CANCER STEM CELL LONGEVITY AND METASTATIC POTENTIAL IN BLADDER CANCER

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH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 中心，不一定是研究者的机构。 最近对干细胞样癌细胞（SLCC）的研究表明了当前癌症治疗疗效低下的潜在原因以及改善它们的方法。 目前的癌症化疗和放疗靶向肿瘤中快速分裂的细胞，然而肿瘤是异质细胞群。SLCC包含少量的群体，保持相对静止并启动占肿瘤块大部分的增殖细胞。 这些SLCC基本上是休眠的，对化疗和放疗有抵抗力。 SLCC在治疗后仍然存在，并保留其启动增殖癌细胞的能力;因此，认为SLCC是转移和复发的来源。 然而，对于SLCC的定义及其在癌症发生和发展中的作用存在争议。 使用膀胱癌作为模型，我们的目标是显示膀胱癌肿瘤中的SLCC比肿瘤的大部分年龄大得多，对传统的化疗和放疗具有抗性，并负责转移和复发。 该项目的第一部分涉及从三个膀胱癌细胞系中分离干细胞群，这些细胞系对化疗药物顺铂有一系列反应。 除了通过集落形成试验、长期增殖、分化和对化疗和放疗的反应证明与干细胞相关的特性之外，还将检查这些细胞的循环。 最初，我们计划分离CD 44+，CD 66 C-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。 也有报道称，侧向散射群体在几种膀胱癌细胞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膀胱肿瘤样品中的SLCC。 从新鲜肿瘤样品中分离的SLCC与非干细胞沿着的年龄将使用AMS的14 C炸弹曲线来确定。这些分选细胞DNA中14 C/C的升高将用于指定合成日期。 可以将该合成日期与患者的病史进行比较，以确定SLCC在肿瘤发展的时间轴上的位置。 最后，我们将开发一种用于癌症研究的新工具，通过生产高度标记DNA的SLCC。 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一个细胞系中产生。 这些标记的SLCC可以用于研究其转移潜力，并作为癌症研究中的示踪剂。AMS将使我们能够跟踪单个细胞。 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针对构成肿瘤大部分的高度增殖细胞。 通过明确测量SLCC的寿命，可以证实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一种新的工具，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.