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

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

OBJECTIVETo study the effect of transcatheter arterial chemoembolization (TACE) combined with beta-ultrasound guided portal vein embolization (PVE) through fine-needle liver puncture for hepatocellular carcinoma.

METHODS209 patients with primary hepatocellular carcinoma were divided into TACE group (104 patients) and TACE + PVE group (105 patients).

RESULTSThe response rates (CR + PR) were 37.5% in TACE group and 57.2% in TACE + PVE group (P < 0.01). Tumor thrombi became lessened or resolved in the portal vein with incidences of 22.2% in TACE group and 68.8% in TACE + PVE group (P < 0.01). The 1-, 2- and 3-year survival rates were 65.1%, 36.3% and 20.5% in TACE group and 95.6%, 59.6% and 39.1% in TACE + PVE group (P < 0.05).

CONCLUSIONThe effect of TACE combined with PVE is much more effective than TACE alone for hepatocellular carcinoma. Beta-ultrasound guided portal vein embolization through fine-needle liver puncture is effective, easy, safe and should be widely practiced.

Adult ; Aged ; Arteries ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; pathology ; therapy ; Catheterization ; Chemoembolization, Therapeutic ; adverse effects ; methods ; Female ; Follow-Up Studies ; Humans ; Liver Neoplasms ; drug therapy ; metabolism ; pathology ; therapy ; Male ; Middle Aged ; Portal Vein ; Treatment Outcome ; alpha-Fetoproteins ; metabolism

OBJECTIVETo investigate the effects of peptide YY (PYY) on subcutaneous transplantation tumor of human hepatoma in nude mice and preliminarily explore the mechanisms.

METHODSHepG2 human hepatic carcinoma cells were injected into nude mice subcutaneously, and the resultant tumor were taken and prepared into small tissue blocks. The tissue blocks were implanted subcutaneously into nude mice to establish mouse models bearing human hepatoma. Thirty-two such mouse models were assigned equally into 4 groups to receive subcutaneous PYY injection at a high or low dose, intraperitoneal injection of floxuridine (positive control group), or subcutaneous normal saline injection (negative control group). The general condition of the tumor-bearing mice and the growth of the tumors were observed.

RESULTSCompared with the negative control group, the high- and low-dose PYY groups showed reduced gross tumor volume, lowered serum AFP, tumor weight, and cAMP content in the tumor tissue (P<0.05).

CONCLUSIONPYY can inhibit the growth of subcutaneous hepatoma in nude mice, which might be associated with the reduction of cAMP content in the tumors following PYY treatment.

Animals ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; Cell Line, Tumor ; Cyclic AMP ; metabolism ; Humans ; Liver Neoplasms, Experimental ; drug therapy ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Peptide YY ; therapeutic use ; Tumor Burden ; drug effects ; Xenograft Model Antitumor Assays

We surveyed the literatures domestic and abroad, and summarized traditional Chinese medicine (single or complex prescription) with anti-hepatoma effects by adjusting immune function. Traditional Chinese medicine showed great advantages in improving the immune system function of the organism in various ways, so they could prohibit the generation and development of tumor, lessen the damage caused by chemotherapeutics, increase the sensitivity of chemotherapeutics, strengthen immune surveillance to tumor cells, elevate living quatity of patients and prolong their living time. It is very promising to exploit much more effective anti-tumor drugs from TCM.
Animals ; Antineoplastic Agents, Phytogenic ; therapeutic use ; Carcinoma, Hepatocellular ; drug therapy ; immunology ; pathology ; Drugs, Chinese Herbal ; isolation & purification ; therapeutic use ; Humans ; Killer Cells, Natural ; drug effects ; immunology ; Liver Neoplasms ; drug therapy ; immunology ; metabolism ; Lymphocyte Activation ; drug effects ; Phytotherapy ; Plants, Medicinal ; chemistry

OBJECTIVESTo investigate the apoptosis induced by antisense oligodeoxynucleotide (ASODN) against survivin and the mechanisms after the hepatocellular carcinoma SMMC-7721 cells transfected with the ASODN.

