BACKGROUNDHepatocellular carcinoma (HCC) is one of the deadliest cancers worldwide. In order to investigate the molecular biologic mechanism of HCC's development, we studied the expressions of SE-1, CD105 and CD31 in tumor endothelial cells (TECs) of HCC and in the serum of rats.

METHODSWe analyzed the expressions of SE-1, CD31 and CD105 in rat HCC tumor tissues using immunohistochemistry (IHC). Twenty HCC bearing rats and eighteen normal rats were examined for the expressions of SE-1, CD31 and CD105 antigens in serum by enzyme-linked immunosorbent assay (ELISA).

RESULTSSE-1, CD31 and CD105 antigens were detected both in HCC tissue and in normal liver tissue with higher expressions of CD31 and CD105 in HCC while the SE-1 antigen expression was higher in normal liver. Similarly, serum CD31 and CD105 in rats with HCC were significantly increased compared with normal rats (t = 2.8628, P = 0.0086; t = 4.4922, P < 0.0001, respectively). In contrast, SE-1 antigen in HCC rat serum was significantly decreased compared with normal rats (t = 3.4983, P = 0.0011).

CONCLUSIONSE-1, CD31 and CD105 are closely related with liver tumor angiogenesis, which is similar to their performances in terms of their expressions in the serum.

Animals ; Antigens, CD ; blood ; Carcinoma, Hepatocellular ; blood supply ; chemistry ; Endothelial Cells ; chemistry ; immunology ; Enzyme-Linked Immunosorbent Assay ; Immunohistochemistry ; Liver Neoplasms, Experimental ; blood supply ; chemistry ; Male ; Neovascularization, Pathologic ; blood ; Platelet Endothelial Cell Adhesion Molecule-1 ; blood ; Rats ; Rats, Inbred BUF

BACKGROUND/AIMS: Tumor angiogenesis, a major requirement for tumor growth and metastasis, is regulated by pro- and anti-angiogenic factors. Hepatocellular carcinoma (HCC) has become a common malignant tumor worldwide. It is characterized by a high vascularity. METHODS: We studied the immunohistochemical expression of angiostatin, vascular endothelial cell growth factor (VEGF), matrix metalloproteinase (MMP)-9 and MMP-12, and the relationship between these results and the microvessel density (MVD) in 48 HCC specimens. To determine whether HCC cells express angiostatin per se, we examined the expression of angiostatin, MMP-9 and MMP-12 by Western blotting in four HCC cell lines. RESULTS: Expression of angiostatin and MMP-12 (but not MMP-9) were strongly correlated with decreased MVD in HCCs (P=0.006, P=0.038, respectively). VEGF positive tumors showed a significantly higher MVD than VEGF negative tumors (P=0.01). We divided the 48 cases into the following four groups: group A, angiostatin (+), MMP-9 or -12 (+), and VEGF (-); group B, angiostatin (-) and VEGF (-); group C, angiostatin (+), MMP-9 or -12 (+), and VEGF (+); group D, angiostatin (-) and VEGF (+). There was a significant correlation with MVD among these groups (P<0.001). Angiostatin was detected by Western blotting in 2 out of 4 HCC cell lines and was associated with plasminogen and MMP expression. CONCLUSIONS: These results indicate that angiogenesis in HCC is a complex process involving multiple factors including angiostatin, VEGF, and MMP. Our results suggest that angiostatin is generated by MMP-mediated proteolysis of plasminogen in HCC cells.
Adult ; Aged ; Angiogenesis Inhibitors/analysis/physiology ; Angiostatins/analysis/physiology ; Carcinoma, Hepatocellular/*blood supply/chemistry ; English Abstract ; Female ; Gelatinase B/analysis/physiology ; Humans ; Liver Neoplasms/*blood supply/chemistry ; Male ; Metalloendopeptidases/analysis/physiology ; Middle Aged ; Neovascularization, Pathologic/metabolism/*physiopathology ; Vascular Endothelial Growth Factor A/analysis/physiology

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

OBJECTIVETo detect the biological factor association with metastasis and recurrence of hepato cellular carcinoma (HCC).

METHODSLabeled streptavidin biotin method was performed to study VEGF, uPA and ICAM-1 protein, and antigen of PCNA expression in 123 patients with HCC. Venous invasion was observed under microscope at the same time.

RESULTSThe expression rate of VEGF was higher in HCC with intra-hepatic metastasis in group B than in HCC without PVTT/metastasis in group A (P < 0.01) and higher in HCC with PVTT in group C and PVTT in group D higher than in group B (P < 0.05). The expression rate of uPA protein was higher in group B than in group A (P < 0.01), but no significant difference in groups B and C. The expression rate of ICAM-1 showed no significant difference in the four groups. MVD and PCNA-LI increased gradually from group A to D. The rate of microscopic venous invasion in group B was higher than in group A (P < 0.05), in group D higher than in group B (P < 0.05), but no significant difference was observed between groups B and C, groups C and D (P = 0.16, 0.13 respectively). The rate of postoperative recurrence of HCC was higher in group B than in group A, and lower than in group C. Multivariate regression analysis: showed postoperative recurrence was correlated very well with microscopic venous invasion (r = 0.783, P < 0.01), and MVD (r = 0.143, P < 0.05). Metastasis of HCC were associated very well with PCNA-LI (r = 0.590, P < 0.01) and MVD (r = 0.179, P < 0.05), and negatively correlated with the rate of ICAM-1 expression (r = -0.183, P < 0.01).

CONCLUSIONSVEGF, uPA and ICAM-1 protein expression and proliferation of cancer cells could contribute to the formation of PVTT, metastasis and postoperative recurrence of HCC. Over-proliferated cancer cells in HCC could be the direct factor of intrahepatic metastasis and formation of PVTT, and microscopic venous invasion may be a significant factor to predict postoperative recurrence of HCC.

Carcinoma, Hepatocellular ; blood supply ; pathology ; secondary ; Endothelial Growth Factors ; analysis ; Humans ; Immunohistochemistry ; Intercellular Adhesion Molecule-1 ; analysis ; Intercellular Signaling Peptides and Proteins ; analysis ; Liver Neoplasms ; blood supply ; chemistry ; pathology ; Lymphokines ; analysis ; Neoplasm Recurrence, Local ; Proliferating Cell Nuclear Antigen ; analysis ; Urokinase-Type Plasminogen Activator ; analysis ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

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