Promyelocytic leukemia protein (PML) is a major component of PML nuclear bodies (PML NBs). Fusion of promyelocytic leukemia gene (PML) with retinoic acid receptor alpha gene with the t (15;17) translocation causes disassembly of PML NBs, leading to development of acute promyelocytic leukemia. In contrast, PML overexpression as well as different morphological changes of PML NBs were described in a few solid tumors. In this study, the expression of PML through the multistep hepatocarcinogenesis was analyzed in 95 cases of human hepatocellular carcinomas (HCCs) for comparison along with dysplastic nodules (DNs) and background liver cirrhosis (LC) or chronic hepatitis by immunohistochemistry and immunoblot. In addition, cases of HCCs were further evaluated according to their histologic grade and etiology. The amount of PML as well as the num-ber and size of PML NBs increased gradually through the progression from LC, DNs to HCCs. The overexpression of PML in HCCs was much more closely associated with HBV infection than HCV infection or alcoholic liver disease. The PML expression, however, was not correlated with histologic grade of HCCs. These results suggest that PML is involved in the early stage of multistep hepatocarcinogenesis, and HBV infection may be associated with the overexpression of PML and the morphological alteration of PML NBs.
Carcinoma, Hepatocellular/*chemistry/ultrastructure ; Cell Nucleus/*chemistry ; Human ; Liver/chemistry ; Liver Neoplasms/*chemistry/ultrastructure ; Neoplasm Proteins/*analysis ; Precancerous Conditions/*chemistry/ultrastructure ; Transcription Factors/*analysis ; Tumor Cells, Cultured

Histochemical, immunohistochemical and ultrastructural studies were performed on cases of hepatocellular carcinoma (HCC) with pale bodies (PB). HCC containing PBs was observed in 3 (5.5%) of 55 consecutively resected HCC cases. Histologically, a large number of hepatocytes displayed pale or eosinophilic staining of the cytoplasm, resulting in ground-glass appearance. They were aggregated in nodular pattern, or diffusely intermixed with other malignant hepatocytes. PBs were negative for periodic-acid Schiff and Masson's trichrome staining. The inclusions showed a strong positive reaction for fibrinogen and some of them were weakly positive for albumin but negative for hepatitis B surface antigen, hepatitis B core antigen, alpha-fetoprotein and alpha-1-antitrypsin. Ultrastructurally, PBs were membrane-bound and contained granular materials of moderate electron density, and were closely related to dilated rough endoplasmic reticulum. These findings support that PBs are secretory fibrinogen accumulated in cystic ER and that such intracellular accumulation possibly reflects a defective transport of fibrinogen.
Albumins/analysis ; Carcinoma, Hepatocellular/pathology* ; Cytoplasm/ultrastructure ; Cytoplasm/pathology ; Cytoplasm/chemistry ; Endoplasmic Reticulum, Rough/ultrastructure ; Endoplasmic Reticulum, Rough/pathology ; Endoplasmic Reticulum, Rough/chemistry ; Fibrinogen/analysis ; Human ; Inclusion Bodies/ultrastructure ; Inclusion Bodies/pathology* ; Inclusion Bodies/chemistry ; Liver Neoplasms/pathology* ; Male ; Microscopy, Electron ; Middle Age ; Periodic Acid-Schiff Reaction

OBJECTIVETo investigate the anti-cancer activities and the possible mechanism of Chinese herb Dioscrea bulbifera.

METHODThe herb was extracted sequentially with petroleum ether, ethanol and water. The anticancer screen were carried out in vivo with HepA in mice.

RESULTThe inhibitory effects on the formation of ascites volume and HepA cell viability in ascites were found in those extracted fractions except water fraction, the petroleum ether fraction being the strongest. Life span of mice bearing HepA ascites was prolonged after exposed to 100 mg x kg(-1) petroleum ether fraction and shortened after exposed to water fraction significantly. Besides, abnormal microstructure on HepA cells surface was found and it was supposed to be potential effect against viability of HepA which was convinced with the regeneration of HepA cells from ascites in mice exposed to petroleum ether fraction.

CONCLUSIONAnticancer active compounds are mainly extracted by petroleum ether from hydrophobic constituents of Dioscrea bulbifera and the anticancer effects were related to direct toxicity on tumor cell.

