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整合素αVβ3,脂筏和c-SRC在人类黑色素瘤A375细胞迁移中的作用研究

摘要第4-5页
ABSTRACT第5页
CONTENTS第6-9页
INTRODUCTION第9-32页
    0.1 GLOBAL CANCER STATISTICS第14-15页
    0.2 CONTROVERSIES OVER THE EXISTENCE OF LIPID RAFTS第15-16页
    0.3 The STRUCTURAL ANALYSIS AND COMPOSITION OF LIPID RAFTS, INTEGRIN αVβ3 AND c-SRC第16-22页
        0.3.1 Lipid Rafts第16-18页
        0.3.2 Integrins第18-20页
        0.3.3 The structural composition and activation of c-SRC第20-22页
    0.4THE ROLE OF LIPID RAFTS, c-SRC and INTEGRIN αVβ3 IN REGULATING CANCER CELL MIGRATION第22-25页
        0.4.1 Lipid Rafts Regulate Cell Migration Through Enhancing Signal Transduction第22-23页
        0.4.2 Integrins'Adhesive Structures Form Linkage Between The ECM And Actin Cytoskeleton To Bring About Migration第23-24页
        0.4.3 c-SRC Acts As A Kinase To Regulate Migration第24-25页
    0.5 THE CROSSTALK BETWEEN LIPID RAFTS, c-SRC AND INTEGRIN αVβ3第25-28页
    0.6 MODELS USE IN THE STUDY OF CELL MIGRATION第28-29页
    0.7 RESEARCH AIMS第29-30页
    0.8 RESEARCH SIGNIFICANCE第30-32页
MATERIALS第32-35页
    1.1 TUMOR CELL LINE第32页
    1.2 COMPETENT CELLS STRAIN第32页
    1.3 ANTIBODIES第32页
    1.4 REAGENTS第32-33页
    1.5 MAIN INSTRUMENTS第33页
    1.6 MEDIA AND SOLUTION PREPARATIONS第33-35页
        1.6.1 SDS-PAGE, Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis Solution第34页
        1.6.2 Western Blot Buffers第34-35页
        1.6.3 LB (Luria Bertani) Medium Preparation第35页
        1.6.4 Agarose Gel Preparation第35页
METHODS第35-51页
    2.1 CELL CULTURE第35页
    2.2 c-SRC DOMAINS PLASMID CONSTRUCT第35-42页
        2.2.1 Extracting the plasmid from the filter paper第36页
        2.2.2 Transformation第36页
        2.2.3 Culturing and storing single clones第36页
        2.2.4 Extraction of plasmid from grown cultured medium第36-38页
        2.2.5 Polymerase Chain Reaction(PCR)第38-39页
        2.2.6 PCR Master Mix第39页
        2.2.7 Primers第39-40页
        2.2.8 Running the gel第40页
        2.2.9 Purifying the gel第40-41页
        2.2.10 Restriction cutting第41页
        2.2.11 Ligation第41-42页
    2.3 CELLS TRANSFECTION第42页
    2.4 siRNA INTEGRIN β3 PREPARATION第42-45页
        2.4.1 Making cDNA第43-44页
        2.4.2 PCR Master Mix第44页
        2.4.3 PCR Cycle Reaction Times第44页
        2.4.4 Integrin β3 Primers第44页
        2.4.5 Confocal Immunofluorescence Microscopy第44-45页
    2.5 MTT CELL PROLIFERATION ASSAY第45页
    2.6 WOUND HEALING ASSAY第45-46页
        2.6.1 Calculations第46页
    2.7 TRANSWELL ASSAY第46-47页
    2.8 COIMMUNOPRECIPITATION AND WESTERN BLOT第47-48页
        2.8.1 Western Blot第47-48页
    2.9 CONFOCAL IMMUNOFLUORESCENCE MICROSCOPY FOR COLOCALIZATION EXPERIMENT第48页
    2.10 STATISTICAL ANALYSIS第48-51页
RESULTS第51-62页
    3.0 LIPID RAFTS AND c-SRC AFFECTS CELL PROLIFERATION第51-52页
    3.1 LIPID RAFT DISRUPTION AND c-SRC INHIBITION INDEPENDENTLY REDUCES A375 CELL MIGRATION第52-57页
    3.2 OVEREXPRESSION OF c-SRC DOES NOT INCREASE A375 CELLS MIGRATION第57-58页
    3.3 DEPLETION OF INTEGRIN β3 CAUSES A DECREASE IN A375 CELL MIGRATION第58-60页
    3.4 LIPID RAFT DISRUPTION DOES NOT AFFECT c-SRC INTERACTION WITH INTEGRIN αVβ3第60-62页
DISCUSSION第62-65页
CONCLUSION第65-66页
REFERENCES第66-73页
ACKNOWLEDGEMENT第73-74页
PUBLISHED PAPERS DURING MASTER DEGREE (2010-2013)第74页

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