With the aim of establishing bio-indices for the development of multistep hepatotumorigenesis, rats were fed water containing 0.01% diethylnitrosamine (DEN) ad libitum for 13 weeks. This treatment with DEN only made it possible to induce hepatic tumors in 100%. After the DEN administration, several clinical symptoms were observed including minor behavioral changes, brittleness of hair and a decrease in water and food intake. The concentration of total serum protein and albumin in all treated groups was significantly lower than in non-treated controls (p<0.05). Increase of specific enzyme (AST, ALT and GGT) activity (p<0.05), variable tumor size and hepatomegaly of the liver was observed in all rats treated with DEN for 10 weeks. Both hepatocellular carcinoma and cholangiocarcinoma were found in the same livers at the same time, and were prominently developed after 12 weeks. In case of carcinoma, some of the livers showed more or less advanced states over the 12-15 weeks period. In the present study, hepatocellular carcinoma was developed by treating DEN in only the drinking water, without any other carcinogens or without partial hepatectomy. These results indicate that DEN is a new carcinogen that acts directly on it the liver, moreover, it might be very useful for investigating hepatotumorigenesis.
Alanine Transaminase/blood ; Animals ; Aspartate Aminotransferases/blood ; Biological Markers/blood ; Carcinogens ; *Cell Transformation, Neoplastic ; Diethylnitrosamine/toxicity ; Liver/drug effects/*pathology ; Liver Neoplasms/blood/chemically induced/*pathology ; Liver Neoplasms, Experimental/blood/*pathology ; Male ; Rats ; Rats, Sprague-Dawley ; gamma-Glutamyltransferase/blood

Animals ; Antineoplastic Agents ; administration & dosage ; Electroporation ; methods ; Interleukin-2 ; blood ; Interleukin-6 ; blood ; Liver Neoplasms, Experimental ; immunology ; mortality ; pathology ; therapy ; Male ; Neoplasm Transplantation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the dynamic expressions of TGF-beta 1 and TGF-beta 1 mRNA at different stages of hepatocellular carcinoma (HCC) development and their use in clinical diagnosis.

METHODSHepatoma models were developed with 2-FAA using male Sprague-Dawley (SD) rats. Morphological changes of the rat liver histological preparations (H and E stained) were studied. The fragment of TGF-beta 1 gene obtained was amplified by nested RT-PCR. Dynamic change of TGF-beta 1 level was quantitatively analyzed by ELISA. The distribution of TGF-beta 1 in the cells and its gene expression were detected in human HCC tissues.

RESULTSThe progressive increases of hepatic TGF-beta 1 and TGF-beta 1 mRNA were observed in rat hepatocytes which progressed from granular degeneration, atypical hyperplasia and finally to HCC development induced by 2-FAA. The expression levels in HCC tissues were significantly higher than those in the normal and degenerative ones. TGF-beta 1 was shown in rat hepatocytes by immunohistochemistry. Plasma TGF-beta 1 was detected in 89.5% of all the patients with HCC, but it was detected in 93.3% of them who had an AFP less than 400 microg/L. TGF-beta 1 mRNA showed a stronger expression in HCC tissues. TGF-beta 1 mRNA was found in peripheral blood mononuclear cells from all HCC patients with extrahepatic metastasis.

CONCLUSIONTGF-beta 1 may participate in hepatocyte canceration. The overexpression of TGF-beta 1 and TGF-beta 1 mRNA could be useful markers for early diagnosis and predicting prognosis of HCC.

Adult ; Aged ; Animals ; Biomarkers, Tumor ; blood ; Carcinoma, Hepatocellular ; blood ; diagnosis ; pathology ; Female ; Humans ; Liver Neoplasms, Experimental ; blood ; diagnosis ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; prevention & control ; Prognosis ; RNA, Messenger ; blood ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; blood

OBJECTIVETo investigate the inhibitory effects and its mechanism of lyophilized powder of fresh Gekko Chinenis (GCLP) on H22 hepatocarcinoma growth and angiogenesis in vitro.

METHODSThe transplant tumor model of H22 hepatocarcinoma in mice was established. Thirty mice were randomly divided into three groups, the cisplatin group, the GCLP group and the control group, they were treated respectively with intraperitoneal injection of cisplatin 1 mg/g, oral administration of GCLP in a dose of 1.2 g/kg, and equal volume of saline, the medication was given for 20 times totally. The anti-tumor activity was evaluated by tumor tissue weighing, the cell apoptotic rate was detected by TUNEL method, the micro-vessel density in tumor tissue was determined by Weidner method, the protein expression of vascular endothelin growth factor (VEGF) and basic fibroblast growth factor (bFGF) were detected by S-P immunohistochemistry.

RESULTSGCLP could obviously inhibit the hepatocarcinoma growth, induce tumor cell apoptosis, and reduce micro-vessel density in tumor tissue through down-regulating VEGF and bFGF protein expression.

