MARKERS OF BLADDER CANCER AND THEIR MODULATION BY DFMO

作为最近资助的一份合同(N01 CN-25434-01)的配套合同 浅表性移行细胞癌的化学预防 膀胱，我们建议研究TCC的中间分子标记。 以及不可逆的二氟甲基鸟氨酸(DFMO)对它们的调节 多胺(PA)合成酶抑制剂鸟氨酸 脱羧酶(ODC)。在N01 CN-25434中，40例移行细胞癌和20例非移行细胞癌患者 将测定2剂DFMO(或安慰剂)的生物效应 在相关靶点(恶性和正常尿路上皮细胞和组织， 和尿液)，以确定生物活性DFMO剂量用于 安慰剂对照，第三阶段，240例患者的预防性试验 完全切除浅表移行细胞癌。目前的调查将 利用这些研究中的主题，专注于相互作用的分子 已知在TCC和/或ODC阻滞剂/PA中改变的通路 耗尽。这些包括：1)PA代谢；2)相互作用 尿表皮生长因子及其尿路上皮受体 已知可诱导TCC细胞中的ODC活性；以及3)蛋白激酶 C(PKC)是EGF信号和ODC的重要组成部分 激活。 尿路上皮组织和尿液中乙酰化和未结合PA的水平 (由高效液相色谱法测定)将与尿路上皮ODC活性相关 (放射生物测定)表达(免疫组织学)在有无症状患者中的表达 TCC和那些正在接受和没有接受DFMO的人。尿液[EGF](RIA) 和尿路上皮EGF-R的表达(免疫组织学/免疫细胞学) 患者将与影响(例如尿路)的因素相关 PH)，或由EGF/EGF-R的增殖(如Ki67染色)引起的 相互作用，以及与PA/ODC参数。这些也将是相关的 具有主要的PKC亚型的组织图谱(免疫组织学) 已知在正常细胞和恶性肿瘤细胞中有差异表达 在非尿路上皮组织中。 通过比较患有和不患有TCC的患者的这些参数，以及 特别是通过以纵向方式重复测量，其中 正在监测疾病结局，疾病及其影响 复发以及ODC封锁将得到更好的定义。这些 研究应提供有关分子的重要临床信息 恶性尿路上皮转化、PA代谢和DFMO的标记物 TCC的生物学效应以及对生物学的见解 发展、成长和复发。

As a companion to a recently funded contract (N01 CN-25434-01) for the chemoprevention of superficial transitional cell cancer (TCC) of the bladder, we propose to investigate intermediate molecular markers of TCC and their modulation by difluoro-methylornithine (DFMO), an irreversible inhibitor of the polyamine (PA) synthesizing enzyme, ornithine decarboxylase (ODC). In N01 CN-25434, 40 TCC and 20 non-TCC patients will have the biologic effects of 2 doses of DFMO (or placebo) determined in relevant targets (malignant and normal urothelial cells and tissues, and urine) to determine a biologically active DFMO dose to use in a placebo-controlled, Phase III, preventative trial in 240 patients with completely resected superficial TCC. The present investigation will utilize subjects in these studies to focus on interacting molecular pathways which are known to be altered in TCC and/or with ODC blockade/PA depletion. These include: 1) Pa metabolism; 2) the interaction of urinary epidermal growth factor (EGF) and its urothelial receptor (EGF-R) which is known to induce ODC activity in TCC cells; and 3) protein kinase C (PKC) which is an integral component of EGF signaling and ODC activation. Acetylated and unconjugated PA levels in urothelial tissues and urine (determined by HPLC) will be correlated with urothelial ODC activity (radiobioassay) expression (immunohistology) in patients with and without TCC and in those who are and are not receiving DFMO. Urinary [EGF] (RIA) and urothelial EGF-R expression (immunohistology/immunocytology) in these patients will be correlated with factors that influence (e.g. urinary pH), or result from (e.g. proliferation [Ki67 staining]) the EGF/EGF-R interaction, and with PA/ODC parameters. These will also be correlated with tissue profiles of predominant PKC isoforms (immunohistology) which are known to be differentially expressed in normal versus malignant cells in non-urothelial tissues. By comparing these parameters in patients with and without TCC, and particularly by repeating measurements in a longitudinal fashion in which disease outcome is being monitored, the effects of disease and its recurrence, as well as of ODC blockade will be better defined. These studies should provide important clinical information about molecular markers of malignant urothelial transformation, PA metabolism, and DFMO's biological effects as well as insights into the biology of TCC development, growth, and recurrence.