HORMONE REGULATED GENE IN OVARIAN CARCINOGENESIS

卵巢癌发生中的激素调控基因

• 批准号: 2895455
• 负责人: SAMUEL C MOK
• 金额: $ 18.69万
• 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 2001-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2895455
• 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.

流行病学数据支持这一假设，即卵巢癌可能是一种恶性肿瘤。 内分泌相关肿瘤然而，激素调节基因在 卵巢癌的发生还没有被广泛研究。，也称为 骨粘连蛋白，BM-40，43 K蛋白，是一种富含半胱氨酸的酸性成分 细胞外基质的细胞外基质， 序列保守性，并已被证明是直接调节， 孕激素和地塞米松以及细胞因子的间接作用。实验 在这项提案中概述的将测试的假设，这种孕酮 调节性糖蛋白β是卵巢表面的生理调节因子 上皮细胞的功能和卵巢上皮细胞的失调在卵巢癌中起作用。 致癌作用我们已经从正常卵巢中克隆了hepatocellular carcinoma基因， 上皮细胞，并证明它在高水平表达， 人正常卵巢表面上皮（HOSE）细胞和低得多的 在体外和体内卵巢癌细胞中的水平。 我们的数据显示 在正常人中， 在体外和体内的肿瘤细胞，并建议ESTA可能发挥作用， 在卵巢上皮细胞生长和分化中起重要作用。 基于这些初步数据，我们提出（I）。去研究 在正常卵巢和卵巢肿瘤组织中， 北方和Western blot检测不同分期和组织学分级 分析、原位杂交和免疫组化检测：（2）。到 定量正常软管分泌的生物活性物质的量 细胞扩散试验和北方 和蛋白质印迹分析：并表征 [~ 3 H]-胸苷掺入法测定细胞凋亡率， 膜侵入测定;和（3）。通过基因改变 正常HOSE细胞和卵巢癌细胞通过转染完整的 在表达载体中扩增反义和正义cDNA， 验证卵巢癌细胞中的hepatocellular carcinoma表达 改变恶性性质，如肿瘤生长和体外侵袭， in vivo . 如果将前列腺素确定为卵巢癌的重要调节因子， 卵巢上皮细胞功能及蛋白表达上调 癌细胞可以抑制其生长，这可能表明， 可能具有治疗潜力， 卵巢恶性肿瘤