Exploring the role of progestin and vitamin D on inflammation and oxidative stress in in vitro and in vivo models of ovarian cancer

项目摘要/摘要 在卵巢癌发生的早期阶段，现在认为菌毛获得了分子 以及暴露于炎症介质和相关氧化应激时的遗传变化 (活性氧种-ROS)在排卵过程中从相邻卵巢释放。这些 变化导致“P53特征”--突变型P53蛋白积聚(最早的基因 卵巢癌的改变)，在海马伞的分泌细胞中最终导致 浆液性输卵管上皮内癌(STIC)的发生发展 脱落/剥落直接进入邻近的卵巢并进入腹膜。P53 在炎症、氧化和DNA中毒的过程中演变的特征和STIC损害 微环境具有明显的特征，包括极性丧失，上皮丛生， 细胞核多形性、P53异常表达和高增殖指数。发现了 这些损伤和它们如何发展打开了制定药理学策略的大门 这阻止和/或扭转了海流带早期的变革性事件，具有巨大的潜力 通过预防降低卵巢癌的发病率和死亡率。 以前，我们的工作主要集中在卵巢表面上皮细胞作为生物学靶点。 使用孕激素和维生素D预防卵巢癌，然而，关于 它们在FTE中的生物效应。这项研究的目的是探讨 孕激素和维生素D对卵巢癌早期致癌中炎症和氧化应激的影响 卵巢癌在体外和体内的疾病模型。我们假设抑制作用 炎症和氧化作用可能包括一种生物机制，有助于 这些制剂对癌症的预防作用。此外，我们将研究这一组合是否 黄体酮和维生素D的结合比任何一种单独使用都更有效。为了检验这一假设，我们 将利用来自父母资助的以下样本来评估炎症和氧化 孕激素和维生素D应激途径：(1)原代细胞培养 FTE以及含有显性负突变体P53的转化FTE，并 由绝经前和绝经后接受良性妇科手术的妇女建立 适应症，(2)输卵管石蜡切片和相应的分离株(RNA，DNA， 从小鼠(mogp-tag和Dird-PTEN)中获得的蛋白质和血浆/血清) 孕激素和维生素D的预防试验

Project Summary/Abstract In the early stages of ovarian carcinogenesis, it is now thought that the fimbria acquires molecular and genetic alterations when exposed to inflammatory mediators and associated oxidative stress (reactive oxygen species-ROS) released during ovulatory events from the adjacent ovary. These changes lead to the “p53 signature”- accumulation of mutant p53 protein (earliest genetic alteration in ovarian cancer), in the secretory cells of the fimbria ultimately leading to the development of serous tubal intraepithelial carcinoma (STIC), which spreads via shedding/exfoliation directly to the adjacent ovary and into the abdominal cavity. The p53 signatures and STIC lesions which evolve in an inflammatory, oxidative and DNA-toxic microenvironment are characterized by distinct features including loss of polarity, epithelial tufting, pleomorphic nuclei, abnormal p53 expression and a high-proliferative index. The discovery of these lesions and how they develop opens the door to the formulation of pharmacologic strategies that arrest and/or reverse the early transformative events in the fimbria, with immense potential to decrease ovarian cancer incidence and mortality through prevention. Previously, our work has focused primarily on the ovarian surface epithelium as the biologic target for ovarian cancer prevention using progestins and vitamin D. However, little is known regarding their biologic effects in the FTE. The objective of this study is to explore the impact of progestin and vitamin D on inflammation and oxidative stress in early carcinogenesis of ovarian cancer in in vitro and in vivo models of the disease. We hypothesize that inhibition of inflammatory and oxidative effects may comprise a biologic mechanism contributing to the cancer preventive effects of these agents. Furthermore, we will examine whether the combination of progestin and vitamin D is more effective than either agent alone. To test this hypothesis, we will utilize the following samples from the parent grant to evaluate inflammatory and oxidative stress pathways following administration of progestin and vitamin D: (1) cell cultures of primary FTE as well as transformed FTE harboring dominant negative mutant p53 obtained and established from pre- and postmenopausal women undergoing gynecologic surgery for benign indications, (2) paraffin sections of the fallopian tube and corresponding isolates (RNA, DNA, protein and plasma/serum) obtained from the mice (mogp-TAg and Dicer-PTEN) undergoing prevention trials with progestins and vitamin D.