干细胞分裂模式在前列腺发育和肿瘤发生中的机制研究

发布时间：2018-01-27 13:08

本文关键词： 组织干细胞细胞分裂模式细胞命运发生前列腺发育前列腺肿瘤　出处：《上海交通大学》2015年博士论文　论文类型：学位论文

【摘要】：背景:干细胞对称和不对称分裂在组织发育,再生以及肿瘤发生发展中起到重要作用。虽然干细胞分裂模式的调控机理在模式动物果蝇和线虫以及哺乳动物的神经组织中得到广泛研究,但在哺乳类非神经组织或器官中,干细胞的分裂模式还很少报道。本论文中,我们通过细胞命运分析和细胞谱系追踪技术系统地研究前列腺正常发育及肿瘤发生过程中基底细胞和腔细胞的分裂模式。目的:(1)阐明前列腺正常发育以及再生过程中的基底细胞和腔细胞极性方式、细胞纺锤体定位和细胞命运决定等关键问题。(2)通过细胞谱系示踪以及基底细胞敲除小鼠研究基底细胞和腔细胞的命运发生。(3)研究Pten缺失是否通过影响基底细胞和腔细胞的细胞极性,分裂轴以及命运决定而促进前列腺肿瘤发生。材料和方法:(1)我们利用多种前列腺上皮基底细胞和腔细胞标记物的特异性抗体,细胞分裂标志物的抗体以及细胞极性的特异性抗体鉴定了基底细胞和腔细胞的极性方式以及纺锤体定位模式。(2)本研究使用多种转基因小鼠,包括前列腺基底细胞和腔细胞的谱系追踪小鼠,基底细胞谱系敲除小鼠以及前列腺特异性Pten敲除小鼠。(3)我们通过正置荧光显微镜和激光共聚焦显微镜捕捉基底细胞和腔细胞细胞命运变化。结果:第一章中,我们观察到基底细胞和腔细胞具有不同的细胞极性方式以及纺锤体定位模式。第二章中,通过细胞命运的分析证明基底细胞的水平分裂为对称式扩增分裂,而垂直分裂则为不对称分裂,产生分化的腔细胞。相比较,腔细胞只进行对称分裂,产生的子细胞都为腔细胞。谱系追踪实验进一步验证基底干细胞的双潜能性以及腔细胞的单潜能性。第三章中,我们证明在敲除Pten后,腔细胞自身的对称分裂能加速腔细胞群的扩增并使前列腺癌呈现腔表型,而转化的基底细胞仍然可进行不对称分裂产生腔细胞并诱导产生腔细胞样的前列腺肿瘤。第四章中,我们发现表达Sox2以及Trop2的腔细胞在前列腺肿瘤发生过程中都增多,通过谱系分析以及细胞分裂模式分析,证明腔细胞在谱系发生上可由基底细胞生成。结论:通过对前列腺上皮细胞分裂模式的研究,我们发现基底细胞在前列腺发育以及肿瘤发生过程中可以通过不对称分裂产生腔细胞,而腔细胞则只进行对称分裂从而维持和扩增腔细胞群。这一发现很好的解释了前列腺发育过程中的谱系发生问题,同时为鉴定前列腺癌细胞起源提供直观的细胞生物学证据。
[Abstract]:Background: stem cells divide symmetrically and asymmetrically in tissue development. Regeneration and tumorigenesis play an important role although the regulatory mechanisms of stem cell division patterns have been extensively studied in the neural tissues of the model animals Drosophila melanogaster and nematodes as well as mammals. However, in mammalian non-neural tissues or organs, the pattern of stem cell division is rarely reported. We systematically study the pattern of basal and lumen cell division during normal development of prostate and tumorigenesis by cell fate analysis and cell lineage tracing. Objective: to study the cell division pattern of basal cells and cavity cells during normal prostate development and tumorigenesis. To elucidate the polarity of basal cells and cavity cells during normal development and regeneration of prostate. Key issues such as cell spindle localization and cell fate determination. 2) study on the fate of basal cells and cavity cells by means of cell lineage tracing and basal cell knockout mice. To study whether Pten deletion affects the cell polarity of basal cells and cavity cells. Materials and methods: we use specific antibodies from a variety of prostatic epithelial basal cells and lumen cell markers. Antibodies to cell division markers and specific antibodies to cell polarity identified the polarity patterns and spindle localization patterns of basal cells and cavity cells. Lineages of basal and luminal cells, including prostatic cells, were traced to mice. Basal cell lineage knockout mice and prostate specific Pten knockout mice. We capture the fate of basal and cavity cells by positive fluorescence microscopy and laser confocal microscopy. Results: chapter 1. We observed that basal cells and cavity cells had different cell polarity patterns and spindle localization patterns. In chapter 2, the horizontal division of basal cells was proved to be symmetrical amplification division by cell fate analysis. The vertical division is asymmetric division, producing differentiated cavity cells. In contrast, the cavity cells only divide symmetrically. The progenitor cells produced are all cavity cells. Lineage tracing experiments further verify the bipotency of basal stem cells and the monpotency of cavity cells. In chapter 3, we prove that after knocking out Pten. The symmetrical division of the cavity cell itself can accelerate the expansion of the lumen cell group and make the prostate cancer present the cavity phenotype. The transformed basal cells can still undergo asymmetric division to produce cavity cells and induce cavities like prostate neoplasms. Chapter 4th. We found that the expression of Sox2 and Trop2 in the cavity cells increased during the development of prostate tumors, through the analysis of lineage analysis and cell division pattern analysis. It is proved that cavities can be generated by basal cells in lineage. Conclusion: the pattern of prostatic epithelial cell division is studied. We found that basal cells can produce cavities through asymmetric division during prostate development and tumorigenesis. While the cavity cells only divide symmetrically to maintain and expand the lumen cell population. This finding is a good explanation for the prostatic development of the lineage problems. It also provides direct cell biological evidence for identifying the origin of prostate cancer cells.
【学位授予单位】：上海交通大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R737.25

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