BACKGROUND/OBJECTIVES: Abundant consumption of seaweeds in the diet is epidemiologically linked to the reduction in risk of developing cancer. In larger cases, however, identification of particular seaweeds that are accountable for these effects is still lacking, hindering the recognition of competent dietary-based chemo preventive approaches. The aim of this research was to establish the antiproliferative potency and angiosuppressive mode of action of Stoechospermum marginatum seaweed methanolic extract using various experimental models. MATERIALS/METHODS: Among the 15 seaweeds screened for antiproliferative activity against Ehrlich ascites tumor (EAT) cell line, Stoechospermum marginatum extract (SME) was found to be the most promising. Therefore, it was further investigated for its anti-proliferative activity in-vitro against choriocarcinoma (BeWo) and non-transformed Human embryonic kidney (HEK 293) cells, and for its anti-migratory/tube formation activity against HUVEC cells in-vitro. Subsequently, the angiosuppressive activity of S. marginatum was established by inhibition of angiogenesis in in-vivo (peritoneal angiogenesis and chorioallantoic membrane assay) and ex-vivo (rat cornea assay) models. RESULTS: Most brown seaweed extracts inhibited the proliferation of EAT cells, while green and red seaweed extracts were much less effective. According to the results, SME selectively inhibited proliferation of BeWo cells in-vitro in a dose-dependent manner, but had a lesser effect on HEK 293 cells. SME also suppressed the migration and tube formation of HUVEC cells in-vitro. In addition, SME was able to suppress VEGF-induced angiogenesis in the chorio allantoic membrane, rat cornea, and tumor induced angiogenesis in the peritoneum of EAT bearing mice. A decrease in the microvessel density count and CD31 antigen staining of treated mice peritoneum provided further evidence of its angiosuppressive activity. CONCLUSIONS: Altogether, the data underline that VEGF mediated angiogenesis is the target for the angiosuppressive action of SME and could potentially be useful in cancer prevention or treatment involving stimulated angiogenesis.
Allantois ; Animals ; Antigens, CD31 ; Carcinoma, Ehrlich Tumor ; Cell Line ; Chorioallantoic Membrane ; Choriocarcinoma ; Cornea ; Diet ; Female ; HEK293 Cells ; Human Umbilical Vein Endothelial Cells ; Humans ; Kidney ; Methanol ; Mice ; Microvessels ; Models, Theoretical* ; Peritoneum ; Pregnancy ; Rats ; Seaweed ; Vascular Endothelial Growth Factor A

BACKGROUND/OBJECTIVES: Abundant consumption of seaweeds in the diet is epidemiologically linked to the reduction in risk of developing cancer. In larger cases, however, identification of particular seaweeds that are accountable for these effects is still lacking, hindering the recognition of competent dietary-based chemo preventive approaches. The aim of this research was to establish the antiproliferative potency and angiosuppressive mode of action of Stoechospermum marginatum seaweed methanolic extract using various experimental models. MATERIALS/METHODS: Among the 15 seaweeds screened for antiproliferative activity against Ehrlich ascites tumor (EAT) cell line, Stoechospermum marginatum extract (SME) was found to be the most promising. Therefore, it was further investigated for its anti-proliferative activity in-vitro against choriocarcinoma (BeWo) and non-transformed Human embryonic kidney (HEK 293) cells, and for its anti-migratory/tube formation activity against HUVEC cells in-vitro. Subsequently, the angiosuppressive activity of S. marginatum was established by inhibition of angiogenesis in in-vivo (peritoneal angiogenesis and chorioallantoic membrane assay) and ex-vivo (rat cornea assay) models. RESULTS: Most brown seaweed extracts inhibited the proliferation of EAT cells, while green and red seaweed extracts were much less effective. According to the results, SME selectively inhibited proliferation of BeWo cells in-vitro in a dose-dependent manner, but had a lesser effect on HEK 293 cells. SME also suppressed the migration and tube formation of HUVEC cells in-vitro. In addition, SME was able to suppress VEGF-induced angiogenesis in the chorio allantoic membrane, rat cornea, and tumor induced angiogenesis in the peritoneum of EAT bearing mice. A decrease in the microvessel density count and CD31 antigen staining of treated mice peritoneum provided further evidence of its angiosuppressive activity. CONCLUSIONS: Altogether, the data underline that VEGF mediated angiogenesis is the target for the angiosuppressive action of SME and could potentially be useful in cancer prevention or treatment involving stimulated angiogenesis.
Allantois ; Animals ; Antigens, CD31 ; Carcinoma, Ehrlich Tumor ; Cell Line ; Chorioallantoic Membrane ; Choriocarcinoma ; Cornea ; Diet ; Female ; HEK293 Cells ; Human Umbilical Vein Endothelial Cells ; Humans ; Kidney ; Methanol ; Mice ; Microvessels ; Models, Theoretical* ; Peritoneum ; Pregnancy ; Rats ; Seaweed ; Vascular Endothelial Growth Factor A

