Antineoplastic Agents ; therapeutic use ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; Humans ; Interferon-alpha ; therapeutic use ; Liver Neoplasms ; drug therapy ; pathology ; Neoplasm Recurrence, Local ; drug therapy ; prevention & control

OBJECTIVETo establish human hepatocellular carcinoma multidrug-resistance cell line (HepG2/ADM) and to determine the effect of low-frequency pulse ultrasound (US) on MDR cells and investigate its mechanism.

METHODSUsing gradual increase of adriamycin (ADM) concentrations in culture, an adriamycin-resistant human hepatocellular carcinoma cell sub line (HepG2/ADM) was established in vitro. HepG2/ADM cells were cultured in vitro and randomly divided into 4 groups: the control group (HepG2/ADM only), the group ADM by 1.0 mug/ml adriamycin for 1 h, the group US by low-frequency pulse ultrasound for 10 min, and the group US by low-frequency pulse ultrasound and treated with adriamycin simultaneously at same time. A sonication at a frequency of 0.8 MHz, was delivered with an intensity level of 0.5W/cm2, with continuous exposure of 10 min was applied. The ability of US to induce the apoptosis of MDR was evaluated by analyses of fluorescence microscopy, DNA fragmentation assay and flow cytometry assay.

RESULTSHepG2/Adm was resistant to many anti-tumor agents, and its IC50 of ADM was 26 times higher than that of parent cell line HepG2. Significant over expressions of P-gp, MRP, LRP and GSTs were detected. HepG2/ADM cells radiated by US had the typical characteristics of apoptosis. Compared with the control group (HepG2/ADM, 3.47%); the apoptosis rates were higher in US (12.23%). The therapeutic alliance of US with ADM for MDR cells, have a significant change in the ratio of apoptosis (18.81%, t=1.46 to 5.36, P<0.01).

CONCLUSIONHepG2/ADM could have the biological characteristics of human multidrug-resistance cell line. The US sonication of 0.8 MHz could induce apoptosis of HepG2/ADM cell in vitro, and could act synergistically with Adriamycin.

Apoptosis ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; Cell Line, Tumor ; DNA Fragmentation ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Flow Cytometry ; Humans ; Liver Neoplasms ; drug therapy ; pathology ; Ultrasonics

Antineoplastic Agents ; pharmacology ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; Cell Division ; drug effects ; Cisplatin ; pharmacology ; Humans ; Inhibitory Concentration 50 ; Liver Neoplasms ; drug therapy ; pathology ; Tumor Cells, Cultured ; drug effects

OBJECTIVETo investigate the inhibition effect of Salvia chinensis extraction (SJC) on growth of H22 bearing tumor, and to analyze the mechanism about the anti-tumor effect.

METHODWith hepatoma H22 bearing mice, ICR mice were inoculated with H22 cells by subcutaneous injection. On day 2 after inoculation, the ICR mice were randomize into 5 groups, they were the control group, the high, middle, low dosages of SJC, and the positive-control group. Administrated 10 days, the inhibition rate of tumor in the treatment groups were analyzed at 11th day. Meanwhile, Immunostaining with antibodies against CD105 and VEGF were used to investigate the tumor-related angiogenesis. In addition, the effect of SJC on angiogenesis of tumor were investigated on chick embry chorioallantoic membrane (CAM) inoculated H22 cells.

RESULTIn contrast to model group, the inhibition rate of the high, middle, and low dosages of SJC were 26.44% (P < 0.05), 42.28% (P < 0.01) and 32.59% (P < 0.05), respectively, and SJC could significantly reduced the express of VEGF and microvessel density (MVD) (P < 0.01). Injection with 6.25 g x L(-1) doses of SJC could significantly inhibited the angiogenesis of tumor, the inhibition rate of tumor-related angiogenesis was 50.67% (P < 0.01).

CONCLUSIONSJC showed anticancer effect, and maybe it is related to down-regulation VEGF and reducing MVD, then inhibiting the tumor-related angiogenesis.

Animals ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; pathology ; Cell Line, Tumor ; Male ; Mice ; Mice, Inbred ICR ; Neovascularization, Pathologic ; drug therapy ; metabolism ; pathology ; Plant Extracts ; therapeutic use ; Salvia ; chemistry

OBJECTIVETo screen out the HAb18G/CD147 binding peptides and find out an antagonist against hepatoma invasion.

