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Mitochondrial shape rapidly changes by dynamic balance of fusion and fission to adjust to constantly changing energy demands of cancer cells. Mitochondrial dynamics balance is exactly regulated by molecular motor consisted of myosin and actin cytoskeleton proteins. Thus, targeting myosin-actin molecular motor is considered as a promising strategy for anti-cancer. In this study, we performed a proof-of-concept study with a natural-derived small-molecule J13 to test the feasibility of anti-cancer therapeutics
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Objective To establish model of the chicken embryo transplantation of human colon cancer cells ,and investigate the effect of Solanine、VEGF antibody and Solanine combined with VEGF antibody on human colon cancer cells induce tumor angio‐genesis and tumor proliferation .Methods The model of the chicken embryo transplantation of human colon cancer HT‐29 cells were divided into three experimental group and control group .We added to the chick embryo chorioallantoic membrane with Sola‐nine、VEGF antibody and Solanine+ VEGF antibody mixture ,PBS was added to the control group .Then we analysed picture through the stereomicroscope and IPP 6 .0 image analysis software ,using immunohistochemistry envision method to detect of CD34 antigen and ki‐67 antigen ,and observing effect of Solanine group ,VEGF antibody group ,Solanine+ VEGF antibody group and the effect on the tumor angiogenesis and tumor proliferation .Results The tumor angiogenesis ,CD34 antigen and ki‐67 antigen of Sola‐nine+VEGF antibody group were significantly better than those of VEGF antibody group and Solanine group(P<0 .01);VEGF antibody group had statistical significant difference with Solanine group(P<0 .01);the effect of other three groups were better than that of the control group(P<0 .01) .Conclusion Solanine、VEGF antibody and Solanine combined with VEGF antibody could in‐hibit tumor angiogenesis and tumor proliferation of human colon cancer cell line HT‐29 to induce .It provides a new way for anti‐an‐giogenes .
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The P1.HTR cell line includes highly transfectable cells derived from P815 mastocytoma cells originating from mouse breast tissue. Despite its widespread use in immunogenic studies, no data are available about the behavior of P1.HTR cells in the chick embryo chorioallantoic membrane model. The objective of the present investigation was to study the effects of P1.HTR cells implanted on the chorioallantoic membrane of chick embryos. We inoculated P1.HTR cells into the previously prepared chick embryo chorioallantoic membrane and observed the early and late effects of these cells by stereomicroscopy, histochemistry and immunohistochemistry. A highly angiotropic and angiogenic effect occurred early after inoculation and a tumorigenic potential with the development of mastocytoma keeping well mast cells immunophenotype was detected later during the development. The P1.HTR mastocytoma cell line is a good tool for the development of the chick embryo chorioallantoic membrane mastocytoma model and also for other studies concerning the involvement of blood vessels. The chick embryo chorioallantoic membrane model of mastocytoma retains the mast cell immunophenotype under experimental conditions and could be used as an experimental tool for in vivo preliminary testing of antitumor and antivascular drugs.
Subject(s)
Animals , Chick Embryo , Chorioallantoic Membrane/pathology , Mastocytoma/pathology , Cell Line, Tumor , Chorioallantoic Membrane/blood supply , Immunohistochemistry , Neovascularization, PathologicABSTRACT
Objective To optimize the conditions for enhancing the refolding of a novel recombinant human(rh)endostatin and test the biological activities of refolded endostatin.Methods The partial purified inclusion bodies of rh-endostatin were dissolved with 6 mol/L guanidine-HCl followed by combination of dilution and dialysis of the dissolved endostatin.The refolded endostatin was then purified by cation-exchange chromatography.The biological activities of purified rh-endostatin were assessed by endostatin-specific monoclonal antibody and chick embryo chorioallantoic membrane assay.Results A 46% refolding yield was achieved after optimizing the refolding conditions.The purified endostatin reacted with specific anti-endostatin monoclonal antibody and showed significant inhibition of angiogenesis in chick embryo ehorioallantoic membrane assay.Conclusion The method of highest refolding yield of human endostatin was developed.This optimized method significantly promotes the application of this novel human endostatin to preclinical and clinical studies.
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The effects of Friend erythroleukemia cells on angiogenesis were studied in chick embryo chorioallantoic membrane assay and in human umbilical vein endothelial cells. In chorioallantoic membrane assay, the conditioned medium of Friend cells stimulated in vivo angiogenesis to an extent comparable to that observed with Prostaglandin El, used as positive control. Prostaglandin El added to conditioned medium of Friend cells did not further increase angiogenesis. Conditioned medium of Friend erythroleukemia cells also stimulated proliferation of human umbilical vein endothelial cells to an extent comparable to that observed with fetal bovine serum, used as positive control. Conditioned medium and fetal bovine serum together did not affect human umbilical vein endothelial cells proliferation, as compared to that observed when tested separately. These results seem to indicate that Friend erythroleukemia cells produce and secrete factors stimulating angiogenesis. These findings extend and confirm the hypothesis that successful angiogenesis is necessary for development of leukemias.
