma; Sarcoma, Experimental; Technetium; Tetracycline; Transplantation, Homologous

Topics: Animals; Carcinoma, Hepatocellular; Drug Stability; Gastric Mucosa; Kidney; Liver; Liver Neoplasms; Neoplasm Transplantation; Neoplasms, Experimental; Radionuclide Imaging; Rats; Rats, Inbred BUF; Technetium; Tetracycline

Topics: Aged; Bile; Carcinoma, Hepatocellular; Cholelithiasis; Humans; Infusions, Parenteral; Liver Neoplasms; Male; Middle Aged; Minocycline; Postoperative Complications; Tetracycline; Time Factors

Topics: Acridines; Animals; Carcinoma, Ehrlich Tumor; Carcinoma, Hepatocellular; Liver Neoplasms; Mice; Microscopy, Fluorescence; Neoplasms, Experimental; Radiation Effects; Rats; Tetracycline

Topics: Adult; Animals; Diagnosis, Differential; Echinococcosis, Hepatic; Echinococcus; Female; Hepatectomy; Humans; Laparotomy; Liver Neoplasms; Tetracycline; Thymol; Zoonoses

Topics: Animals; Fluorescence; Liver Neoplasms; Neoplasms, Experimental; Rats; Tetracycline

Topics: Adenoma; Aminosalicylic Acid; Aminosalicylic Acids; Animals; Anti-Bacterial Agents; Antineoplastic Agents; Antitubercular Agents; Carcinoma, Ehrlich Tumor; Carcinoma, Hepatocellular; Chloramphenicol; Isonicotinic Acids; Leukemia; Leukemia, Experimental; Liver Neoplasms; Neoplasms; Neoplasms, Experimental; Oleandomycin; Pharmacology; Pyrazinamide; Research; Rolitetracycline; Sarcoma; Sarcoma, Experimental; Sarcoma, Yoshida; Streptomycin; Tetracycline