OBJECTIVETo determine the hepatoprotective effect of acetone semicarbazone (ASC) in vivo in normal and Ehrlich ascites carcinoma (EAC) bearing male Swiss albino mice.

METHODSDrug-induced changes in biochemical and behavioral parameters at dose of 2.0 mg/kg body weight for 14 d and nullifying the toxicity induced by EAC cells were studied. The histopathology studies of the protective effects of ASC on vital organs were also assessed.

RESULTSThe administration of ASC made insignificant changes in body weight and behavioral (salivation, diarrhea, muscular numbness) changes during treatment period due to minor toxicity were minimized after the treatment in normal mice. The biochemical parameters, including serum glutamate pyruvate transaminase, glutamate oxaloactate transaminase, alkaline phosphatase, serum glucose, cholesterol, urea, triglyceride and billirubin changed modestly in normal mice receiving ASC. Though the treatment continued, these values gradually decreased to normal level after the treatment. In EAC bearing mice, the toxic effects due to EAC cells in all cases were nullified by treatment with the ASC. Significant abnormalities were not detected in histology of the various organs of the normal mice treated with ASC.

CONCLUSIONSASC can, therefore, be considered safe in formulating novel anticancer drug, as it exhibits strong protective effect against EAC cell bearing mice.

Acetone ; analogs & derivatives ; pharmacology ; therapeutic use ; Animals ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Carcinogenesis ; drug effects ; Carcinoma, Ehrlich Tumor ; drug therapy ; Liver ; drug effects ; Male ; Mice ; Semicarbazones ; pharmacology ; therapeutic use

Cancer cell vaccine-based immunotherapy has received increasing interest in many clinical trials involving patients with breast cancer. Combining with appropriate adjuvants can enhance the weak immunogenic properties of tumor cell lysates (TCL). In this study, diphtheria toxin (DT) and two tandem repeats of mycobacterial heat shock protein 70 (mHSP70) fragment 407-426 (M2) were conjugated to TCL with glutaraldehyde, and the constructed cancer cell vaccine was named DT-TCL-M2. Subcutaneous injection of DT-TCL-M2 in mice effectively elicited tumor-specific polyclonal immune responses, including humoral and cellular immune responses. High levels of antibodies against TCL were detected in the serum of immunized mice with ELISA and verified with Western blot analyses. The splenocytes from immunized mice showed potent cytotoxicity on Ehrlich ascites carcinoma cells. Moreover, the protective antitumor immunity induced by DT-TCL-M2 inhibited tumor growth in a mouse breast tumor model. DT-TCL-M2 also attenuated tumor-induced angiogenesis and slowed tumor growth in a mouse intradermal tumor model. These findings demonstrate that TCL conjugated with appropriate adjuvants induced effective antitumor immunity in vivo. Improvements in potency could further make cancer cell vaccines a useful and safe method for preventing cancer recurrence after resection.
Adjuvants, Immunologic ; Animals ; Bacterial Proteins ; genetics ; immunology ; Cancer Vaccines ; immunology ; Carcinoma, Ehrlich Tumor ; immunology ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Diphtheria Toxin ; genetics ; immunology ; Female ; HSP70 Heat-Shock Proteins ; genetics ; immunology ; Immunoglobulin G ; immunology ; Immunotherapy ; Mice ; Neovascularization, Pathologic ; Peptide Fragments ; genetics ; immunology ; Recombinant Fusion Proteins ; genetics ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Tandem Repeat Sequences

OBJECTIVETo evaluate the antitumor activity of Manilkara zapota (M. zapota) L. stem bark against Ehrlich ascites carcinoma (EAC) in Swiss albino mice.