METHODSThe ASODN was transfected into SMMC-7721 cells mediated by liposomal reagent. The changes of cell cycle and apoptotic rate were detected by flow cytometry. The changes of cell skeleton was observed through confocal microscope. The activity of p38MAPK and caspase-3 were detected by immuno-precipitation and kinase activity assess methods, respectively.

RESULTSThere were control, sense control, 400, 600, 800, and 1 000 ng/ml ASODN groups (I - VI). The apoptotic rats were 0.70%, 0.76%, 2.43%, 7.82%, 23.11%, and 31.35% in groups I - VI, respectively, which in the ASODN-transfected groups were higher than that in the control group (tor=20.9, P<0.01). The activity of p38MAPK increased significantly, when the ASODN was transfected at the concentration of 600 ng/ml or more, so did the caspase-3 activity (the p38MAPK and caspase-3 activity in groups I - VI were 7.03, 7.07, 13.47, 16.37, 43.97, 47.87 and 0.015+/-0.010, 0.014+/-0.002, 0.026+/-0.003, 0.042+/-0.001, 0.093+/-0.001, 0.100+/-0.001, respectively).

CONCLUSIONSASODN targeting at survivin mRNA can induce G2/M stop, activate p38MAPK and caspase-3. The activated caspase-3 destroys the cell skeleton microfilament system, resulting in apoptosis.

Apoptosis ; Carcinoma, Hepatocellular ; drug therapy ; enzymology ; pathology ; Caspase 3 ; Caspases ; metabolism ; Cell Line, Tumor ; Humans ; Inhibitor of Apoptosis Proteins ; Liver Neoplasms ; drug therapy ; enzymology ; pathology ; Microtubule-Associated Proteins ; antagonists & inhibitors ; genetics ; Neoplasm Proteins ; Oligonucleotides, Antisense ; therapeutic use

Control of inflammation is widely accepted as an important strategy for cancer chemoprevention. Anti-inflammatory effects of bark extracts of elm tree (BEE) have been amply reported. Therefore, BEE may be a good candidate cancer chemopreventive agent. Considering the high incidence of hepatic cancer and limited therapeutic approaches for treating this disease, it is important to develop liver cancer-specific chemopreventive agents. To evaluate the chemopreventive potential of BEE, we investigated the growth inhibition effect of BEE on the HepG2 human hepatocellular carcinoma cell line. We performed a cell counting kit-8 assay to determine cell viability, and 4,6-diamino-2-phenylindole staining and flow cytometry to measure apoptotic cell death. Finally, the expression levels of pro- and anti-apoptotic proteins were measured. BEE inhibited the growth of HepG2 cells and induced apoptosis in a dose-dependent manner. Pro-apoptotic activity was promoted via the mitochondrial pathway of apoptosis, as demonstrated by the activation of pro-apoptotic proteins Bax, caspase-9, caspase-3, and poly (ADP-ribose) polymerase as well as the down-regulation of the anti-apoptotic protein Bcl-2. These results suggest that BEE may have potential use in hepatic cancer chemoprevention by suppressing cancer cell growth via pro-apoptotic activity.
Apoptosis/*drug effects ; Blotting, Western ; Carcinoma, Hepatocellular/*drug therapy/metabolism/pathology ; Caspase 3/metabolism ; Caspase 9/metabolism ; Cell Survival/drug effects ; Flow Cytometry ; Hep G2 Cells ; Humans ; Indoles/chemistry ; Liver Neoplasms/*drug therapy/metabolism/pathology ; Plant Bark/chemistry ; Plant Extracts/*pharmacology ; Poly(ADP-ribose) Polymerases/metabolism ; Ulmus/*chemistry ; bcl-2-Associated X Protein/metabolism

OBJECTIVETo evaluate the effect of Bear Bile Powder(, BBP) on the growth and apoptosis of HepG2 human hepatocellular carcinoma cells, and investigate the possible molecular mechanisms mediating its anti-cancer activity.

METHODSHepG2 cells were treated with 0.4-1.0 mg/mL of BBP for 24, 48 and 72 h. The viability of HePG2 cells was determined by MTT assay. Cellular morphology was observed via phase-contrast microscopy. Fluorescence-activated cell sorting analysis with Annexin-V/propidium idodide and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-benzimidazol-carbocyanine iodide (JC-1) staining was performed to determine cell apoptosis and the loss of mitochondrial membrane potential, respectively. Activation of caspase-9 and -3 was evaluated by a colorimetric assay.

