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研究生： 李倫維
Lee, Lun-Wei
論文名稱： 探討循環拉伸誘導細胞柱狀化的機械生物學機制
Mechanobiological mechanism of cyclic stretch-induced cell columnarization
指導教授： 湯銘哲
Tang, Ming-Jer
學位類別： 博士
Doctor
系所名稱： 醫學院 - 基礎醫學研究所
Institute of Basic Medical Sciences
論文出版年： 2024
畢業學年度： 112
語文別： 英文
論文頁數： 111
中文關鍵詞： 上皮細胞柱狀化 、細胞骨架重組 、細胞力學 、循環拉伸 、機械力生物學
外文關鍵詞： Epithelial cell columnarization, Cytoskeleton rearrangement, Cellular mechanics, Cyclic stretch, Mechanobiology
相關次數： 點閱：32  下載：0 
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    • 中文摘要 I Abstract II 致謝 III Contents IV List of Tables VIII List of Figures IX Abbreviations XII Chapter 1: Introduction 1 1-1 Epithelial cell shape and mechanical stimuli 1 1-1.1 The shape of epithelial cells and functions 1 1-1.2 Mechanical stimuli: The intrinsic forces 2 1-1.3 Mechanical stimuli: The extrinsic forces 3 1-1.4 The correlation between epithelial morphogenesis and mechanical stimulation 3 1-2 Cytoskeletons in epithelial cell shape, polarity, and junction integrity 4 1-2.1 Actin filaments 5 1-2.2 Microtubules 6 1-3 Cell-cell and cell-matrix adhesions in epithelial integrity and mechanosensing 7 1-3.1 Tight junctions 8 1-3.2 Adherens junctions 9 1-3.3 Focal adhesion complex 10 1-4 Mechano-adaptation in epithelial cell behavior and tissue homeostasis 10 1-4.1 The importance of mechano-adaptation 10 1-4.2 Loss of mechanosensing and pathological conditions 11 1-5 Investigating long-term dynamic mechanical stimulation on epithelial morphogenesis 12 1-5.1 Methods for evaluating the effects of mechanical stimulations 12 1-5.2 Application of uniaxial cyclic stretching 13 1-5.3 Objectives 13 Chapter 2: Specific aims 15 Specific aim 1: To verify the geometrical features of columnarized cell shape under CS 15 Specific aim 2: To investigate the mechanism in promoting CS-induced cell columnarization 16 Specific aim 3: To investigate the change of cell mechanics in CS-induced columnarized epithelial cells 17 Chapter 3: Materials and methods 19 3-1 Cell lines and pharmacological treatment 19 3-2 Application of cyclic stretch 20 3-3 Western blotting 20 3-4 Transfection and knockdown 21 3-5 Immunofluorescence staining and microscopy 21 3-6 Cell shape analysis 22 3-7 Tortuosity analysis 22 3-8 Immunofluorescent intensity analysis 23 3-9 Analysis of apical surface stiffness by confocal-coaxial atomic force microscopy (AFM) 23 3-10 Modeling of epithelial cell columnarization 24 3-11 RNA sequencing analysis 26 3-12 Real-time cell analysis (RTCA) 27 3-13 Statistical analysis 27 Chapter 4: Results 28 4-1 Uniaxial cyclic stretch induces columnarization in MDCK epithelial cells 28 4-2 CS-induced cell columnarization is not dependent on cell proliferation and cell density 29 4-3 CS induced actin roof formation and thickening of microtubule network at the apical part of columnarized MDCK cells 30 4-4 Perturbation of cell force transmission ameliorated CS-induced cell columnarization 31 4-5 Inhibition of ZO-1 and actomyosin contractility alleviate CS-induced cell columnarization 32 4-6 Caveolin-1 overexpression augmented CS-induced cell columnarization by strengthening apical junction 34 4-7 Alteration of apical junctional stiffness and myosin contraction activity during cyclic stretch 36 4-8 Mathematical modeling depicts the force changes in CS-induced cell columnarization 37 4-9 Screening the possible signaling pathways that determine CS-induced cell columnarization by using RNA sequencing 39 Chapter 5: Discussion 41 5-1 The role of apical tension and basal tension in CS-induced columnar cell shape development 41 5-2 Synergistic effects of Cav1 overexpression and CS on cell height and columnar morphology 41 5-3 The impact of CS on cell density, proliferation and columnarization 42 5-4 Nuclear reorientation and the role of the LINC complex in CS-induced columnarization 43 5-5 The role of Mechano-adaptation in CS-induced columnarization and its implications 43 5-6 The regulation of focal adhesion complex in CS-induced cell columnarization 44 5-7 The implications of vinculin translocation on cell columnarization 45 5-8 The role of Ecad in promoting columnar shape formation in Cav1-overexpressing MK4 cells 46 5-9 The postulated mechanism of CS-induced cell columnarization 46 Conclusion 48 References 49 Figures 59 Tables 93

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