Dissertation > Excellent graduate degree dissertation topics show

The Inhibitory Effects of Celastrol on the Process of Brain Glioma Stem Cells’ Differentiation into Vascular Endothelial Cell-like Cells

Author: CaoGuangHui
Tutor: ZhouYouXin
School: Suzhou University
Course: Neurosurgery
Keywords: brain glioma stem cells differentiation vascular endothelial cell-like cells celastrol cell apoptosis cell cycle
CLC: R739.4
Type: Master's thesis
Year: 2011
Downloads: 48
Quote: 0
Read: Download Dissertation

Abstract


At present, the brain glioma stem cells theory has been more and more accepted by many scholars. Brain glioma stem cells （BGSCs） are seed cells of glioma,having stem cells’attributes such as self-renew, proliferation and multiple differentiation potential. In recent years, the researches have focused on glioma stem cells’differentiation into their parental phenotype cells called glioma cells both at home and abroad , but the studies on whether BGSCs cells can differentiate into vascular endothelial cell-like cells and then participate in tumor blood supply were less, so we studied on BGSCs’differentiation into vascular endothelial cell-like cells on certain conditions in vitro and application of celastrol inhibiting its differentiation process.The study involved two aspects of the glioma stem cells research:①brain glioma stem cells’differentiation into vascular endothelial cell-like cells on certain conditions in vitro.②the inhibitory effects of different concentrations celastrol on the differentiation process.PartⅠ:Brain glioma stem cells’differentiation into vascular endothelial cell-like cellsObjective To explore the feasibility of brain glioma stem cells’differention into vascular endothelial cells on certain conditions in vitro, and establish foundation for the further study of celastrol’s inhibitory effect of the differentiation process.Methods Brain glioma stem cells were cultured in neural stem cell culture solution , then cells’growth changes were observed and CD133, Nestin ,CD31 and KDR immunofluorescence stainings were used to examine the cells after passage. Brain glioma stem cells were divided into the experimental group and the control group after passage. Cells of the experimental group were cultured by differentiation culture solution which included basal culture solution DMEM/F12 100ml, 10%FCS, VEGF 20ng/ml, EGF 20ng/ml , bFGF 20ng/ml, 1/100 N2 in anaerobic environment which included 0%O2,5%CO2,10%H2 and 85%N2, while the cells of the control group were cultured by ordinary culture solution which included basal culture solution DMEM/F12 100ml, 10%FCS in aerobic environment. Cells’growth changes were observed by the microscope. After 12 days, CD31 and KDR immunofluorescence stainings were used to examine the cells, and the cells were planted on Matrigel glue to observe the cells’growth changes.Result Brain glioma stem cells suspended in neural stem cell culture solution and the results of CD133 and Nestin immunofluorescence stainings were positive while the results of CD31 and KDR immunofluorescence stainings were negative. The experimental group cells were cultured by differentiation culture solution in anaerobic environment for 12 days. Flag stone phenomenon was observed. Then the results of CD31 and KDR immunofluorescence stainings were positive and tubular structures were observed after the cells were planted on Matrigel glue for 24 hours. While the flag stone phenomenon was not observed in the control group cells cultured by ordinary culture solution in aerobic environment for 12 days.And the results of CD31 and KDR immunofluorescence stainings were negative and tubular structures were not observed after the cells were planted on Matrigel glue for 24 hours.Conclusion The brain glioma stem cells which were cultured by differentiation culture solution in anaerobic environment in vitro for 12 days could differentiate into vascular endothelial cell-like cells.PartⅡ:The inhibitory effects of celastrol on the process of the brain glioma stem cells’differentiation into vascular endothelial cell-like cellsObjective To study the inhibitory effects of different concentration celastrols on brain glioma stem cells and the effects and influences on the process of the brain glioma stem cells’differentiation into vascular endothelial cell-like cells.Methods One part of brain glioma stem cells differentiated for 72 hours. And then celastrols of different concentrations such as 0.04μg/ml、0.2μg/ml、1.0μg/ml、5.0μg/ml、25.0μg/ml、125.0μg/ml were added to culture the cells for 48 hours. Cells continued to differentiate for 7 days.Then cells’growth changes were observed and CD31, KDR immunofluorescence stainings were used. The other part of brain glioma stem cells were planted on the Matrigel glue, and celastrols of different concentrations were added like the front steps. We observed whether tubular structures could form. In addition, the effects of celastrol on the proliferation of glioma stem cells were examined by MTT assay after celastrols were added for respectively 24 and 48 hours in the process of differentiation. Cells were collected after the different concentration celastrols took effects for 48 hours; Cell apoptosis was examined by AnnexinⅤ-PI assay; Cell cycle changes were examined by the cell cycle kit; And caspase-3 expression was examined by Semi-quantitative RT - PCR method.Result Only in 0.04μg/ml group, the flag stone phenomenon appeared, CD31, KDR immunofluorescence stainings were positive,and tubular structures could form on the Matrigel glue.Few tubular structures could be observed in 0.2μg/ml group cells.While the results were negative in other group cells. With the increase of celastrol concentration, the inhibitory effects on cells’growth were more stronger by MTT assay, cell apoptosis was more obvious by AnnexinⅤ- PI method;And caspase-3 expression was more obvious by semi-quantitative RT - PCR method.Conclusion The celastrol could inhibit the growth of the brain glioma stem cells and the process of the brain glioma stem cells’differentiation into vascular endothelial cell-like cells significantly in certain concentration range .The inhibitory process was in time-dose dependence. The mechanisms why celastrol could inhibit the process of brain glioma stem cells’differentiation into vascular endothelial cells probably may relate with the facts that celastrol could introduce cells apoptosis and block cells in S period.