CONCLUSIONSGCLP can effectively inhibit the growth of H22 hepatocarcinoma and angiogenesis. Its mechanism might be related to the down-regulation of the protein expression of VEGF and bFGF.

Angiogenesis Inhibitors ; pharmacology ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Liver Neoplasms, Experimental ; blood supply ; pathology ; Lizards ; Materia Medica ; Mice ; Neovascularization, Pathologic ; Tumor Cells, Cultured

To investigate the influence of bear bile on rat hepatocarcinoma induced by diethylnitrosamine (DEN), a total of 40 rats were randomly divided into 4 groups: normal control group, model group, and two bear bile treatment groups. The rat liver cancer model was induced by breeding with water containing 100 mg x L(-1) DEN for 14 weeks. The rats of the bear bile groups received bear bile powder (200 or 400 mg x kg(-1)) orally 5 times per week for 18 weeks. The general condition and the body weight of rats were examined every day. After 18 weeks the activities of serum alanine transaminase (ALT), aspartate transaminase (AST) and total bilirubin (TBIL) were detected. Meanwhile, the pathological changes of liver tissues were observed after H&E staining. The expression of proliferative cell nuclear antigen (PCNA) and a-smooth muscle actin (alpha-SMA) in liver tissue were detected by immunohistochemical method. After 4 weeks the body weights of rats in normal group were significantly more than that in other groups (P < 0.05); and that in the two bile groups was significantly more than that in the model group. Compared with normal group, the level of serum glutamic-pyruvic transaminase and total bilirubin increased significantly in other groups; compared with model group, these two indexes decreased significantly in two bile groups. Hepatocellular carcinoma occurred in all rats except for normal group; there were classic cirrhosis and cancer in model group while there were mild cirrhosis and high differentiation in two bile groups. There were almost no expressions of PCNA and alpha-SMA in normal group while there were high expressions in model group; the two bile groups had some expressions but were inferior to the model group, and alpha-SMA reduced markedly. It indicated that bear bile restrained the development of liver cancer during DEN inducing rat hepatocarcinoma, which may be related to its depressing hepatic stellate cell activation and relieving hepatic lesion and cirrhosis.
Actins ; metabolism ; Alanine Transaminase ; blood ; Animals ; Antineoplastic Agents ; pharmacology ; Aspartate Aminotransferases ; blood ; Bile ; chemistry ; Bilirubin ; blood ; Body Weight ; drug effects ; Carcinoma, Hepatocellular ; blood ; chemically induced ; pathology ; Diethylnitrosamine ; Liver ; metabolism ; pathology ; Liver Cirrhosis ; chemically induced ; pathology ; Liver Neoplasms, Experimental ; blood ; chemically induced ; pathology ; Male ; Powders ; pharmacology ; Proliferating Cell Nuclear Antigen ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Ursidae

OBJECTIVETo observe the effect of traditional Chinese herbs Fuzheng Yiliu Granule (FYG)-contained serum from tumor bearing mice on apoptotic rate, free radicals content and mitochondrial membrane potential of hepatoma cell lines H22 in vitro.

METHODSThe effect of FYG drug-serum on apoptosis of hepatoma cell line H22 was determined using flow cytometry. The changes of DNA RNA, free radicals and mitochondrial membrane potential (delta psi m) in H22 cell were detected through laser scanning confocal microscope.

RESULTSFYG-contained serum can induce the apoptosis of H22 cell, enhance the free radicals content, and reduce the content of DNA RNA and delta psi m of H22 cell in vitro.

CONCLUSIONThe apoptosis of hepatoma cell line H22 induced by FYG is probably correlated to the change of free radicals content and delta psi m.

Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Drugs, Chinese Herbal ; pharmacology ; Flow Cytometry ; Free Radicals ; metabolism ; Liver Neoplasms, Experimental ; blood ; metabolism ; pathology ; Membrane Potential, Mitochondrial ; drug effects ; Mice ; Microscopy, Confocal ; Serum

To study the influence of recombinant endostatin on angiogenesis and tumor growth of mice H22 hepatoma, tumor models were constructed by injecting H22 hepatoma cells into the leg muscle of mice. Recombinant endostatin was produced by gene engineering in E. coli. The recombinant protein was injected subcutaneously to treat transplanted hepatoma faraway. The weight of tumors was measured, and the changes of necrosis of tumor cells and vessel density were observed by immunohistochemistry. The results suggested that the growth of hepatoma models transplanted in the muscle of legs was suppressed by recombinant endostatin. The density of vascularity was decreased, but the necrosis of tumor cells increased. The inhibitory effect of recombinant endostatin on angiogenesis and tumor growth of hepatoma was not affected after chemotherapy.
Angiogenesis Inhibitors ; pharmacology ; Animals ; Antineoplastic Agents ; pharmacology ; Endostatins ; biosynthesis ; genetics ; pharmacology ; Escherichia coli ; genetics ; Liver Neoplasms, Experimental ; blood supply ; pathology ; Male ; Mice ; Neoplasm Transplantation ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology

OBJECTIVETo study the efficacy and explore the mechanism of the anti-tumor immunity elicited by heat shock protein 70-peptide complexes (HSP70-PC) derived from tumor cells.

