No abstract available.
Carcinoma, Hepatocellular/*blood supply/metabolism/*therapy ; Cell Line ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit/*antagonists & ; Liver Neoplasms/*blood supply/metabolism/therapy ; Neovascularization, Pathologic/genetics/metabolism/*therapy ; RNA Interference ; RNA, Small Interfering/metabolism

BACKGROUND/AIMS: Hypoxia-inducible factor-1alpha (HIF-1alpha) is a central transcriptional factor involved in the cellular responses related to various aspects of cancer biology, including proliferation, survival, and angiogenesis, and the metabolism of the extracellular matrix in hypoxia. This study evaluated whether adenovirus-mediated small hairpin RNA (shRNA) against HIF-1alpha (shHIF-1alpha) inhibits cell proliferation and angiogenesis in hepatocellular carcinoma (HCC) cell lines. METHODS: Knockdown of HIF-1alpha expression was constructed by adenovirus-mediated RNA interference tools, and HCC cell lines infected with shHIF-1alpha coding virus were cultured under a hypoxia condition (1% O2) for 24 hours. Following infection, the expression levels of HIF-1alpha, angiogenesis factors, and matrix metalloproteinase (MMP) were examined using Western blotting. Cell proliferation and angiogenesis were measured by a cell proliferation assay (MTT assay) and an angiogenesis-related assay (invasion and tube-formation assay), respectively. RESULTS: Adenovirus mediated inhibition of HIF-1alpha induced suppression of tumor growth in HCC cell lines. It also down-regulated the expression of angiogenesis factor and MMP proteins. Angiogenesis as well as mobility of vascular cells to tumor was suppressed by adenovirus-mediated shHIF-1alpha-infected groups in human umbilical vein endothelial cells (HUVECs). CONCLUSIONS: These data suggest that adenovirus-mediated inhibition of HIF-1alpha inhibits the invasion, tube formation, and cell growth in HUVECs and HCC cells.
Adenoviridae/genetics ; Carcinoma, Hepatocellular/*blood supply/metabolism/therapy ; Cell Line, Tumor ; Cell Proliferation ; Endothelial Cells/metabolism ; Gene Knockdown Techniques ; Genetic Vectors ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit/*antagonists & ; Liver Neoplasms/*blood supply/metabolism/therapy ; Matrix Metalloproteinases/metabolism ; Neovascularization, Pathologic/genetics/metabolism/*therapy ; RNA Interference ; RNA, Small Interfering/metabolism

OBJECTIVETo observe the clinical combination effect of Jinlong Capsule (JLC) and transcatheter arterial chemoembolization (TACE) on the patients with primary hepatic carcinoma (PHC) and JLC' s influence on serum osteopontin (OPN) expression and elucidate the correlation between the serum OPN level and curative effect of JLC and TACE.

METHODSA total of 98 patients with PHC were observed in a randomized controlled trial (RCT). They were assigned to the Chinese medicine (CM) group (53 patients who were treated with TACE and JLC) and the intervention group (45 patients who were treated with TACE only). The serum OPN levels were measured before and after treatment by quantitative sandwich enzyme-linked immunosorbent assay (ELISA). Forty healthy people were assigned to the control group. The clinical efficacy was observed and Karnofsky score (KPS) was graded.

RESULTSThe clinical efficacy of the CM group (60.38%) was better than that of the intervention group (40.00 %), and the KPS (84.35+/-12.19) was higher than the intervention group (69.86+/- 11.58) (P<0.05). The serum OPN levels before and after treatment in the patients with PHC were significantly elevated compared with those in the control group (P<0.01). After treatment, the OPN levels in CM group (117.69 <+/-78.50) were significantly lower compared with those in intervention group (151.09+/-83.90, P<0.05). The OPN levels of responders were remarkably lowered than the non-responders after treatment, and the level of OPN in the CM group was lower than the intervention group (P<0.05).

CONCLUSIONSThe short-term clinical efficacy and the quality of life of patients with PHC can be improved by combining JLC with TACE. The serum OPN levels in PHC patients can reflect the curative effect of treatment and the prognosis of the disease.

