DISSECTING COX-1 RELATED GENE PATHWAYS IN OVARIAN CANCER

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 摘要 卵巢癌是美国妇科癌症的主要死因。由环氧合酶-1(COX-1)和环氧合酶-2(COX-2)酶介导的前列腺素(PG)途径与排卵有关，并被认为是卵巢癌发病的潜在因素。初步数据：我们利用基因芯片对正常卵巢组织样本和卵巢癌样本进行了比较，通过这一过程显示COX-1在癌症中的表达水平高出2倍以上。我们的合作者发表的数据同样表明，在卵巢恶性肿瘤中高表达的是COX-1，而不是COX-2。假设和目的：本研究的长期目标是确定COX-1在卵巢癌发生中的具体作用。我们假设，在卵巢癌中，COX-1诱导PGE2的产生，PGE2的靶向是磷脂酰肌醇3激酶/蛋白激酶B(PI3K/Akt)信号和该途径中参与肿瘤生长和进展的组件。我们建议使用定制的组织芯片免疫组织化学(IHC)染色来检测COX-1、COX-2的表达水平以及与PI3K相关的分子标志物：磷酸化Akt(PACT)、缺氧诱导因子(HIF-1)和血管内皮生长因子(VEGF)。通过选择性COX-1抑制剂SC-560处理表达COX-1的细胞系OVCAR3，将进行平行的体外实验，以进一步评估COX-1的功能。我们将检测PGE2代谢、细胞生长、凋亡、迁移和侵袭，以及PI3K、PAKT、HIF-1β和VEGF的表达水平。我们还将利用基因芯片来生成与COX-1相关的基因图谱。意义：如果我们确定COX-1确实是卵巢癌肿瘤进展中的一个重要因素，这项研究可能为未来将其用作生物标记物和治疗这一致命疾病的靶点提供基础。 A.具体目标。 由环氧合酶-1(COX-1)和环氧合酶-2(COX-2)酶介导的前列腺素(PG)途径与排卵有关，并被认为是卵巢癌发病的潜在因素。这项研究的长期目标是确定COX-1在卵巢癌发生中的具体作用。 目的1：探讨卵巢组织中环氧合酶-1的表达与肿瘤发生的关系。 特异性目的2：探讨PGE2和PI3K/Akt信号通路介导的COX-1表达对卵巢癌细胞生长和血管生成的影响。

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. ABSTRACT Ovarian cancer is the leading cause of death among gynecologic cancers in the United States. The prostaglandin (PG) pathway mediated by cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) enzymes is implicated in ovulation and has been suggested as a potential factor in the etiology of ovarian cancer. Preliminary Data: We have utilized cDNA microarrays to compare normal ovarian tissue samples to ovarian carcinomas, and through this process have shown expression levels of COX-1 to be more than 2-fold higher in the cancers. Data published by our collaborators similarly demonstrates that COX-1, not COX-2, is highly expressed in ovarian malignancies. Hypothesis and Aims: The long-range goal of this research is to determine the specific contributions of COX-1 to ovarian carcinogenesis. We hypothesize that in ovarian cancers, COX-1 induces PGE2 production, which targets phosphatidylinositol 3 kinase/protein kinase B (PI3K/Akt) signaling and components of this pathway that are involved in tumor growth and progression. We propose the use of custom-made tissue arrays for immunohistochemistry (IHC) staining to measure expression levels of COX-1, COX-2 and molecular markers associated with PI3K: phosphorylated Akt (pAkt), hypoxia induced factor (HIF-1¿) and vascular endothelial growth factor (VEGF). Parallel in vitro experiments will be performed to further evaluate COX-1 function by treating a COX-1 expressing cell line, OVCAR3, with a selective COX-1 inhibitor, SC-560. We will measure PGE2 metabolism, cell growth, apoptosis, migration and invasion, as well as expression levels of PI3K, pAkt, HIF-1¿ and VEGF. We will also utilize cDNA microarrays to generate gene profiles that are associated with COX-1. Significance: If we determine that COX-1 is indeed a prominent factor in tumor progression in ovarian cancer, this research may provide the basis for its future use as a biomarker and target for therapy in the management of this deadly disease. A. Specific Aims. The prostaglandin (PG) pathway mediated by cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) enzymes is implicated in ovulation and has been suggested as a potential factor in the etiology of ovarian cancer. The long-range goal of this research is to determine the specific contributions of COX-1 to ovarian carcinogenesis. SPECIFIC AIM 1: To analyze COX-1 expression and tumorigenesis in ovarian tissues. SPECIFIC AIM 2: To evaluate the effects of COX-1 expression on cell growth and angiogenesis, which are factors mediated by PGE2 production and PI3K/Akt signaling in ovarian cancer cells.