RESULTSThe treatment with 0.4-1 mg/mL of BBP for 24, 48, or 72 h respectively reduced cell viability significantly by 7%-60%, 20%-90% or 25%-98%, compared with the untreated control cells (P<0.01). In addition, BBP treatment induced morphological changes in HepG2 cells. Furthermore, after treated with 0, 0.4, 0.6, 0.8 and 1.0 mg/mL of BBP, apoptosis cells (including early and late apoptotic cells) were 18.0%±1.3%, 34.9%±2.2%, 33.9%±2.8%, 37.4%±2.8% and 46.0%±2.5%, respectively (P<0.05); and the percentage of cells with reduced JC-1 red fluorescence were 6.6%±0.8%, 8.5%±0.8%, 13.5%±1.6%, 17.6%±2.3% and 46.7%±3.6%, respectively (P<0.01). Finally, BBP treatment significantly and dose-dependently induced activation of both caspase-9 and caspase-3 in HepG2 cells (P<0.05).

CONCLUSIONSBBP could inhibit the growth of HepG2 hepatocellular cancer cells through mitochondrion-mediated apoptosis, which may, in part, explain its anti-cancer activity. BBP may be a potential novel therapeutic agent for the treatment of hepatocellular carcinoma.

Animals ; Apoptosis ; drug effects ; Bile ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; Caspases ; metabolism ; Cell Proliferation ; drug effects ; Cell Shape ; drug effects ; Cell Survival ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hep G2 Cells ; Humans ; Liver Neoplasms ; drug therapy ; pathology ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondria ; drug effects ; metabolism ; Signal Transduction ; drug effects ; Ursidae

OBJECTIVE: To evaluate the effects of Celastrus Orbiculatus extracts (COE) on metastasis in hypoxia-induced hepatocellular carcinoma cells (HepG2) and to explore the underlying molecular mechanisms. METHODS: The effect of COE (160, 200 and 240 µ g/mL) on cell viability, scratch-wound, invasion and migration were studied by 3-4,5-dimethyl-2-thiazolyl-2,5-diphenyl-2-H-tetrazolium bromide (MTT), scratch-wound and transwell assays, respectively. CoCl was used to establish a hypoxia model in vitro. Effects of COE on the expressions of E-cadherin, vimentin and N-cadherin were investigated with Western blot and immunofluorescence analysis, respectively. RESULTS: COE inhibited proliferation and metastasis of hypoxia-induced hepatocellular carcinoma cells in a dose-dependent manner (P<0.01). Furthermore, the expression of epithelial-mesenchymal transition (EMT) related markers were also remarkably suppressed in a dose-dependent manner (P<0.01). In addition, the upstream signaling pathways, including the hypoxia-inducible factor 1 α (Hif-1 α) and Twist1 were suppressed by COE. Additionally, the Hif-1 α inhibitor 3-5'-hydroxymethyl-2'-furyl)-1-benzylindazole (YC-1), potently suppressed cell invasion and migration as well as expression of EMT in hypoxia-induced HepG2 cells. Similarly, the combined treatment with COE and YC-1 showed a synergistic effect (P<0.01) compared with the treatment with COE or YC-1 alone in hypoxia-induced HepG2 cells. CONCLUSIONS COE significantly inhibited the tumor metastasis and EMT by suppressing Hif-1 α/Twist1 signaling pathway in hypoxia-induced HepG2 cell. Thus, COE might have potential effect to inhibit the progression of HepG2 in the context of tumor hypoxia.
Biomarkers, Tumor ; metabolism ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; Celastrus ; chemistry ; Cell Hypoxia ; drug effects ; Cell Proliferation ; drug effects ; Cell Shape ; drug effects ; Cobalt ; Down-Regulation ; drug effects ; Epithelial-Mesenchymal Transition ; drug effects ; Hep G2 Cells ; Humans ; Liver Neoplasms ; drug therapy ; pathology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Proteins ; metabolism ; Plant Extracts ; pharmacology ; therapeutic use ; Signal Transduction ; drug effects