Modulation of Prostate Cancer Cell Motility by Chemopreventive Agent Genistein

金雀异黄素是一种NCI高优先级前列腺癌(PCA)化学预防药物。我们的预赛 研究表明，金雀异黄素抑制前列腺癌细胞脱离和侵袭，这是 转移性级联。我们假设金雀异黄素还可以通过抑制Pca的转移来抑制其转移。 前列腺癌细胞从前列腺癌进入人体血液循环。 在初步的体外研究中，我们证明了金雀异黄素抑制细胞前运动性的激活。 信号转导p38-HSP27(热休克蛋白27)途径，同时增强抗运动性ALK-2的激活 信号通路。在小鼠中，我们已经证明金雀异黄素抑制人前列腺癌细胞分离和 转移。我们的第一个孢子试验是一期临床试验，它定义了染料木素在前列腺癌中的药理作用 病人。我们的第二个孢子试验是前列腺摘除前的第二阶段设计。它证明了(1) 金雀异黄素耐受性良好，(2)它抑制了前列腺细胞的脱落，(3)它选择性地 调节前列腺细胞运动的基因。 在这项建议中，我们提出了三个目标： 1)确定金雀异黄素是否影响人类运动相关基因和调节蛋白 前列腺组织。使用我们第二次孢子试验中储存的前列腺组织，我们的研究将确定 金雀异黄素是否改变了前列腺细胞中相关调控通路的功能。 2)在动物模型中评估热休克蛋白27对金雀异黄素是否有影响 抑制转移的能力。经过改造表达HSP27水平改变的人前列腺癌细胞将是 通过原位植入小鼠体内。我们将评估他们形成转移的能力，无论有没有 染料木素治疗。 3)进行II期临床试验，以确定金雀异黄素是否抑制PCa细胞从 高危前列腺术前患者的前列腺进入循环。我们将实施第三个 孢子试验来验证这一假设。可测量的临床局限性高危PCa受试者 循环中的前列腺细胞将被累积到预期的第二阶段，染料木素与 安慰剂。金雀异黄素对前列腺细胞循环水平的影响将通过 PSA定量RT/PCR检测。

Genistein is a NCI high priority putative prostate cancer (PCa) chemopreventive agent. Our preliminary studies demonstrate that genistein inhibits PCa cell detachment and invasion, which are initial steps in the metastatic cascade. We hypothesize that genistein will also inhibit PCa metastasis by inhibiting the movement of prostate cancer cells from the prostate gland into the circulation in man. In preliminary in vitro studies, we demonstrated that genistein inhibits activation of the pro-cell-motility signaling p38-HSP27 (heat shock protein 27) pathway, while enhancing activation of the anti-motility ALK-2 signaling pathway. In mice, we have shown that genistein inhibits human PCa cell detachment and metastasis. Our first SPORE trial was a phase I clinical trial, and it defined genistein's pharmacology in PCa patients. Our second SPORE trial was a phase 2 pre-prostatectomy design. It demonstrated that (1) genistein was well tolerated, (2) that it inhibited prostate cell detachment, and (3) that it selectively modulated genes that regulate prostate cell motility. In this proposal, we propose three Aims: 1) To determine whether genistein affects motility associated genes and regulatory proteins in human prostate tissue. Using banked prostate tissue from our second SPORE trial, our studies will determine whether genistein alters the function of relevant regulatory pathways in prostate cells. 2) Evaluate in an animal model whether modulation of pro-cell-motility HSP27 has an impact on genistein's ability to inhibit metastases. Human PCa cells engineered to express altered levels of HSP27 will be orthotopically implanted into mice. We will evaluate their ability to form metastasis, with and without genistein treatment. 3) To conduct a Phase II clinical trial to determine whether genistein inhibits movement of PCa cells from the prostate gland into the circulation in high risk pre-prostatectomy patients. We will implement a third SPORE trial to test this hypothesis. Subjects with clinically-localized, high-risk PCa with measurable circulating prostate cells will be accrued on to a prospective phase 2, randomized trial of genistein versus placebo. The resultant effects of genistein on circulating levels of prostate cells will be assessed by a quantitative RT/PCR assay for PSA.