Adult ; Aged ; Antineoplastic Agents, Phytogenic ; administration & dosage ; Capsules ; Carcinoma, Hepatocellular ; blood ; metabolism ; therapy ; Catheterization, Peripheral ; methods ; Chemoembolization, Therapeutic ; methods ; Combined Modality Therapy ; Drugs, Chinese Herbal ; administration & dosage ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Liver ; blood supply ; Liver Neoplasms ; blood ; metabolism ; therapy ; Male ; Middle Aged ; Osteopontin ; analysis ; blood ; metabolism ; Time Factors ; Treatment Outcome

Angiogenesis Inhibitors ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Hepatocellular ; blood supply ; metabolism ; pathology ; therapy ; Chemoembolization, Therapeutic ; methods ; Endostatins ; therapeutic use ; Humans ; Liver Neoplasms ; blood supply ; metabolism ; pathology ; therapy ; Microvessels ; pathology ; Neovascularization, Pathologic ; pathology ; Vascular Endothelial Growth Factors ; metabolism

OBJECTIVETo evaluate the antiangiogenic property of pigment epithelium-derived factor(PEDF) in heptocarcinoma cell lines and explore its possible application in the gene therapy of human hepatocellular carcinoma (HCC).

METHODSThe gene encoding human PEDF was subcloned into lentiviral vector to generate the recombinant plasmid pLenti-PEDF. The plasmid pLenti-PEDF and two other packaging plasmids were cotransfected to 293T cells by calcium phosphate. Then HepG2 was infected with recombinant lentivirus and the expression efficiency of PEDF was analyzed by western blot. Proliferation and migration assay of human umbilical vein endothelial cells (HUVEC) was used to evaluate the biological activity of PEDF in vitro. Murine subcutaneous tumor model was established to investigate the therapeutic effects of Lenti-PEDF on HCC, and the expression of PEDF mRNA in tumor tissues was analyzed by RT-PCR.

RESULTSRestriction enzyme digestion and DNA sequencing demonstrated that the recombinant plasmid pLenti-PEDF was constructed successfully. HepG2 secreted PEDF in the media effectively after infected with the recombinant lentivirus and this protein exhibited strong inhibitory effects on proliferation and migration of human umbilical vein endothelial cells (P less than 0.01). Intratumoral injection of Lenti-PEDF caused significant inhibition of tumor growth (P less than 0.01), and high level expression of PEDF mRNA was detected in tumor tissues by RT-PCR.

CONCLUSIONSOur data suggest that PEDF may exert an inhibitory effect on tumor angiogenesis and PEDF gene therapy may provide a new approach for the treatment of HCC.

Animals ; Cell Proliferation ; Endothelial Cells ; metabolism ; Eye Proteins ; genetics ; metabolism ; Genetic Therapy ; Genetic Vectors ; Hep G2 Cells ; Humans ; Lentivirus ; genetics ; Liver Neoplasms ; blood supply ; genetics ; therapy ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neovascularization, Pathologic ; therapy ; Nerve Growth Factors ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Serpins ; genetics ; metabolism ; Transfection ; Umbilical Veins ; cytology

OBJECTIVEThe efficiency network is a complicated network for revealing the efficient mechanism of traditional Chinese medicines (TCMs) and relations among efficiencies. The efficiency-property relations were used to establish a warm-pungent-liver efficiency network to explain the principle of treating hepatoma with medicines invigorating blood circulation and eliminating blood stasis. Safflower, a warm-pungent medicine distributing along the live meridian, was taken for example to discuss the efficiency network' s application in the identification of active ingredients of TCMs and the combination.

METHODIn the early stage of this study, combined warm-pungent-liver medicines distributed along the liver meridian and invigorating blood circulation and eliminating blood stasis were taken as the study objects to collect the pharmacological effect data of warm-pungent-liver medicines and obtain the pharmacological effect combinations with the highest blood circulation-invigorating association by the association rules and the chi-square test. The pharmacological target data recorded in the DrugBank database is used to establish the warm-pungent-liver efficiency network according to the principle line of "efficiency-property-pharmacology-target-protein interaction" under the background of the protein interaction network.

RESULTThe blood circulation-invigorating medicines could directly treat hepatoma by impacting protooncogene, cancer suppressor gene, cell apoptosis and anti-inflammation, and indirectly treat hepatoma by resisting coagulation and adhesion, regulating local blood circulation, preventing cancer cell metastasis and enhancing the tissues' sensitivity to the anticancer drugs. Among the active ingredients of safflower screened based on the blood circulation-invigorating network targets, carthamin yellow, quercetin and luteolin have been proved to have the anti-hepatoma effect in literatures, which indicated the reliability of this study's results and the purpose of the efficiency network.

