Blood Vessels ; chemistry ; physiopathology ; Carcinoma, Hepatocellular ; metabolism ; physiopathology ; Elasticity ; Humans ; Liver Neoplasms ; metabolism ; physiopathology ; Rupture, Spontaneous ; metabolism ; physiopathology ; von Willebrand Factor ; metabolism

BACKGROUND/AIMS: Tumor necrosis factor-alpha (TNF-alpha) exerts its anti-tumor effect through direct cytotoxicity on tumor cells and damage to the tumor vasculature. However, its role in tumor angiogenesis is controversial. We evaluated the angiogenic effect of TNF-alpha on BALB/c mouse colon carcinoma homograft model. METHODS: Ten BALB/c mice were inoculated intraperitoneally with CT-26 mouse colon carcinoma cells. After a week, recombinant mouse TNF-alpha (2microgram/mL) were given four times on every other day to five animals and the same volume of phosphate buffered saline was given at the same interval to five animals as control. Harvested tumor tissues were stained by immunohistochemistry with CD31 and VEGF antibodies. Number of microvessels and VEGF expression were counted by light microscope. RESULTS: The mean microvessel counts per 200x field of TNF-alpha treated animals were 70.2+/-7.8 and those of nontreated animals were 83.8+/-8.3 (p<0.05). The VEGF score of both groups were 3. CONCLUSIONS: TNF-alpha treated animals showed decreased microvessel counts in tumor tissue but VEGF expression in both groups showed no difference. Therefore, TNF-alpha showed antiangiogenic effects on colon carcinoma homograft model.
Animals ; Antigens, CD31/analysis ; Carcinoma/chemistry/*physiopathology ; Cell Line, Tumor ; Colonic Neoplasms/chemistry/*physiopathology ; English Abstract ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Neovascularization, Pathologic/*physiopathology ; Tumor Necrosis Factor-alpha/*pharmacology ; Vascular Endothelial Growth Factor A/analysis

To investigate the expression of microRNA (miRNA, miR) let-7e-3p in different cervical lesions and its clinical significance.The expression of miR-let-7e-3p in the tissues of normal cervix (=26), high-grade squamous intraepithelial lesion (HSIL) (=37), and cervix carcinoma (=101) were detected by reverse transcription and quantitative polymerase chain reaction (RT-qPCR). The correlation of miR-let-7e-3p expression with the clinicopathological parameters of patients with cervical cancer was analyzed. miR-let-7e-3p mimic was transfected into cervical carcinoma Siha cells. The cell cycle and apoptosis were determined by flow cytometry; cell proliferation was determined by CCK-8 kit; and the migration and invasion of cells were determined by Transwell assay.The relative expression levels of miR-let-7e-3p in normal cervix, HSIL, and cervical carcinoma were 1.45±0.24, 0.79±0.05 and 0.46±0.04, respectively (all<0.05). After transfection with miR-let-7e-3p mimic, the S-phase fraction and apoptosis rate of Siha cells were increased significantly compared with control group[(29.76±6.6)% vs (13.38±1.3)%,<0.05; (5.98±1.38)% vs (3.53±0.79)%,<0.05, respectively]. OD of transfected Siha cells at 48, 72 and 96 h were 0.57±0.11,0.65±0.04 and 0.84±0.14, which were significantly lower than those of untransfected Siha cells (0.74±0.05, 0.93±0.10 and 1.47±0.14, all<0.05). The migration and invasion abilities of transfected Siha cells were not significantly changed (all>0.05).The expression of miR-let-7e-3p is down-regulated in cervical neoplasms, which is associated with cell cycle arrest and proliferation inhibition of cervical cancer cells.
Apoptosis ; drug effects ; genetics ; Carcinoma ; chemistry ; genetics ; Cell Cycle ; drug effects ; genetics ; Cell Line, Tumor ; chemistry ; drug effects ; physiology ; Cell Movement ; drug effects ; genetics ; Cell Proliferation ; drug effects ; genetics ; Cervical Intraepithelial Neoplasia ; chemistry ; genetics ; physiopathology ; Down-Regulation ; physiology ; Female ; Humans ; MicroRNAs ; analysis ; pharmacology ; Neoplasm Invasiveness ; genetics ; physiopathology ; Neoplastic Processes ; Real-Time Polymerase Chain Reaction ; Transfection ; Uterine Cervical Neoplasms ; chemistry ; genetics ; physiopathology

OBJECTIVETo examine apoptosis of SMMC-7721 hepatocarcinoma cells induced by total flavonoids of Oxytropis falcata (TFOF) and its preliminary mechanism.

METHODSMMC-7721 cells were treated for 24 h with TFOF in different concentrations. Inhibition on proliferation of SMMC-7721 cells was assessed by MTT assay. The morphology of treated SMMC-7721 cells was observed by optical microscope. Effect of TFOF on the nuclear morphology of cells was analyzed using Hoechst 33258 staining by fluorescence microscope. Annexin V-FITC/PI staining and flow cytometric measurement were used for investigating the effect of TFOF on induction of apoptosis in SMMC-7721 cells and cell cycle analysis.

