DISSECTING COX-1 RELATED GENE PATHWAYS IN OVARIAN CANCER

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 摘要 卵巢癌是美国妇科癌症中死亡的主要原因。 由环氧合酶-1（考克斯-1）和环氧合酶-2（考克斯-2）介导的前列腺素（PG）途径与排卵有关，并已被认为是卵巢癌病因学中的潜在因素。 初步数据：我们已经利用cDNA微阵列比较正常卵巢组织样本与卵巢癌，通过这个过程已经显示考克斯-1的表达水平在癌症中高出2倍以上。 我们的合作者发表的数据同样表明，考克斯-1，而不是考克斯-2，在卵巢恶性肿瘤中高度表达。 假设和目的：本研究的长期目标是确定考克斯-1在卵巢癌发生中的具体作用。 我们假设在卵巢癌中，考克斯-1诱导PGE 2的产生，PGE 2靶向磷脂酰肌醇3激酶/蛋白激酶B（PI 3 K/Akt）信号传导以及参与肿瘤生长和进展的该途径的组分。 我们建议使用定制的组织阵列进行免疫组织化学（IHC）染色，以测量考克斯-1、考克斯-2和与PI 3 K相关的分子标记物：磷酸化Akt（pAkt）、缺氧诱导因子（HIF-1）和血管内皮生长因子（VEGF）的表达水平。 将进行平行的体外实验，通过用选择性考克斯-1抑制剂SC-560处理表达考克斯-1的细胞系OVCAR 3，进一步评价考克斯-1的功能。 我们将测量PGE 2代谢、细胞生长、凋亡、迁移和侵袭，以及PI 3 K、pAkt、HIF-1 <$和VEGF的表达水平。 我们还将利用cDNA微阵列来产生与考克斯-1相关的基因谱。 重要性：如果我们确定考克斯-1确实是卵巢癌肿瘤进展的一个突出因素，这项研究可能为将来将其用作治疗这种致命疾病的生物标志物和靶点提供基础。 A.具体目标。 由环氧合酶-1（考克斯-1）和环氧合酶-2（考克斯-2）介导的前列腺素（PG）途径与排卵有关，并已被认为是卵巢癌病因学中的潜在因素。 本研究的长期目标是确定考克斯-1在卵巢癌发生中的具体作用。 具体目标1： 分析卵巢组织中考克斯-1的表达与肿瘤发生的关系。 具体目标2： 探讨卵巢癌细胞中PGE 2和PI 3 K/Akt信号通路介导的细胞生长和血管生成对考克斯-1表达的影响。

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.