Subject(s)
Animals , Cattle , Chick Embryo , Humans , Chorioallantoic Membrane/blood supply , Friend murine leukemia virus , Leukemia, Erythroblastic, Acute/pathology , Neovascularization, Pathologic/etiology , Umbilical Veins/cytology , Cell Proliferation , Endothelial Cells/pathology , Leukemia, Erythroblastic, Acute/metabolism , Tumor Cells, CulturedABSTRACT
In order to investigate the angiogenic effect of intercellular adhesion molecule-1 (ICAM-1), two parts of experiment were performed. Chick embryo chorioallantoic membrane (CAM) assay was used for in vivo angiogenic research. The chick embryos were divided into 4 groups: ICAM-1 group (divided into 3 subgroups, Ⅰ, Ⅱ and Ⅲ) for screening the angiogenic effect of ICAM-1 by adding different concentrations of ICAM-1 (0.1, 0.2 and 0.3 μg/μL) 5 μL into the chick embryo CAMs on the day 10 after incubation for every subgroup; Anti-ICAM-1 group A (divided into 2 subgroups, Ⅰ and Ⅱ) by adding different concentrations of Anti-ICAM-1 (1:100, 1:50) 5 μL into the chick embryo CAMs on the day 10 after incubation for every subgroup to evaluate the effect of ICAM-1 on the survival of microvessels through observing whether Anti-ICAM-1 could induce involution of the microvessels on CAMs; Anti-ICAM-1 group B (divided into 2 subgroups, Ⅰ and Ⅱ ) by adding different concentrations of Anti-ICAM-1 (1:100, 1:50) 5 μL into the chick embryo CAMs on the day 6 after incubation for every subgroup to evaluate whether ICAM-1 involved in embryonic angiogenesis through observing the growth of microvessels on CAMs; Control group: ICAM-1 or Anti-ICAM-1 was substituted by PBS 5 μL on the day 10 or day 6 after incubation. Three days later, the CAMs were photographed in vivo, excised, sectioned and the number of microvessels was counted. In ICAM-1 group, there was increased number of microvessels arranged radially with "spoked-wheel" pattern around the gelatin sponges. The new microvessels growing perpendicularly to gelatin sponges were observed. The number of the microvessels growing in the CAM mesenchymes around the sponges in 3 subgroups was higher than that in control group (P＜0.01), however, there was no significant difference among the 3 subgroups (P＞0.05). In anti-ICAM-1 group A, the radially arranged microvessels were very unclear around the sponges contrast to that of ICAM-1 group. Few new microvessels were detected in the center of the sponges. The number of the microvessels growing in the CAM mesenchymes around the sponges in subgroup Ⅱ was lower than that in control group (P＜0.01). There was no significant difference in the number of the microvessels around the sponges between subgroup Ⅰ and control group (P＞0.05). In anti-ICAM-1 group B, the radially arranged microvessels were very unclear around the sponges contrast to that of control group. New microvessels were very scarce in the center of the sponges. The number of the microvessels growing in the CAM mesenchymes around the sponges in the 2 subgroups were less than that in control group (P＜0.01), and there was significant difference between the 2 subgroups (P＜0.05). It was suggested that ICAM-1 could induce angiogenesis and support the survival of microvessels, and ICAM-1 was involved in embryonic angiogenesis.
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Background and purpose:Norcantharidin(NCTD)can inhibit the growth of tumors.In this study,we presented the chorioallantoci membrane(CAM)model to evaluate the effect of NTCD on angiogenesis and to study the anti-angiogenesis of NTCD in the chicken embro implant model of human breast carcinoma MCF-7 cells.Methods:The method was based on the implantation of gelatin sponges on top of growing CAM.The sponges were treated with various amounts of NCTD and 0.9% NaCl.Blood vessels surrounding CAM mesenchyme were counted.Then we established the MCF-7 chicken embryo implant model.The chicken embryo CAMs was treated with 0.9% NaCl,Various amounts of NCTD(72,36,18 ?g per 20 ?l)and the inhibition rates were calculated.Results:NCTD can inhibit the new capillary vessels growing into gelatin sponges placed on the CAM in a dose-dependent manner,and the inhibitory rates were 77.7%,62.9%,50.6% and 33.0% respectively.NCTD at the dosages of 72、36、18 ?g were able to inhibit capillary growth regardless of the angiogenic process induced by xenograft tumor and had a significant inhibition as compared with the control group,and the rates were 66.2%,39.3%,22.8% respectively.Conclusions:NCTD has an anti-angiogenic effect for targeting tumor angiogenesis.NCTD may be a potential therapeutic candidate for clinical application.
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Objective:To study the influence of Sinapine on the formation of microvessels in chick embryo chorioallantoic membrane(CAM).Methods: Sinapine was purified by the procedure of pressurized-solvent extraction from Semen Sinapis Albae.The inhibition effect of Sinapine on the chorioallantoic vessel was detected by the chick embryo chorioallantoic membrane model.Results: Compared with physiological saline group,large areas of blood vessels in Sinapine groups(5.0g/L and 10.0g/L) had heavy inhibition of angiogenesis on CAM,accompanied by the decreased density of vessels(P0.05).Conclusions: Sinapine can prevent the formation of microvessels of CAM and the effects depend on the concentration.
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Purpose:To study the characterics of angiogenesis induced by osteosarcoma OS-732 cell line. Methods:With a tumor model of the chick embryo chorioallantoic membrane(CAM),the angiogenesis induced by OS-732 cell line was observed by a stereo-microscope and transmission electron microscope, and the expression of angiogenic factors vascular endothelial growth factor(VEGF) and basic fibroblast growth factor(bFGF) in xenografts of OS-732 cell line on CAM was detected by immunohistochemical technique. Results:This cell line was strongly angiogenic, which probably is associated with production of many factors.The new capillaries converging upon the tumor can be observed by a stereo-microscope. Furthermore,under transmission electron microscope, it also can be noted that the new blood vessel consisted of a monolayer of vascular endothelial cells with enlarged fissures,and the basal membrane was not intact. The expression of VEGF and bFGF was positive in xenografts on CAM, and VEGF was expressed at high levels constantly. Conclusions:The new blood vessels induced by tumors have pathologic and physiopathologic characteristics and many angiogenic factors are alm involved in the angiogenic process of OS-732. Furthermore, the application of angiogenic factors as a target in antiangiogenic therapy of osteosarcoma may be useful to improve the prognosis of patients with this disease.