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@#Objective: To investigate the expression of Artemin in chondrosarcoma and its effect on proliferation and migration of endothelial cells, and to explore the mechanism. Methods: A total of 40 chondrosarcoma tissue samples (low degree (Ⅰ), 20 cases; high degree (Ⅱ,Ⅲ), 20 cases) surgically resected from patients, who were treated in Lianyungang HospitalAffiliated to Nanjing University of Chinese Medicine from May, 2015 to April, 2019, were collected for this study. Another 20 cases of normal cartilage tissue specimen from patients with amputations due to car accidents were served as control. The expressions of Artemin, vascular endothelial growth factor (VEGF), Ki-67 and CD31+ vascular density in tumor tissues were detected by immunohistochemistry.After being treated with 10 ng/ml Artemin, the changes of VEGF, stromal cell derived factor-1 (SDF-1), matric metalloproteinase 2 (MMP2) and MMP9 in the supernatant of SW1353 cell culture were detected by enzyme-linked immunosorbent assay (ELISA), and the effects of Artemin-treated chondrosarcoma cells on the migration and proliferation of ECV304 cells were detected by Transwell migration assay and MTT cell proliferation assay, respectively. Results: The expressions of Artemin and Ki-67 in the tissues of low-level group were significantly higher than those in the control group (all P<0.01); the expressions ofArtemin and Ki-67 in the tissues of high-level group were significantly higher than those in the low-level group (all P<0.01). The expression of Artemin was positively correlated with VEGF level and vascular density in chondrosarcoma tissues (all P<0.01);Artemin promoted the secretion of VEGF by chondrosarcoma cells, but had no significant effect on the secretion of SDF-1, MMP2 and MMP9. Artemin induced the proliferation and migration of ECV304 cells by promoting the secretion of VEGF by chondrosarcoma cells (all P<0.01). Conclusion: Artemin is highly expressed in chondrosarcoma tissues and has a positive correlation with the expression of VEGF and vascular density. Artemin can enhance the angiogenesis induced by chondrosarcoma.
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Aim To investigate the effect of MTX(included(±)MTX,(+)MTX and(-)MTX)on the proliferation of ECV304 cells and to explore its mechanisms.Methods ECV304 cells were cultured.The cell proliferation was determined by MTT.The morphological changes were inspected by inverted microscope.Cell cycle phases were assayed by propidium iodide staining flow cytometry.Results ECV304 cells were treated with(+)MTX,(-)MTX and(±)MTX at 1～150 μmol·L~(-1) for 24,48,72 h.The results showed that the proliferation of ECV304 cells was significantly inhibited under different conditions.The order of the inhibited efficacy was(+)MTX>(±)MTX>(-)MTX.The morphology of ECV304 cells were changed by(+)MTX,(-)MTX and(±)MTX treatment,which included the cell shrinkage,chromatin condensation.After administration of 10 μmol·L~(-1) of(+)MTX,(-)MTX and(±)MTX for 48 h,the cell cycle phases were assayed by propidium iodide staining flow cytometry.The result showed DNA replication was interfered by(+)MTX,(-)MTX and(±)MTX treatment.Conclusions The proliferation of ECV304 cells has the chiral selective effects by(+)MTX and(-)MTX treatment,and the inhibition on ECV304 cells proliferation of(+)MTX is significantly stronger than that of (-)MTX.
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Objective To study the effect of the ultra-filtration extract from Angelica sinensis and Hedysarum polybotrys’s mixture on the VEGF mRNA expression of ECV-304 cell. Method Angelica sinensis and Hedysarum polybotrys’s mixture were refined by ultra-filtration technique. ECV-304 cells were cultured as model, and its proliferation was detected by MTT colorimetry. VEGF mRNA expression was observed by semi-quantitative RT-PCR. Results The ultra-filtration extract from Angelica sinensis and Hedysarum polybotrys’s mixture could markly promote the growth of ECV-304 cells, there was significant difference between experimental and control group (P
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Objective:To develop an experimental three-dimensional model by ECV304 cells(human umbilical vein endothelial cell line) for investigating the mechanisms of angiogenesis in vitro.Methods:ECV304 cells were seeded onto three-dimensional collagen gels made of rat-tail collagen.When the endothelial cells were cultured and grown to near confluence,treated with bFGF for 3 to 12 days,and then assessed with inverted phase contrast microscope.Results:The endothelial cells migrated into the gels,formed complex networks by cell cords at different levels through the bottom view,and sprouted capillary-like structures through the side view.Conclusion:ECV304 cells are capable of expressing some early events of angiogenesis in the three-dimensional collagen gels:proliferating,migrating and sprouting and so on.It should be useful for studying angiogenesis in vitro.