CANCER STEM CELL LONGEVITY AND METASTATIC POTENTIAL IN BLADDER CANCER

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 最近对干细胞样癌细胞(SLCC)的研究表明，目前癌症治疗效果低的一个潜在原因是一种改进方法。目前的癌症化疗和放射治疗针对的是肿瘤中快速分裂的细胞，然而肿瘤是一个异质的细胞群体。SLCC由一小部分人组成，保持相对静止的状态，并启动占肿瘤块大部分的增殖细胞。这些SLCC基本上处于休眠状态，对化疗和放射治疗具有抵抗力。SLCC在治疗后仍然存在，并保持其启动癌细胞增殖的能力；因此，人们认为SLCC是转移和复发的来源。然而，关于SLCC的定义及其在癌症发生发展中的作用仍存在争议。以膀胱癌为模型，我们的目标是显示膀胱癌中的SLCC比肿瘤的大部分年龄要大得多，对传统的化疗和放射治疗具有抵抗力，并与转移和复发有关。 该项目的第一部分涉及从对化疗药物顺铂有一系列反应的三种膀胱癌细胞系中分离干细胞群。除了通过克隆形成试验、长期增殖、分化以及对化疗和辐射的反应证明它们与干细胞相关的特性外，还将检查这些细胞的周期。最初，我们计划分离CD44+、CD66C-作为我们的干细胞种群，因为这在文献中得到了最强有力的支持。侧散射群也被报道在几个膀胱癌株中具有干细胞能力，也可能被研究。 该项目的第二个目标是在新鲜的人膀胱癌样本中研究SLCC。从新鲜肿瘤样本和非干细胞中分离出的SLCC的年龄将使用AMS的14C炸弹曲线来确定。这些分类细胞DNA中14C/C的升高将被用来指定合成日期。这一合成日期可以与患者的病史进行比较，以确定SLCC在肿瘤发展时间线上的位置。 最后，我们将通过生产带有高标记DNA的SLCC来开发一种用于癌症研究的新工具。根据干细胞的百分比和细胞周期特性，标记的癌症干细胞将从项目初始部分使用的一种品系中生产出来。这些标记的SLCC然后可以用于研究其转移潜力，并作为示踪剂用于癌症研究。AM将允许我们跟踪单个细胞。 这项工作有能力改变目前用于治疗癌症的方法。现有的治疗方法主要是针对构成肿瘤主体的高增殖细胞而开发的。通过明确地测量SLCC的生存期，它们可以被确认为复发和转移的原因。标记SLCC的创建将为癌症研究提供一个新的工具，并改变该领域的方向。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Recent studies on stem-like cancer cells (SLCC) suggest a potential reason for the low efficacy of current cancer therapies and a way to improve them. Current cancer chemo- and radio-therapies target the rapidly-dividing cells in a tumor, however a tumor is a heterogeneous population of cells. Comprising a small amount of the population, SLCC remain relatively static and initiate the proliferative cells that account for the majority of the tumor mass. These SLCC are essentially dormant and resistant to chemo- and radio-therapy. SLCC remain after therapy and retain their ability to initiate proliferative cancer cells; therefore it is believed SLCC are the source of metastasis and relapse. However, there is controversy over the definition of SLCC and their role in cancer development and progression. Using bladder cancer as a model, we aim to show SLCC in bladder cancer tumors are much older than the bulk of the tumor, resistant to conventional chemo- and radio-therapy and responsible for metastasis and relapse. The first part of the project involves isolating stem cell populations from three bladder cancer cell lines with a range of responses to the chemotherapeutic agent, cisplatin. The cycling of these cells will be examined in addition to their ability to demonstrate properties associated with stem cells as demonstrated through colony-forming assays, long term proliferation, differentiation and response to chemotherapy and radiation. Initially we plan to isolate CD44+,CD66C- as our stem cell population, as this has the strongest support in the literature. A side-scatter population has also been reported to possess stem cell capabilities in several bladder cancer lines and may also be investigated. The second goal of the project is to investigated SLCC in fresh human bladder tumor samples. The age of SLCC isolated from fresh tumor samples along with non-stem cells will be determined using the 14C bomb curve with AMS. The elevation of 14C/C in the DNA of these sorted cells will be used to assign a date of synthesis. This date of synthesis can be compared to the medical history of the patient in order to determine where on the timeline of tumor development SLCC fall. Finally, we will develop a new tool for use in cancer research by producing SLCC with highly labeled DNA. Labeled cancer stem cells will be produced from one of the lines used in the initial part of the project based upon the percentage of stem cells and cell cycle properties. These labeled SLCC can then be used in research on their metastatic potential and as a tracer in cancer research. AMS will allow us to follow individual cells. This work has the power to transform the current methods used to treat cancer. Existing therapies are developed primarily against the highly proliferative cells that make up the bulk of the tumor. By definitively measuring the longevity of SLCC, they can be confirmed as the reason for relapse and metastasis. The creation of labeled SLCC will provide a new tool to be used in cancer research and change the direction of the field.