Alkanes ; Animals ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Ascites ; pathology ; Carcinoma, Hepatocellular ; pathology ; ultrastructure ; Cell Line, Tumor ; Cell Survival ; drug effects ; Dioscorea ; chemistry ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Ethanol ; Liver Neoplasms ; pathology ; ultrastructure ; Male ; Mice ; Mice, Inbred ICR ; Neoplasm Transplantation ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry

OBJECTIVETo investigate the anti-cancer effects of crude extract from Melia toosendan Sieb. et Zucc and its possible molecular mechanisms in vitro and in vivo.

METHODSTransonic alcohol-chloroform extraction method was used to extract toosendanin from the bark of Melia toosendan Sieb. et Zucc, and the content of toosendanin in the crude extract was measured by high performance liquid chromatography (HPLC). Anti-cancer effects of crude extract from Melia toosendan Sieb. et Zucc were investigated in in vivo and in vitro studies. In the in vitro experiment, human hepatocellular carcinoma cell lines SMMC-7721 and Hep3B were co-incubated with toosendanin crude extract of different concentrations, respectively. In the in vivo experiment, BALB/c mice were subcutaneously inoculated with mouse hepatocellular carcinoma H22 cells and treated with crude extract.

RESULTSHPLC revealed the content of toosendanin was about 15%. Crude extract from Melia toosendan Sieb. et Zucc inhibited cancer cells growth in a dose- and time-dependent manner. The 50% inhibitory concentration (IC50, 72 h) was 0.6 mg/L for SMMC-7721 cells and 0.8 mg/L for Hep3B cells. Both high-dose [0.69 mg/(kg d)] and low-dose [0.138 mg/(kg d)] crude extract could markedly suppress cancer growth, and the inhibition rate was greater than 50%. Hematoxylin and eosin staining showed necrotic area in cancers and transmission electron microscopy displayed necrotic and apoptotic cancer cells with apoptotic bodies. Immunohistochemistry showed that the expression of Bax and Fas increased and the expression of Bcl-2 reduced.

CONCLUSIONSToosendanin extract has potent anti-cancer effects via suppressing proliferation and inducing apoptosis of cancer cells in vivo and in vitro. The mechanism of apoptosis involves in mitochondrial pathway and death receptor pathway.

Animals ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; ultrastructure ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; therapeutic use ; Female ; Immunohistochemistry ; Liver Neoplasms ; drug therapy ; pathology ; ultrastructure ; Male ; Melia ; chemistry ; Mice, Inbred BALB C ; Mitochondria ; drug effects ; metabolism ; Neoplasm Transplantation ; Plant Extracts ; therapeutic use ; Reference Standards ; bcl-2-Associated X Protein ; metabolism ; fas Receptor ; metabolism

OBJECTIVETo observe the effects of Scutellaria barbata extract (ESB) on human hepatoma cell line Hep-G2 proliferation in vitro and explore the mechanism.

METHODSThe inhibitory effect of ESB on Hep-G2 proliferation was estimated by MTT assay, and the morphological changes of the cells were observed under optical and electron microscopes. Distribution of cell cycle, cell apoptosis and the protein expressions of apoptosis-associated genes as bcl-2, bax and fas were analyzed using flow cytometry.

RESULTSESB inhibited the proliferation of Hep-G2 cells in a time- and dose-dependent manner. ESB treatment for 72 h resulted in changes of early apoptotic morphology of the cells as observed under optical and the transmission electron microscopes and increased cell apoptosis. Cell cycle analysis revealed decreased S-phase and increased G0/G1-phase cells. Fas expression was significantly up-regulated in response to ESB treatment whereas Bcl-2 and Bax expressions underwent no significant changes.

CONCLUSIONESB can inhibit Hep-G2 cell proliferation, induce cell cycle block, and increase cell apoptosis, which may relate to the activation of FNFR superfamily.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; pathology ; physiopathology ; ultrastructure ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Flow Cytometry ; Humans ; Liver Neoplasms ; pathology ; physiopathology ; ultrastructure ; Microscopy, Electron, Transmission ; Plant Extracts ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Scutellaria ; chemistry ; bcl-2-Associated X Protein ; analysis ; fas Receptor ; analysis