METHODSCells culture, flow cytometric analysis, affinity chromatography for protein purification, SDS-PAGE, Western-blotting and animal experiment were used.

RESULTSHSP70-PC immunization rendered protective effect to both naive tumorl-bearing mice. All of the naive mice obtained complete resistance to Hcaf cell attack; 40% of the tumor-bearing mice survived for over 90 days, whereas the mice of control group died within 2 weeks (P < 0.01). CD8+ subset of T lymphocytes in the peripheral blood of immunized mice increased by 12%.

CONCLUSIONHSP70-PC induces anti-tumor immunity via activation of cytotoxic T lymphocytes (CTLs), and it possesses strong tumor vaccine effect. Our research adds more evidence to support the clinical use of HSP70-PC to fight human cancers.

Animals ; CD8 Antigens ; blood ; Cell Membrane ; chemistry ; HSP70 Heat-Shock Proteins ; biosynthesis ; isolation & purification ; therapeutic use ; Liver Neoplasms, Experimental ; chemistry ; drug therapy ; immunology ; pathology ; Mice ; Neoplasm Transplantation ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Cells, Cultured

OBJECTIVETo investigate the changes of several protein markers in a metastatic colorectal carcinoma model by serum proteomic analysis.

METHODSThe pEGFP-N1 plasmid with enhanced expression of green fluorescence protein (EGFP) was transfected into human colon carcinoma cell line SW480 to obtain a stable SW480-EGFP cell line, the SW480-EGFP cells were then injected subcutaneously into nude mice. The harvested tumor cells were implanted orthotopically into the colon of the nude mice. Real-time tumor growth and metastasis formation were visualized by whole-body fluorescent imaging system. Serum samples at different metastatic stages were collected and differential proteomic profiles were investigated by two-dimensional gel electrophoresis (2DE) and matrix-assisted laser absorption/ionization time of flight mass spectrometry (MALDI-TOF MS).

RESULTSThe SW480- EGFP cells enabled to express EGFP stably. The rates of subcutaneous and orthotropic tumor formation were 100%. The metastasis rates to local lymph nodes, liver and lung were 100%, 40% and 30%, respectively. Furthermore, 5 differentially expressed proteins were analyzed by serum proteome technologies, including haptoglobin alpha chain, apolipoprotein E, apolipoprotein A-IV, Ig kappa chain V region chain L and transferrin.

CONCLUSIONSVisualized metastatic model of colorectal carcinoma was successfully established. Several differentially expressed serum proteins collected at different stages after the occurrence of metastasis were identified. These differentially expressed proteins may be candidate serum biomarkers for diagnosis and therapeutic evaluation of colorectal carcinoma metastasis.

Animals ; Apolipoproteins A ; blood ; Apolipoproteins E ; blood ; Biomarkers, Tumor ; blood ; Blood Proteins ; analysis ; Cell Line, Tumor ; Colorectal Neoplasms ; blood ; genetics ; pathology ; Electrophoresis, Gel, Two-Dimensional ; Green Fluorescent Proteins ; genetics ; metabolism ; Haptoglobins ; analysis ; Humans ; Immunoglobulin kappa-Chains ; blood ; Liver Neoplasms ; secondary ; Lung Neoplasms ; secondary ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neoplasms, Experimental ; blood ; genetics ; pathology ; Proteomics ; methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Transfection ; Transferrin ; analysis ; Transplantation, Heterologous

OBJECTIVETo study the anti-tumor activity of extractive from Celastrus orbiculatus in vivo.

METHODMice bearing transplanted tumor S180 and Heps were used to study the effects of acetoacetate and n-butanol extractive from C. orbiculatus. The changes in serum contents of SOD and malondialdehyde (MDA) content were assayed.

RESULTAcetoacetate and n-butanol extractive from C. orbiculatus significantly inhibited the growth of S180 and Heps tumor in mice. SOD content was obviously increased, MDA content obviously decreased in the serum after extractive treatment.

CONCLUSIONAcetoacetate and n-butanol extractive from C. orbiculatus have anti-tumor effects and anti-oxidative capacity.

Animals ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Antioxidants ; isolation & purification ; pharmacology ; Celastrus ; chemistry ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Liver Neoplasms, Experimental ; blood ; pathology ; Male ; Malondialdehyde ; blood ; Mice ; Mice, Inbred ICR ; Neoplasm Transplantation ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Sarcoma 180 ; blood ; pathology ; Superoxide Dismutase ; blood