Related Dissertations

  1. Study of Oridonin on SGC-7901 Cell Proliferation by Inhibitting Cell Cycle Proteins,R285
  2. Exoression of Resistin During Porcine Preadipocyte Differentiation and Its Infiuencing Factors,R587.1
  3. Expression of Prolyl Isomerase Pin1 in Osteosarcoma and the Effect of Regulation on Cell Cycle,R738.1
  4. Professor Wang Qi identified the body - of Diseases - dialectical combination of academic thought and clinical experience and treatment of chronic insomnia clinical studies,R249.2
  5. miRNA-23a into the regulation of muscle differentiation in C2C12 cells,Q952.5
  6. miR - 23 in mouse skeletal muscle expression and the regulation of skeletal muscle regeneration research,Q78
  7. Evaluation of Land Use Eco-security in Fujian Province,X826
  8. Effects of BMPR-IB Gene Silencing by Small Interfering RNA on Apoptosis of Porcine Pollicular Granulosa Cells and Ecpression of BMP Pathy-Way-Ralted Genes,S828
  9. The Effect of Foxol on Apoptosis of Mouse Granule Cells,S865.13
  10. Heme Oxygenase is Involved in Salinity-Induced Tomato Lateral Root Development and Its Relationship with Hydrogen Peroxide,S641.2
  11. Identification of Biological Characteristics with Human Keloid Fibroblast-like Mesenchymal Stem Cells,R622
  12. The Involvement of Heme Oxygenase/Carbon Monoxide System in Cobalt Chloride-Induced Lateral Root Development in Tomato Seedlings,S641.2
  13. The Impact of Histone Deacetylase Inhibitor Trichostatin a on Proliferation and Apoptosis of Porcine Granulosa Cells,S828
  14. Study on the Cell Cycle of Liver Cells during Rat Liver Regeneration,Q253
  15. Effects of Different Immunization on the Development of Immune Organs,cell Proliferation and Expression of Apoptosis-relatived Gene in Chicken,S858.31
  16. Effects of Ginsenoside Rg1 on Cell Proliferation, Secretion Function and ERK、p38 Signal Molecular in Human Umbilical Cord Blood Mesenchymal Stem Cells under Optimized Culture Condition,R285.5
  17. Effects on Recombinant Human Interleukin-10 of IL-2, IFN-γ, GCs, T Cell Apoptosis in Transplantation of the Rabbit Skin,R392
  18. The Mechanism of CREG Regulated Vasculogenesis in Murine Embryonic Stem Cells,R541.4
  19. Ginsenoside Rg1 on human umbilical cord blood CD34 ~ hematopoietic stem cell proliferation and differentiation , and the impact of calcium-sensing receptor,R285.5
  20. VEGF-induced neuronal differentiation into mesenchymal stem cells migration research,R329
  21. Eastern China in the New Comparative Study of differentiation of the peasantry,D663.2

CLC: > Medicine, health > Oncology > Nervous system tumors
© 2012 www.DissertationTopic.Net  Mobile