HORMONE REGULATED GENE IN OVARIAN CARCINOGENESIS

卵巢癌发生中的激素调控基因

• 批准号: 2700655
• 负责人: SAMUEL C MOK
• 金额: $ 18.15万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-05-01 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2700655
• 关键词: basement membrane; carcinogenesis; cell growth regulation; cell proliferation; clinical research; extracellular matrix; female; gene expression; hormone regulation /control mechanism; human subject; immunocytochemistry; in situ hybridization; laboratory mouse; neoplastic cell; northern blottings; osteonectin; ovary neoplasms; progesterone; protein structure function; tissue /cell culture; transfection; western blottings

Epidemiological data support the hypothesis that ovarian cancers may be an endocrine-related tumor. However, the role hormone regulated genes in ovarian carcinogenesis has not been extensively studied. SPARC, also termed osteonectin, BM-40, and 43K protein, is an acidic, cysteine-rich component of the extracellular matrix that displays a high degree of interspecies sequence conservation and has been shown to be directly regulated by progesterone and dexamethasone and indirectly by cytokines. The experiments outlined in this proposal will test the hypothesis that this progesterone regulated glycoprotein SPARC is a physiologic regulator of ovarian surface epithelial cell function and deregulation of SPARC plays a role in ovarian carcinogenesis. We have cloned the SPARC gene from the normal ovarian epithelial cells and demonstrated that it is expressed at high levels in the human normal ovarian surface epithelial (HOSE) cells and at much lower levels in ovarian carcinoma cells in vitro and in vivo. Our data show that there is a tight correlation between SPARC expression in normal and neoplastic cell in vitro and in vivo and suggest that SPARC may play an important role in ovarian epithelial cell growth and differentiation. Based on these preliminary data, we propose (I). To study the expression pattern of the SPARC gene in normal ovary and ovarian tumor tissues of different stages and histological grades by Northern and Western blot analysis, in-situ hybridization and immunohistochemical detection: (2). To quantify the amount of biological active SPARC secreted by normal HOSE cells and the ovarian carcinoma cells by cell spreading assay and Northern and Western Blot analysis: and to characterize the biological effects of SPARC on these cells by [3H]-thymidine incorporation study and basement membrane invasion assay; and (3). To genetically alter SPARC expression in normal HOSE cells and ovarian carcinoma cells by transfection of full length anti-sense and sense SPARC cDNA in expression vector in order to test the hypothesis that expression of SPARC in ovarian carcinoma cells alters malignant properties such as tumor growth and invasion in vitro and in vivo . If SPARC is identified as an important regulator of ovarian epithelial cell function and up-regulation of the protein in ovarian carcinoma cells can inhibit their growth, this may suggest that strategies based on alteration in SPARC expression may have therapeutic potential in ovarian malignancies.

流行病学数据支持以下假设：卵巢癌可能是一种 内分泌相关肿瘤。然而，激素调节基因的作用 卵巢癌的发生机制尚未得到广泛研究。 SPARC，也称为 骨连接蛋白、BM-40 和 43K 蛋白是一种富含半胱氨酸的酸性成分 显示出高度种间性的细胞外基质 序列保护并已被证明直接受 黄体酮和地塞米松，间接通过细胞因子。实验 该提案中概述的内容将检验以下假设：黄体酮 调节糖蛋白 SPARC 是卵巢表面的生理调节因子 上皮细胞功能和 SPARC 失调在卵巢中发挥作用 致癌作用。我们从正常卵巢中克隆了SPARC基因 并证明它在上皮细胞中高水平表达 人类正常卵巢表面上皮 (HOSE) 细胞，并且含量要低得多 卵巢癌细胞的体外和体内水平。 我们的数据表明 正常和正常状态下的 SPARC 表达之间存在紧密相关性 肿瘤细胞的体外和体内实验表明 SPARC 可能发挥着重要作用 在卵巢上皮细胞的生长和分化中起重要作用。 根据这些初步数据，我们提出（I）。研究表达 正常卵巢和卵巢肿瘤组织中 SPARC 基因的模式 通过 Northern 和 Western blot 检测不同阶段和组织学等级 分析、原位杂交和免疫组化检测：(2)．到 定量正常 HOSE 分泌的生物活性 SPARC 的量 细胞和卵巢癌细胞通过细胞扩散试验和Northern 和蛋白质印迹分析：并表征其生物效应 通过 [3H]-胸苷掺入研究和基础对这些细胞进行 SPARC 膜侵袭试验；和（3）。基因改变 SPARC 表达 全转染正常HOSE细胞和卵巢癌细胞 长度反义和正义 SPARC cDNA 在表达载体中，以便 检验卵巢癌细胞中 SPARC 表达的假设 改变恶性特性，例如体外肿瘤生长和侵袭， 体内。 如果 SPARC 被确定为卵巢的重要调节因子 卵巢上皮细胞功能和蛋白质上调 癌细胞可以抑制其生长，这可能表明策略 基于 SPARC 表达的改变可能具有治疗潜力 卵巢恶性肿瘤。