RESULTThe results of MTT assay showed that TFOF could induce cytotoxicity in SMMC-7721 cells in a dose-dependent manner. Hoechst 33258 staining analysis indicated that TFOF caused typical characteristics of apoptotic programmed cell death, such as cell shrinkage, apoptotic body formation etc. Flow cytometric analysis demonstrated that TFOF caused a dose-dependent apoptosis of SMMC-7721 cells and arrested cell cycle in G1 phase.

CONCLUSIONIt suggested that TFOF inhibit proliferation of SMMC-7721 cells by inducing apoptosis of the cells and arresting cell cycle in G1 phase.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; drug therapy ; physiopathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Liver Neoplasms ; drug therapy ; physiopathology ; Oxytropis ; chemistry

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

Despite septic arthritis is increasingly being reported in elderly patients with diabetes or alcoholism, reported cases of spontaneous bacterial arthritis in cirrhotic patients are extremely rare. We present the first reported case of K. pneumoniae septic arthritis and spontaneous bacterial peritonitis in a cirrhotic patient with hepatocellular carcinoma. K. pneumoniae, one of the most common causative organisms of spontaneous bacterial peritonitis in cirrhotic patients, was isolated from both the blood and the joint fluid, which suggests that the route of infection was hematogenous. After the treatment with cefotaxime and closed tube drainage, the condition of the patient was improved, and subsequently, the joint fluid became sterile and the blood cultures were proved negative. Therefore, this case provides further evidence for the mode of infection being bacteremia in cirrhotic patients and suggests that the enteric bacteremia in cirrhotics may cause infection in different organ systems.
Aged ; Animals ; Arthritis, Infectious/blood/*diagnosis/*microbiology ; *Carcinoma, Hepatocellular/pathology ; Fatal Outcome ; Female ; Humans ; Joints/chemistry/microbiology ; Klebsiella pneumoniae/*metabolism ; Liver Cirrhosis/*microbiology ; *Liver Neoplasms/pathology ; *Peritonitis/blood/microbiology/physiopathology

OBJECTIVETo evaluate the inhibitory effect of total bufadienolides from toad venom against H22 tumor in mice and preliminarily analyze the structures of the metabolites in tissues.

METHODHPLC and LC-MS were used for analysis of the chemical composition of TBFs. High, middle and low dosages of TBFs were orally administered or intra-peritoneally injected to H22 tumor-bearing mice for thirteen days. The animals were killed and the tumors were stripped and weighed. The metabolites in the tissues such as heart, liver, spleen, lung and kidney, were analyzed by HPLC and LC-MS.

RESULTThe chemical composition of TBFs were identified by comparison of the retention times with those of reference substances, on-line UV spectra and MS data. Its main components are concerned with gamabufotalin, arenobufagin, bufotalin, resibufagin, cinobufotalin, bufalin, cinobufagin and resibufogenin. TBFs had no obvious influence on body weight of H-22 tumor-bearing mice orally administered and the inhibition rate against tumor were 14.76%, 16.38% and 10.32% for low (5 mg x kg(-1)), middle (10 mg x kg(-1)) and high dosage (20 mg x kg(-1)), respectively. The mice intra-peritoneally injected with middle and high-dose of TBFs gained body weight slower than the control mice on the 5th day and recovered on the 13th day. The inhibition rate against tumor were 17.30%, 19.80% and 40.95% for low (1.5 mg x kg(-1)), middle (3 mg x kg(-1)) and high dose (6 mg x kg(-1)), respectively. The inhibitory effect took on dose-dependent manner. Based on the HPLC analyses on heart, liver, spleen, lung and kidney, bufadienolides were found in the liver tissue and 11 compounds of them were tentatively identified by LC-DAD-MS.

CONCLUSIONTBFs by oral administration had no inhibitory effect against H22 tumor in mice, however, TBFs by intra-peritoneal injection displayed the significantly inhibitory effect, accompanying some toxicity for early duration of the study. The identification of bufadienolides in the liver provides a good basis for the further investigation of the metabolic pathways of TBFs in vivo.

Amphibian Venoms ; chemistry ; Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; metabolism ; Bufanolides ; administration & dosage ; metabolism ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; physiopathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drug Administration Routes ; Female ; Humans ; Male ; Mice ; Mice, Inbred ICR ; Neoplasm Transplantation

