FORMS OF PROSTATE SPECIFIC ANTIGEN AND HK2 IN CANCER

我们寻求扩展有希望的研究，表明前列腺特异性 抗原(PSA)和腺激肽释放酶(HK2)以多种形式存在于 前列腺癌(CAP)患者的血液和血液指标的测定 PSA以PSAAlpha2-巨球蛋白形式存在的比例(Alpha2M) 提高早期前列腺帽与良性前列腺的鉴别能力 前列腺增生症(BPH)。这一点将得到充分表征的证实 预期的患者样本。此外，我们还将测量比例 在这些患者中检测其各种形式的PSA，并识别和测量 相关的新型帽标的格式，HK2。我们发现HK2存在 在帽状等离子体中以几种形式存在，包括HK2-α2M。临床化验 不要检测PSA-Alpha2M，也没有HK2的临床检测方法。我们 将确定哪种测量组合提供的最大 CAP与BPH的区别。由于PSA的形式在 早期与晚期CAP相比，我们的检测也可能对预后有帮助。这个 目标是提供简单的ELISA法，以减少侵入性， 诊断和ELISA所需的昂贵程序 预后，帮助医生选择治疗CAP的积极程度 患者，或确定BPH患者谁将在以后发生CAP。它 假设PSA和HK2以酶原的形式分泌，被激活 在需要的时候，然后用蛋白水解酶抑制剂灭活。这个 合成PSA和HK2的细胞的病理生理学在帽上不同 VS BPH，并可能在不同的CAP过程中。PSA之间的时间间隔 或HK2向血液的分泌和转位可能不同，并且 他们在前列腺中接触的酶和抑制剂是不同的 从鲜血中分离出来。因此，PSA和HK2的不同谱型 与良性前列腺增生症相比，会在CAP中发现，并在CAP的不同病程中发现。 为达致我们的目标，我们会：1.在CAP和BPH中确定HK2的形式 血浆，并开发相关HK2表格的检测。净化HK2和 制备相关HK2的抗体和标准HK2-抑制物 表格。2.前瞻性地采集300个帽子的系列血样 患者并记录完整的病历。从以下地点收集预期样本 300例PSA升高的未确诊患者。3.测量PSA-Alpha2M 每个样本的PSA-ACT、游离PSA和总PSA，并计算 每种PSA表格的比例。同样地，量度有关的HK2表格。 4.将各种形式的PSA和HK2的比例与 判断所有CAP患者病程对预后的价值 每一项测量的结果。5.将不同形式的 PSA和HK2对CAP或BPH患者继发诊断的意义 他们没有被诊断出来。确定每种方法的特异性和敏感性 测量以确定其诊断价值。技术将包括 蛋白质纯化、抗体及酶联免疫吸附试验、免疫印迹 酶活性测定和多肽合成。

We seek to extend promising studies showing that prostate specific antigen (PSA) and glandular kallikrein (hK2) occur in several forms in blood of prostate cancer (CaP) patients and that measurement of the proportion of PSA in the form of PSAalpha2-macroglobulin (alpha2M) improves the discrimination between early CaP and benign prostatic hyperplasia (BPH). This will be confirmed with well-characterized prospective patient samples. Moreover, we will measure the proportion of PSA in its various forms in these patients, and identify and measure forms of the related novel CaP marker, hK2. We found that hK2 exists in several forms in CaP plasmas, including hK2-alpha2M. Clinical assays do not detect PSA-alpha2M and there are no clinical assays for hK2. We will determine which combination of measurements provides the greatest discrimination between CaP and BPH. Since forms of PSA also vary in early vs. advanced CaP, our assays may also be useful in prognosis. The goal is to provide simple ELISA assays that will reduce the invasive, expensive procedures that are needed for diagnosis and ELISAs that are prognostic, to help physicians choose how aggressively to treat CaP patients, or to identify BPH patients who will develop CaP later. It is hypothesized that PSA and hK2 are secreted as zymogens, activated when needed, and then inactivated by protease inhibitors. The pathophysiology of cells that synthesize PSA and hK2 vary in CaP vs. BPH and possibly in different courses of CaP. The time between PSA or hK2 secretion and translocation to the blood may differ, and the enzymes and inhibitors to which they are exposed in the prostate differs from that in blood. Thus, a different spectrum of PSA and hK2 forms will be found in CaP compared to BPH, and in different courses of CaP. To meet our goals, we will: 1. Identify forms of hK2 in CaP and BPH plasmas and develop assays for relevant hK2 forms. Purify hK2 and prepare antibodies and standard hK2-inhibitor complexes for relevant hK2 forms. 2. Prospectively collect serial blood samples from 300 CaP patients and document full histories. Collect prospective samples from 300 undiagnosed patients with elevated PSA. 3. Measure PSA-alpha2M PSA-ACT, free PSA and "total" PSA in each sample and calculate the proportions of each PSA form. Similarly measure relevant hK2 forms. 4. Correlate the proportions of various forms of PSA and hK2 to the course of disease in all CaP patients to determine the prognostic value of each measurement. 5. Correlate the proportions of various forms of PSA and hK2 to the subsequent diagnosis of CaP or BPH in the patients who were undiagnosed. Determine the specificity and sensitivity of each measurement to determine its diagnostic value. Techniques will include protein purification, antibody and ELISA development, immunoblotting, enzyme activity assays, and peptide synthesis.