CANCER STEM CELL LONGEVITY AND METASTATIC POTENTIAL IN BLADDER CANCER

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 最近对干细胞样癌细胞（SLCC）的研究提出了当前癌症疗法疗效低下的潜在原因以及改进方法。 目前的癌症化学疗法和放射疗法针对肿瘤中快速分裂的细胞，然而肿瘤是异质细胞群。 SLCC 包含少量细胞群，保持相对静止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，并且对化疗和放疗具有抵抗力。 SLCC 在治疗后仍然存在，并保留启动癌细胞增殖的能力；因此，人们相信 SLCC 是转移和复发的根源。 然而，对于 SLCC 的定义及其在癌症发生和进展中的作用存在争议。 使用膀胱癌作为模型，我们的目标是证明膀胱癌肿瘤中的 SLCC 比肿瘤主体更古老，对传统化疗和放疗具有抵抗力，并且是转移和复发的原因。 该项目的第一部分涉及从三种膀胱癌细胞系中分离出对化疗药物顺铂有一系列反应的干细胞群。 除了通过集落形成试验、长期增殖、分化以及对化疗和放疗的反应所证明的与干细胞相关的特性的能力之外，还将检查这些细胞的循环。 最初，我们计划分离 CD44、CD66C- 作为我们的干细胞群，因为这在文献中得到了最有力的支持。 据报道，侧向散射群体在几种膀胱癌系中具有干细胞能力，也可以进行研究。 该项目的第二个目标是研究新鲜人类膀胱肿瘤样本中的 SLCC。 从新鲜肿瘤样本中分离出的 SLCC 以及非干细胞的年龄将使用 AMS 的 14C 炸弹曲线来确定。这些分选细胞的 DNA 中 14C/C 的升高将用于指定合成日期。 可以将合成日期与患者的病史进行比较，以确定 SLCC 在肿瘤发展时间轴上的位置。 最后，我们将通过生产带有高度标记 DNA 的 SLCC 来开发一种用于癌症研究的新工具。 标记的癌症干细胞将根据干细胞的百分比和细胞周期特性，从项目初始部分使用的细胞系之一中产生。 这些标记的 SLCC 随后可用于研究其转移潜力，并作为癌症研究的示踪剂。 AMS 将使我们能够追踪单个细胞。 这项工作有能力改变当前用于治疗癌症的方法。 现有的疗法主要是针对构成肿瘤主体的高度增殖的细胞而开发的。 通过明确测量 SLCC 的寿命，可以确认它们是复发和转移的原因。 标记 SLCC 的创建将为癌症研究提供新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.