METHODSThe in vivo antitumour activity of the ethyl acetate extract of stem bark of M. zapota L. (EASM) was evaluated at 50, 100 and 200 mg/kg bw against EAC using mean survival time. After administration of the extract of M. zapota, viable EAC cell count and body weight in the EAC tumour hosts were observed. The animal was also observed for improvement in the haematological parameters (e.g., heamoglobin content, red and white blood cells count and differential cell count) after EASM treatment.

RESULTSIntraperitoneal administration of EASM reduced viable EAC cells, increased the survival time, and restored altered haematological parameters. Significant efficacy was observed for EASM at 100 mg/kg dose (P<0.05).

CONCLUSIONSIt can be concluded that the ethyl acetate extract of stem bark of M. zapota L. possesses significant antitumour activity.

Animals ; Antineoplastic Agents ; isolation & purification ; therapeutic use ; Body Weight ; Carcinoma, Ehrlich Tumor ; drug therapy ; Cell Survival ; drug effects ; Disease Models, Animal ; Injections, Intraperitoneal ; Male ; Manilkara ; chemistry ; Mice ; Plant Bark ; chemistry ; Plant Extracts ; isolation & purification ; therapeutic use ; Survival Analysis ; Treatment Outcome

OBJECTIVETo investigate the effect of endostar in controlling ascites tumor formation in mice.

METHODSMouse models bearing ascites tumors were established via intraperitoneal injection of H22 and S180 cell lines. Eighty-eight ICR mice were randomly assigned into 4 groups, namely the control group (0.9% normal saline) and 3 endostar groups with 8 mg/kg endostar administration daily, every other day or every two days. The peritoneal membrane permeability of the mice was assessed using Evan blue staining. The body weight curve of mice was drawn, and the cumulative ascites volume and number of red cells and tumor cells in the malignant ascites were determined. The survival of the mice was evaluated to assess the therapeutic effect of endostar.

RESULTSCompared with the control group, the mice receiving daily endostar injection showed obviously lower ascites accumulation and peritoneal capillary permeability (P<0.05) with reduced count of ascites tumor cells and red cells and tumor burden of the abdominal cavity. The mice with daily endostar injection also showed longer survival than the control group.

CONCLUSIONSContinuous intraperitoneal injection can be the optimal means for endostar administration for treatment of malignant ascites.

Animals ; Carcinoma, Ehrlich Tumor ; blood supply ; Cell Line, Tumor ; Endostatins ; administration & dosage ; pharmacology ; therapeutic use ; Female ; Humans ; Mice ; Mice, Inbred ICR ; Neovascularization, Pathologic ; Recombinant Proteins ; administration & dosage ; pharmacology ; therapeutic use

OBJECTIVETo investigate the role of Chengqi Shengxue prescription in anti-tumor and immunoregulation and to evaluate its effect on apoptosis and T lymphocyte subsets of tumor-bearing mice.

METHODS180 ascites tumor and Lewis lung carcinoma tumor-bearing mice were used in the screening. Then 55 mice were treated randomly with the model, cyclophosphamide (30 mg x kg(-1)), or three different dosages of Chengqi Shengxue prescription (2. 4, 1.2, 0.6 g x kg(-1). After the treatment apoptosis of tumor cell and peripheral T lymphocyte subsets of tumor-bearing mice was analyzed by flow cytometry.