OBJECTIVE: To investigate the expression of miR-33b in hepatocellular carcinoma (HCC) and to explore regulatory mechanism of miR-33b for cell proliferation of HCC.
 METHODS: HCC tissues and adjacent non-tumor tissues were collected for this study (n=32 for each). Real-time PCR and Western blot were conducted to examine the mRNA and protein expression, respectively. MTT assay was used to detect the cell proliferation. Luciferase reporter gene assay was performed to verify the target relationship between miR-33b and Sal-like 4 (SALL4).
 RESULTS: MiR-33b was significantly downregulated in HCC tissues compared with adjacent non-tumor tissues. Overexpression of miR-33b decreased the proliferation of HCC LH86 cells. SALL4 was identified as a target gene of miR-33b, and its protein expression was negatively regulated by miR-33b. Overexpression of SALL4 reversed the suppressive effect of miR-33b on LH86 cell proliferation. SALL4 was significantly upregulated in HCC tissues compared with adjacent non-tumor tissues.
 CONCLUSION The miR-33b suppresses HCC cell proliferation through down-regulation of SALL4.
Carcinoma, Hepatocellular ; chemistry ; genetics ; physiopathology ; Cell Proliferation ; genetics ; physiology ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; genetics ; physiology ; Humans ; Liver Neoplasms ; MicroRNAs ; analysis ; genetics ; physiology ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Transcription Factors ; genetics ; physiology ; Tumor Cells, Cultured ; Up-Regulation

To investigate the effect of silencing DJ-1 on xenografted human laryngeal squamous cell carcinoma (LSCC) Hep-2 cells in nude mice.Xenograft model of human LSCC was established by subcutaneous transplantation of Hep-2 cells in 24 nude mice. The LSCC-bearing nude mice were randomly divided into 3 groups (=8 in each):DJ-1 siRNA low dose group and DJ-1 siRNA high dose group were injected in tumors with 20 μg of DJ-1 siRNA or 40 μg of DJ-1 siRNA in 50 μL, respectively; control group was injected with 5% glucose solution in 50 μL, twice a week for 3 weeks. The weight and size of tumors were measured before injection. The animals were sacrificed 48 h after the final treatment, and the tumors were harvested and weighed. The apoptosis and proliferation of tumor cells were determined; the expressions of Caspase-3 and Ki-67 in tumor specimens were detected with immunohistochemistry. The expression of DJ-1, PTEN, survivin mRNA and protein in tumor tissues were detected by RT-PCR and Western blotting, respectively.Tumor weight in low dose group[(0.66±0.15)g] and high dose group[(0.48±0.11)g] were significantly lower than that in control group[(0.83±0.16)g, all<0.05]. The inhibition rates of low dose group and high dose group were (20.48±0.18)% and (42.16±0.13)%, respectively. Immunohistochemistry showed that the expression of Caspase-3 was increased and Ki-67 was reduced in tumor specimens, compared with the control group (all<0.05). RT-PCR and Western blot results showed that in low dose group and high dose group the mRNA and protein expression of DJ-1 and survivin significantly decreased (all<0.05), while PTEN mRNA and protein content increased (all<0.05).High dose DJ-1 siRNA can inhibit the tumor growth in human LSCC xenograft nude mouse model, which indicates that down-regulating DJ-1 and survivin, and up-regulating PTEN expression may lead to blockage of PI3K-PKB/Akt signaling pathway and promoting tumor cell apoptosis.
Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; genetics ; Carcinoma, Squamous Cell ; chemistry ; genetics ; physiopathology ; Caspase 3 ; analysis ; drug effects ; Cell Line, Tumor ; chemistry ; drug effects ; physiology ; transplantation ; Cell Proliferation ; drug effects ; Down-Regulation ; Gene Expression Regulation ; drug effects ; genetics ; physiology ; Head and Neck Neoplasms ; chemistry ; genetics ; physiopathology ; Heterografts ; drug effects ; physiology ; Humans ; Inhibitor of Apoptosis Proteins ; analysis ; drug effects ; Ki-67 Antigen ; analysis ; drug effects ; Laryngeal Neoplasms ; chemistry ; genetics ; physiopathology ; Mice, Nude ; PTEN Phosphohydrolase ; analysis ; drug effects ; Phosphatidylinositol 3-Kinases ; drug effects ; Protein Deglycase DJ-1 ; pharmacology ; Proto-Oncogene Proteins c-akt ; drug effects ; RNA Interference ; physiology ; RNA, Messenger ; pharmacology ; RNA, Small Interfering ; physiology ; Signal Transduction ; drug effects ; genetics ; physiology

OBJECTIVETo observe the effect of anti-tumor and mechanism of the extract of Spatholobus suberctus (SSCE) in vivo.

METHODThe mouse model of Lewis lung carcinoma was used to investigate the effects of SSCE on tumor growth and metastasis. Furthermore, we explored the mechanism of anti-tumor by analyze the cell cycle and determine the apoptosis.

RESULTThe studies demonstrated that the tumor inhibitory rate of SSCE in low dose group was the highest (30.65%) on Lewis lung cancer. SSCE can resist metastasis, at the same time, it can induce cell cycle arrested in G1 phase, whereas, there was no significant difference in apoptotic rate each group.

CONCLUSIONWe verified that SSCE exits anti-tumor effect and resist metastasis, furthermore, it can arrest function cell in G1 phase.

Animals ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Carcinoma, Lewis Lung ; drug therapy ; physiopathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Disease Models, Animal ; Fabaceae ; chemistry ; Humans ; Lung Neoplasms ; drug therapy ; physiopathology ; Male ; Mice ; Mice, Inbred C57BL ; Plant Extracts ; pharmacology ; therapeutic use