CONCLUSIONThe efficiency network is an effective method for revealing the TCM's mechanism, and lays a foundation for discovering key active ingredients of TCMs for treating specific diseases.

Antineoplastic Agents, Phytogenic ; chemistry ; therapeutic use ; Blood Circulation ; drug effects ; Carcinoma, Hepatocellular ; drug therapy ; genetics ; metabolism ; physiopathology ; Drugs, Chinese Herbal ; chemistry ; therapeutic use ; Gene Regulatory Networks ; drug effects ; Humans ; Liver ; blood supply ; drug effects ; Liver Neoplasms ; drug therapy ; genetics ; metabolism ; physiopathology

Hepatocellular carcinoma (HCC) is a major global health problem, which has a grave morbidity and mortality. Over the past few decades, no effective systemic therapeutic modalities have been established for patients with the unresectable HCC in advanced stage. Sorafenib is a small molecule that blocks cancer cell proliferation by targeting the intracellular signaling pathway at the level of Raf-1 and B-Raf serine-threonine kinases, and exerts an anti-angiogenic effect by targeting the vascular endothelial growth factor receptor-1, 2 and 3, and platelet-derived growth factor receptor-beta tyrosine kinases. Recently, two clinical successful applications, SHARP and Asia-Pacific trial, of multikinase inhibitor sorafenib represent a significant advance in the treatment of advanced HCC patients without a curative chance. However, because the results of clinical trials show diverse responses in a subset of HCC patients, a molecular classification of HCC through the excavation of specific biomarkers related to its biological behavior is necessary for sorting HCC patients to each group with a biological homogeneity, ultimately leading to the most suitable individualization of molecular targeted therapy in HCC.
Antineoplastic Agents/therapeutic use ; Benzenesulfonates/therapeutic use ; Carcinoma, Hepatocellular/pathology/secondary/*therapy ; Humans ; Liver Neoplasms/blood supply/pathology/*therapy ; Neovascularization, Pathologic ; Proto-Oncogene Proteins B-raf/antagonists & inhibitors/metabolism ; Proto-Oncogene Proteins c-raf/antagonists & inhibitors/metabolism ; Pyridines/therapeutic use ; Receptors, Platelet-Derived Growth Factor/antagonists & inhibitors/metabolism ; Receptors, Vascular Endothelial Growth Factor/antagonists & inhibitors/metabolism ; Signal Transduction

OBJECTIVETo investigate the role of metronomic chemotherapy of S-1 on angiogenesis of hepatocellular carcinoma in animal model.

METHODS-1 was dissolved in a 0.5% (w/v) HPMC solution. 30 LCI-D20 were randomly devided into five groups: control group(O), 10 mg * kg(-1) * d(-1) S-1 group (A), 1 mg * kg(-1) * d(-1) S-1 group (B), 0.5 mg * kg(-1) * d(-1) S-1 group (C) and 0.25 mg * kg(-1) * d S-1 group (D). 28 days after the treatment with 0.5% (w/v) HPMC solution, tumors in LCI-D20 mice were moved out. Tumor mass was measured and microvessel density (MVD) was used to evaluate angiogenesis in tumor. The cellular apoptosis was determined using flow cytometry. The expression of VEGF, bFGF and TSP-1 was measured by RT-PCR.

RESULTSThe mean tumor mass was 2.01, 0.38, 1.12, 1.38, 2.27 g in O, A, B, C, D group, respectively. The mean MVD was 39.57, 19.90, 5.93, 17.10, 29.53 in O, A, B, C, D respectively. The mean tumor cellular apoptosis rate was 4.08%, 44.37%, 31.73%, 19.83%, and 8.25% in O, A, B, C, D respectively. The expression of VEGF and bFGF in O group was highest, and A was slightly low, and C and D taked the third place, and B was the lowst; The expression of TSP-1 in B was highest, and C and D were slightly low, and A taked the third place, and O was the lowst.

CONCLUSIONMetronomic chemotherapy of S-1 destabilizes pre-existing tumor vasculature and inhibits ongoing angiogenesis.

