Anoikis Effect by Quinazolines on Prostate Growth

描述(申请人提供)：在前列腺生长障碍中失去对细胞凋亡的控制涉及到细胞内凋亡信号通路下游效应物的表达/激活的改变。通过靶向这一途径中的特定成分来恢复功能性的凋亡机制，在调节前列腺生长方面具有重要意义。最近的研究证明，基于喹唑啉的A1受体拮抗剂多沙唑嗪和特拉唑嗪能够通过A1受体非依赖性机制诱导人前列腺平滑肌细胞、良性和恶性上皮细胞凋亡，并抑制组织血管。我们的假设是，基于喹唑啉的A1肾上腺素受体拮抗剂通过诱导细胞内信号效应分子诱导细胞凋亡来抑制前列腺生长，并通过促进失巢凋亡(通过干扰细胞外基质附着)抑制血管生成。为了验证这一假说，我们提出了以下目标：在特定的目标1中，我们将使用DNA微阵列分析和蛋白质组学分析来确定以喹唑啉为基础的A1受体拮抗剂诱导前列腺良恶性细胞凋亡的细胞内信号效应分子。具体目标2将确定参与喹唑啉诱导的前列腺上皮和内皮细胞失巢凋亡的细胞外基质成分，重点是Fas-Fasigand-caspase-8激活机制和整合素效应。具体目标3将确定在A1阻断治疗后，喹唑啉、多沙唑嗪和特拉唑嗪抑制患者的前列腺血管形成和组织血管生成。对接受A1阻滞剂治疗(针对梗阻症状)的BPH患者的组织样本进行免疫组织化学分析，使用抗第VIII因子(微交叉密度)、血管内皮生长因子(VEGF)和焦点粘附性激酶(FAK)的抗体。这项研究的目的是确定细胞内(转化生长因子-β)和细胞外基质(整合素)环境中的早期反应基因，这些基因是喹唑啉介导的细胞凋亡的基础，该药物干预对良、恶性前列腺生长具有靶向性。潜在的研究结果将证实，喹唑啉类药物的细胞凋亡/失巢凋亡效应在良性前列腺增生症(BPH)前列腺癌的发生和发展中具有重要的临床意义。

DESCRIPTION (provided by applicant): Loss of apoptotic control in prostate growth disorders involves the altered expression/activation of downstream intracellular effectors of apoptosis signaling pathways. Re-instating functional apoptotic mechanisms, by targeting specific components in this pathway, are of high significance in the regulation of prostate growth. Recent studies documented the ability for quinazoline-based a1-adrenoceptor antagonists, doxazosin and terazosin, to induce apoptosis in human prostate smooth muscle, benign and malignant epithelial cells, via an a1-adrenoceptor independent mechanism and suppress tissue vascularity. Our hypothesis is that quinazoline-based a1-adrenoceptor antagonists suppress prostate growth by inducing apoptosis (via induction of intracellular signaling effectors) and inhibit angiogenesis by promoting anoikis (via interference with extracellular matrix attachments). To test this hypothesis, the following aims are proposed: In Specific Aim 1 we will use DNA microarray analysis and proteomic analysis to identify intracellular signaling effectors of apoptosis induced by quinazoline-based a1-adrenoceptor antagonists in prostate benign and malignant cells. Specific Aim 2 will identify the extracellular matrix components involved in quinazoline-induced anoikis in prostate epithelial and endothelial cells with focus on Fas-Fasligand-caspase-8 activation mechanism and integrin effectors. Specific Aim 3 will establish that the quinazolines, doxazosin and terazosin, suppress prostate vasculature and inhibit tissue angiogenesis in patients after a1 blockade treatment. Immunohistochemical analysis will be performed on tissue specimens from BPH patients treated with a1 blockade (for obstructive symptoms) using antibodies against Factor VIII (microcrossed density), vascular endothelial growth factor (VEGF), and focal ashesion kinase (FAK). The goal of the proposed studies is to determine the early response genes at the intracellular (TGF-beta) and extracellular matrix (integrins) environment underlying the quinazoline-mediated apoptosis, targeted by this pharmacologic intervention in benign and malignant prostate growth. Potential results will establish the apoptotic/anoikis effect by the quinazolines as a significant phenomenon with clinical relevance in development and progression of benign prostatic hyperplasia (BPH) prostate cancer.