METHODSHAb18G/CD147 was purified by affinity chromatographic method and the antigen binding peptides acquired by bio-panning a phage-displayed 12-peptide library. After obtaining the sequence of the selected phage-displayed peptides, all the 9 peptides were synthesized by solid-phase method and identified by mass spectrograph. The peptides' anti-metastatic function was tested by Boyden Chamber assay.

RESULTSThe purified HAb18G/CD147, identified by Western blot (molecular weight about 65 kd) could be used to bio-pan the phage-displayed peptide library. After 3 rounds of bio-panning, 9 positive phage clones were selected and sequenced. The synthesized peptides had uneven inhibitory activities and three of them were able to markedly inhibit the hepatoma cell invasion (P < 0.01). The most effective peptide decreased by 90.1% of hepatoma cells migrating through the Boyden Chamber membrane as compared with the control.

CONCLUSIONBio-panning the phage-displayed peptide library can be used successfully to screen out the antigen binding peptides. Hepatoma metastatic potential can be inhibited by peptide antagonist which could be a good foundation of developing peptide therapeutic agent against hepatoma metastasis.

Animals ; Antibodies, Monoclonal ; therapeutic use ; Basigin ; metabolism ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; Humans ; Liver Neoplasms ; drug therapy ; pathology ; Mice ; Neoplasm Invasiveness ; Peptide Library ; Peptides ; therapeutic use

The purpose of this study was to investigate the effect of Huaier on autophagy of human hepatoma SK-HEP-1 cells and the effect of autophagy on the proliferation of SK-HEP-1 cells. CCK-8 assay was used to evaluate the effect of Huaier on the proliferation of SK-HEP-1 cells under different concentrations and different times. Acridine orange staining was used to measure the effect of Huaier on the autolysosome formation in SK-HEP-1 cells. Immunofluorescence assay was applied to examine the effect of Huaier on the expression and distribution of autophagy marker LC3 in SK-HEP-1 cells. In addition， LC3 expression was also checked by immunoblot analysis in the presence of Huaier. At last， the effects of Huaier in combination with autophagy inhibitor bafilomycin A1 on the proliferation of SK-HEP-1 cells was detected by CCK-8 assay. The results showed that Huaier aqueous extract significantly inhibited the proliferation of human hepatoma SK-HEP-1 cells in a dose- and time-dependent manner. Huaier aqueous extract dramatically promoted the formation of autolysosome in SK-HEP-1 cells. Moreover， Huaier markedly increased the number and intensity of intracellular LC3 fluorescent puncta and up-regulated LC3-Ⅱ expression. These data indicated that Huaier evidently activated autophagy of SK-HEP-1 cells. Additionally， autophagy inhibition significantly attenuated the sensitivity of SK-HEP-1 cells to Huaier treatment. Therefore， autophagy activation is involved in the inhibitory effects of Huaier on the proliferation of human hepatoma SK-HEP-1 cells.
Apoptosis ; Autophagy ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Complex Mixtures ; pharmacology ; Humans ; Up-Regulation

Antineoplastic Agents ; pharmacology ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; Cell Division ; drug effects ; Dose-Response Relationship, Drug ; Drug Synergism ; Fluorouracil ; pharmacology ; Humans ; Liver Neoplasms ; drug therapy ; pathology ; Tumor Cells, Cultured ; drug effects

INTRODUCTIONPatients with peritoneal metastases (PM) from hepatocellular carcinoma (HCC) often experience a rapid demise even after a complete removal of intrahepatic tumour. Localised PM may now be adequately controlled and managed with cytoreductive surgery (CRS).

TREATMENTThree patients underwent CRS for HCC PM.

OUTCOMEThe first patient survived 21 months from the time of CRS and is alive with the disease. The second patient died 4 months after CRS. The third patient survived 10 months since CRS and is also alive with the disease. Collectively, the survival of 24 patients with HCC PM extracted through a collective literature review who were treated with cytoreductive surgery had 1- and 2-year survival percentages of 83% and 71%, respectively.

CONCLUSIONCareful selection of patients with localised disease to the peritoneal cavity for CRS, taking into consideration the performance status, liver function and tumour biology may lead to a successful outcome in patients with HCC PM.