RESULTLewis lung carcinoma was a nsitive tumor cell line to Chengqi Shengxue prescription. Compared with the model group, significantly increased apoptosis was observed after administration of high and medium dose of Chengqi Shengxue prescription (P < 0. 05) by PI staining. Increased early apoptosis in cancer cells was observed in all experimental doses of Chengqi Shengxue prescription by Annexin V and PI double staining (P < 0.01) . The analysis of T lymphocyte subsets showed that the percentage of CD3, CD4 and CD4/CD8 ratio decreased significantly in model group when compared with the normal ones (P < 0.01), while no change was observed in CD8. In administration groups, CD3, CD4 and CD8 were significantly lower than normal ones (P < 0.01) , but CD4/CD8 ratio did not change significantly.

CONCLUSIONChengqi Shengxue prescription has selectively inhibitive effect on the growth of mouse Lewis lung carcinoma and takes an antitransfer role. Its anti-tumor effect may be owing to inducing tumor cell apoptosis. Chengqi Shengxue prescription improves cellular immune function through enhancing CD4/CD8 ratio.

Animals ; Antineoplastic Agents ; therapeutic use ; Apoptosis ; drug effects ; Carcinoma, Ehrlich Tumor ; drug therapy ; Carcinoma, Lewis Lung ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Male ; Mice ; Mice, Inbred C57BL ; Neoplasms, Experimental ; drug therapy ; T-Lymphocyte Subsets ; drug effects ; immunology

OBJECTIVETo evaluate if 99mTc-HYNIC-annexin V may be used to detect the early chemotherapeutic effect and to determine the best timing for detecting apoptosis in vivo.

METHODSAnnexin V was labeled with 99mTc using HYNIC as a bifunctional agent. Normal Kunming mice received inoculation of Ehrlich ascites cells into the right upper limb. After the tumor reached 1 cm in diameter, the mice were randomly divided into saline treatment group as control and cyclophosphamide (150 mg/kg injected intraperitoneally) treatment group. 99mTc-HYNIC-annexin V was injected intravenously at 1 h and 24 h after treatment. Region of interest technique (ROI) from the SPECT images taken at different time was used to get the ratio of tumor/limb in each group. TUNEL staining was used to detect apoptotic cells and the rates of positive stained cells were calculated.

RESULTSAfter treatment with saline, only little amount of the radiolabeled tracer could be seen in the tumor and showed weak image of the tumor. But after 24 h of treatment with cyclophosphamide, clear image on the tumor could be seen. 24 h after the treatment of cyclophosphamide, the ratio of tumor/limb was (6.27 +/- 0.24) which was much higher than that at 24 h after treatment with saline (2.36 +/- 0.18) and that at 1 h after cyclophosphamide treatment (4.00 +/- 0.38). At 24 h after cyclophosphamide treatment, TUNEL staining showed a significantly higher rate of apoptotic cells in the mice.

CONCLUSION99mTc-HYNIC-annexin V can be used as an apoptosis-imaging agent to detect and evaluate the early curative effect after chemotherapy. The effective detection of apoptotic response in tumor with 99mTc-HYNIC-annexin V requires a 24 h interval after chemotherapy. SPECT images can be obtained at 60 min after injection of the imaging agent. It suggests that 99mTc-HYNIC-annexin V may become a promising agent for apoptosis-imaging in clinical application.

Animals ; Annexin A5 ; pharmacokinetics ; Antineoplastic Agents, Alkylating ; therapeutic use ; Apoptosis ; drug effects ; Carcinoma, Ehrlich Tumor ; diagnostic imaging ; drug therapy ; pathology ; Cyclophosphamide ; therapeutic use ; Male ; Mice ; Neoplasm Transplantation ; Organotechnetium Compounds ; pharmacokinetics ; Radiopharmaceuticals ; pharmacokinetics ; Random Allocation ; Tissue Distribution ; Tomography, Emission-Computed, Single-Photon

OBJECTIVETo investigate the immunoregulatory activities of polysaccharopeptide and astragalus polysaccharides on EAC tumor-bearing mice.

