COLLAGEN TYPE I/SPHINGOLIPID PATHWAYS IN REGULATION OF LEIOMYOMA GROWTH

I 型胶原蛋白/鞘脂途径调节平滑肌瘤生长

• 批准号: 6206497
• 负责人: SHELIA A. MCCLURE
• 金额: $ 20.17万
• 依托单位: SPELMAN COLLEGE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2000-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6206497
• 关键词: collagen; enzyme linked immunosorbent assay; estradiol; growth factor; high performance liquid chromatography; hormone regulation /control mechanism; human tissue; leiomyoma; neoplastic growth; sphingolipids; thin layer chromatography; uterus preneoplastic state

Uterine leiomyomas, also known as fibroid tumors, are the most common tumors of the uterus, occurring with a frequency of 20-25% in women of reproductive age. Their occurrence has been linked to diverse symptoms including excessive uterine bleeding, pelvic pressure and pain, premature births, and infertility. The increased incidence of leiomyomas after the onset of menarche, the growth of tumors during pregnancy and after administration of steroids as well as the regression of tumors after menopause gives clinical evidence to support the role of ovarian steroids in the growth of uterine fibroids. These tumors also exhibit growth- factor dependent growth potential. Three human uterine cell lines designated ST1 MT1 and HT1 have been established in our laboratory. The long-term objectives of this research are to identify the factors involved in the regulation of the proliferation of uterine leiomyomas utilizing these cell lines and to determine the mechanism of action of these factors. The specific aims of the proposed research are to: I. Determine if the presence of collagen type I in the extracellular matrix of leiomyomas has an effect on the response of these tumors to known mitogens such as estradiol, EGF and mitogenic factors from conditioned medium from ST1 cells. In these studies ST1 and HT1 cells will be grown on defined matrices. The level of collagen type I secretion by ST1 and HT1 cells grown on these matrices will be determined by ELISA. The effect of estradiol, EGF and the mitogenic fraction of conditioned medium on the growth kinetics of these cell lines will also be assessed, when cells are grown on defined matrices. II. Purify the low molecular weight mitogenic factor from conditioned medium of ST1 cell lines and determine if the factor is a member of the sphingolipid class of growth factors. The content of sphingosine 1- phosphate in the mitogenic fraction of conditioned medium will be determined by HPLC to determine any correlation between this activity and sphingosine 1-phosphate will also be assessed. III. Determine if growth factors for uterine leiomyomas act via a sphingolipid signaling pathways in the regulation of leiomyoma growth. In the proposed studies, ST1 and HT1 cell lines will be exposed to mitogens. Levels of all intermediates in the putative signal transduction pathway (ceramide, sphingosine and sphingosine 1-phosphate) will be monitored by thin-layer chromatography and/or HPLC.

子宫肌瘤，也被称为肌瘤，是最常见的 子宫肿瘤，发生频率为20%-25% 生育年龄。它们的发生与不同的症状有关 包括子宫过度出血、盆腔压迫和疼痛、早产 出生和不孕不育。术后肌瘤发病率的增加 月经初潮的开始，妊娠期间及以后肿瘤的生长 激素的应用以及术后肿瘤的消退 更年期提供临床证据支持卵巢类固醇的作用 子宫肌瘤的生长。这些肿瘤也表现出生长- 依赖于因素的增长潜力。三种人子宫细胞系 命名为ST1、MT1和HT1的基因已在本实验室建立。这个 这项研究的长期目标是确定所涉及的因素 在子宫肌瘤增殖调控中的作用 并确定这些细胞系的作用机制 各种因素。拟议研究的具体目标是：一、确定 如果I型胶原在细胞外基质中的存在 肌瘤对这些肿瘤对已知有丝分裂原的反应有影响 如雌二醇、表皮生长因子和有丝分裂因子 ST1细胞。在这些研究中，ST1和HT1细胞将在定义的 矩阵。ST1和HT1细胞分泌I型胶原的水平 在这些基质上生长的将通过酶联免疫吸附试验进行检测。的影响 雌二醇、EGF和条件培养液中促有丝分裂成分对细胞生长的影响 还将评估这些细胞系的生长动力学，当细胞 在定义的矩阵上生长。二、纯化低分子量促有丝分裂原 从ST1细胞系的条件培养液中提取因子，并确定 因子是鞘磷脂类生长因子中的一员。这个 黄连有丝分裂组分中1-磷酸鞘氨醇的含量 条件培养液将通过高效液相色谱法进行测定，以确定是否存在相关性 这一活性和1-磷酸鞘氨醇之间的关系也将得到评估。 确定子宫肌瘤的生长因子是否通过 鞘磷脂信号通路在调节肌瘤生长中的作用。在……里面 在拟议的研究中，ST1和HT1细胞株将暴露于有丝分裂原。 可能的信号转导途径中所有中间体的水平 (神经酰胺、鞘氨醇和1-磷酸鞘氨醇)将由 薄层层析法和/或高效液相色谱法。