Carcinoma, Hepatocellular ; drug therapy ; pathology ; surgery ; Fatal Outcome ; Female ; Humans ; Liver Neoplasms ; drug therapy ; pathology ; surgery ; Male ; Middle Aged ; Peritoneal Neoplasms ; drug therapy ; secondary ; surgery ; Peritoneum ; pathology ; Young Adult

OBJECTIVE: To investigate the antitumor activity of decoction and study its liver and kidney toxicity and its effect on the immune system in a tumor-bearing mouse model. METHODS: Hepatoma H22 tumor-bearing mouse models were randomized into model group, cyclophosphamide (CTX) group, and low-, moderate-, and high-dose decoction groups (JW-L, JW-M, and JW-H groups, respectively). The antitumor activity of decoction was assessed by calculating the tumor inhibition rate and pathological observation of the tumor tissues. Immunohistochemistry was used to detect the expressions of Bax, Bcl-2, Bax/Bcl-2 and caspase-3 in the tumors. The liver and kidney toxicity of decoction was analyzed by evaluating the biochemical indicators of liver and kidney functions. The immune function of the tumor-bearing mice were assessed by calculating the immune organ index, testing peripheral blood routines, and detection of serum IL-2 and TNF-α levels using enzyme-linked immunosorbent assay. RESULTS: Compared with that in the model group, the tumor mass in CTX, JW-M and JW-H groups were all significantly reduced ( < 0.05) with cell rupture and necrosis in the tumors. Immunohistochemistry revealed obviously up-regulated expressions of Bax and caspase-3 and down- regulated expression of Bcl-2 protein with an increased Bax/Bcl-2 ratio in CTX, JW-M and JW-H groups. Treatment with decoction significantly reduced Cr, BUN, AST and ALT levels, improved the immune organ index, increased peripheral blood leukocytes, erythrocytes and hemoglobin levels, and up-regulated the levels of TNF-α and IL-2 in the tumor-bearing mice. These changes were especially significant in JW-H group when compared with the parameters in the model group ( < 0.01). CONCLUSIONS decoction has a strong anti-tumor activity and can improve the liver and kidney functions of tumor-bearing mice. Its anti-tumor effect may be attributed to the up-regulation of Bax, caspase-3, TNF-α and IL-2 levels and the down-regulation of Bcl-2 expression as well as the enhancement of the non-specific immune function.
Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Carcinoma, Hepatocellular ; drug therapy ; immunology ; metabolism ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Kidney ; drug effects ; Liver ; drug effects ; pathology ; Liver Neoplasms ; drug therapy ; immunology ; metabolism ; pathology ; Mice ; Necrosis ; Neoplasm Proteins ; metabolism ; Random Allocation ; Up-Regulation

This study was to examine the mechanism of oleanolic acid (OA) induces G2/M phase arrest and apoptosis in human hepatocellular carcinoma Bel-7402 cells. MTT and trypan blue exclusion test assay were adopted to detect the proliferate status of cells treated with OA. We assayed the cell cycle by flow cytometry using PI staining. Apoptosis was determined by Annexin V-FITC staining and PI labeling. The expressions of cycle related proteins and apoptotic related proteins were determined by Western blot analysis. OA strongly inhibited human hepatoma cells proliferation. When Bel-7402 cells were pretreated with OA for 24 h, OA induced apoptosis and G₂/M phase cell cycle arrest in a concentration-dependent manner. Analysis of the cell cycle regulatory proteins demonstrated that OA decreased the protein levels of cyclin B1, but increased the protein levels of p-Cdk1 (Tyr15) and p-Cdc25C (Ser 216). Moreover, OA modulated the phosphorylation of protein kinases Chk1 and p2l. Western blotting assay also showed significant decrease of Bcl-2 protein expression and increase of Bax protein expression, the cytosol Cyt c level, cleaved-caspase-9 and cleaved-caspase-3 activity. These data suggest that OA produces anti-tumor effect via induction of G₂/M cell cycle arrest and apoptosis.
Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; Cell Line, Tumor ; G2 Phase Cell Cycle Checkpoints ; drug effects ; Humans ; Liver Neoplasms ; drug therapy ; pathology ; M Phase Cell Cycle Checkpoints ; drug effects ; Oleanolic Acid ; pharmacology