METHODEhrlich's ascites carcinoma (EAC) Kunming (KM) mice were used to establish the animal model for solid tumor. Mice were randomly divided into six groups (n = 10): NS group (NS, 10 mL x kg(-1) x d(-1)), AMD group (AMD, 4 mg x kg(-1) x d(-1), 0.2 mL, only for the first 3 days), PSP group (PSP, 250 mg x kg(-1) x d(-1), 0.2 mL), APS group (APS, 250 mg x kg(-1) x d(-1), 0.2 mL), complex prescription group (PSP + APS, 250 mg x kg(-1) x d(-1), 0.1 mL) and combined treat group (AMD + PSP + APS, same dosage as above). After thirty days of treatment, immunocytochemical method was employed to detect the changes of T-lymphocyte subsets in the PBMC of tumor-bearing mice. Subsequently, the organ indexes and tumor inhibition rate were calculated and compared with those of control group.

RESULTPercentage of CD3+, CD4+ T-cell and the ratio of CD4+/CD8+ were obviously prominence in the PSP and PSP + APS groups compared with those of NS group (0.05), percentage of CD8+ T-cell was significantly decreased compared with that of AMD group; percentage of CD3+, CD4+ T-cell were obviously increased in AMD + PSP + APS group relative to that of AMD group; the thymus index of AMD group was significantly decreased compared with that of NS group, but the thymus index of AMD + PSP + APS group was obviously increased compared with that of AMD group; the weight of tumor in each administration group was significantly decreased compared with that of NS group.

CONCLUSIONPSP and PSP + APS complex prescription showed the remarkable immunoregulation on EAC mice with chemotherapy or not.

Animals ; Antineoplastic Combined Chemotherapy Protocols ; Astragalus Plant ; chemistry ; immunology ; Carcinoma, Ehrlich Tumor ; drug therapy ; immunology ; pathology ; Female ; Mice ; Polysaccharides ; administration & dosage ; immunology ; pharmacology ; therapeutic use ; Proteoglycans ; administration & dosage ; immunology ; pharmacology ; therapeutic use ; T-Lymphocyte Subsets ; drug effects ; immunology

OBJECTIVETo investigate anticancer effects and potential mechanisms of CL, extract of Rosa roxburghii.

METHODIn vitro anticancer effect was observed in Ehrlich's ascites carcinoma (EAC) mice model. Cell toxicity of CL on human endometrial adenocarcinoma cell line JEC (JEC) cells was measured by MTT reduction test and growth curves drawing by trypan blue dye exclusion method. Lactate dehydrogenase (LDH) concentration of cultured medium was detected by auto-biochemistry-meter. Cell differentiation was showed by detection of NBT reduction ability. Apoptosis was showed by AO/EB fluorescent staining and flow cytometer detection. Cell proliferation cycle was detected by flow cytometer.

RESULTComparing with the negative group, life span of EAC mice treated with CL was prolonged (P <0.05), and thymus index and spleen index of them were raised (P <0.05). The inhibitory effect of CL on JEC cells was in concentration-and time-dependent manner. IC50 of CL on JEC cells was 0.05 microg mL(-1) in 96 hours. Growth curves showed right-shift with CL concentration increasing. The number of NBT positive JEC cells increased and the LDH concentration of cultured medium declined with CL increasing. Apoptosis of JEC cells with CL treated was induced in concentration-dependent manner, apoptotic percentage of CL 10 microg mL(-1) on JEC cells was 25.59% in 24 hours. CL arrested JEC cells in G2-M phase (P <0.05).

CONCLUSIONCL has certainly anticancer effects in vivo and in vitro. Anticancer effect of CL in vivo was in relation to enhancing immune function of EAC mice; anticancer mechanisms of CL on JEC cells may be its direct cytotoxic effect, inducing cell apoptosis and inhibiting cell segmentation.

Adenocarcinoma ; metabolism ; pathology ; Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Ehrlich Tumor ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Endometrial Neoplasms ; metabolism ; pathology ; Female ; Humans ; L-Lactate Dehydrogenase ; metabolism ; Mice ; Plants, Medicinal ; chemistry ; Random Allocation ; Rosa ; chemistry