Animals ; Antimetabolites, Antineoplastic ; therapeutic use ; Apoptosis ; Carcinoma, Hepatocellular ; blood supply ; drug therapy ; pathology ; Drug Combinations ; Fibroblast Growth Factor 2 ; genetics ; metabolism ; Liver Neoplasms, Experimental ; blood supply ; drug therapy ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neovascularization, Pathologic ; Oxonic Acid ; administration & dosage ; therapeutic use ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Reverse Transcriptase Polymerase Chain Reaction ; Tegafur ; administration & dosage ; therapeutic use ; Vascular Endothelial Growth Factors ; genetics ; metabolism

OBJECTIVETo study the effects of sirolimus (SRL) on the growth of transplanted human hepatocellular carcinoma (HCC) in nude mice.

METHODSHepG2 cells were Implanted into the liver of nude mice. The implanted mice were then treated with SRL and tacrolimus (FK506). The expression of vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) was detected by immunohistology, microvessel density (MVD) was counted by immunostaining with anti-CD34 antibody for endothelial cells. Tumor apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay.

RESULTSThe tumor weight was (352+/-38) mg, (683+/-53) mg and (675+/-45) mg in SRL, FK506 and control group respectively. The tumor weight was significantly decreased in SRL group (P < 0.01), and there was no difference between FK506 group and control group. The expression of VEGF and PCNA protein was remarkably down-regulated in SRL group compared to control group (P < 0.05), and it was not significantly different between FK506 group and control group (P > 0.05). Compared to the control group, MVD was significanly decreased in SRL group, and the apoptosis index of tumor cell was significantly higher in SRL group (P < 0.01).

CONCLUSIONSRL inhibits transplanted HCC tumor growth by reducing tumor angiogenesis, inhibiting tumor proliferation and inducing tumor apoptosis.

Animals ; Antineoplastic Agents ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; pathology ; Hep G2 Cells ; Humans ; Immunohistochemistry ; Liver ; blood supply ; pathology ; Liver Neoplasms, Experimental ; drug therapy ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neovascularization, Pathologic ; prevention & control ; Proliferating Cell Nuclear Antigen ; metabolism ; Sirolimus ; administration & dosage ; pharmacology ; Tacrolimus ; administration & dosage ; pharmacology ; Treatment Outcome ; Vascular Endothelial Growth Factor A ; metabolism ; Xenograft Model Antitumor Assays

OBJECTIVECelastrus orbiculatus Thunb. has been used for thousands of years in China as a remedy against cancer and inflammatory diseases. This study aims to investigate whether C. orbiculatus extract (COE) could inhibit angiogenesis, which is the pivotal step in tumor growth, invasiveness, and metastasis.

METHODSIn this study, the extract from the stem of C. orbiculatus was used. Mouse hepatic carcinoma cells (Hepa1-6) were treated with COE in different nontoxic concentrations (10, 20, 40, 80, and 160 μg/mL). The mRNA and protein expression levels of vascular endothelial growth factor (VEGF) were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot, respectively; the active fractions were further tested on C57BL/6 mice and human umbilical vein endothelial cells (HUVEC) for any antiangiogenic effects.

RESULTSCOE significantly inhibited proliferation and induced apoptosis in Hepa1-6 cells and inhibited VEGF expression at both mRNA and protein levels. Furthermore, this agent inhibited the formation of the capillary-like structure in primary cultured HUVEC in a dose-dependent manner. In vivo, COE significantly reduced the volume and weight of solid tumors with low adverse effects and decreased tumor angiogenesis.

CONCLUSIONSIn summary, COE could be used to treat hepatic carcinoma. The mechanisms of the antitumor activity of COE may be due to its effects against tumor angiogenesis by targeting the VEGF protein.

Administration, Oral ; Angiogenesis Inhibitors ; pharmacology ; therapeutic use ; Animals ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; blood supply ; drug therapy ; pathology ; Celastrus ; chemistry ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Collagen ; metabolism ; Drug Combinations ; Human Umbilical Vein Endothelial Cells ; drug effects ; Humans ; Laminin ; metabolism ; Liver Neoplasms ; blood supply ; drug therapy ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Neovascularization, Pathologic ; drug therapy ; pathology ; Neovascularization, Physiologic ; drug effects ; Phytotherapy ; Plant Extracts ; administration & dosage ; pharmacology ; therapeutic use ; Plant Stems ; chemistry ; Proteoglycans ; metabolism ; Signal Transduction ; drug effects ; Transcriptional Activation ; drug effects ; genetics ; Tumor Burden ; drug effects ; Vascular Endothelial Growth Factor A ; biosynthesis ; metabolism