HORMONE REGULATED GENE IN OVARIAN CARCINOGENESIS

流行病学数据支持卵巢癌可能是一种 内分泌相关肿瘤。然而，荷尔蒙调节的基因在 卵巢癌的发生还没有得到广泛的研究。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 显示高度种间差异的细胞外基质 序列保守性，并已被证明直接受 黄体酮和地塞米松，并间接通过细胞因子。这些实验 这项提案中概述的内容将检验这样一个假设，即 调节糖蛋白SPARC是卵巢表面的一种生理调节因子 卵巢上皮细胞功能和SPARC基因失控在卵巢中的作用 致癌。我们从正常卵巢组织中克隆了SPARC基因 上皮细胞，并证明它高水平表达在 人正常卵巢表面上皮(Hose)细胞，并在更低的 卵巢癌细胞在体内外的表达水平。我们的数据显示 SPARC在正常组织和正常组织中的表达密切相关 并提示SPARC可能在肿瘤细胞中发挥重要作用。 在卵巢上皮细胞的生长和分化中起重要作用。 基于这些初步数据，我们建议(I)。要研究表达方式 卵巢癌患者正常卵巢和卵巢肿瘤组织中SPARC基因的图谱 不同分期、不同组织学分级的Northern和Western印迹分析 分析、原位杂交和免疫组织化学检测：(2)。至 测定正常软管中生物活性SPARC的分泌量 细胞铺展实验和Northern杂交检测卵巢癌细胞 和Western Blot分析：并表征其生物学效应 [~3H]-胸腺嘧啶核苷掺入法和底物对这些细胞的SPARC研究 (3)膜侵袭试验。在基因上改变SPARC的表达 Full基因转染正常软管细胞和卵巢癌细胞的研究 将反义和正义SPARC基因分别克隆到表达载体中 验证SPARC在卵巢癌细胞中表达的假说 在体外改变肿瘤生长和侵袭等恶性特性 在活体内。如果SPARC被认为是卵巢的重要调节因子 卵巢上皮细胞功能及其蛋白表达上调 癌细胞可以抑制其生长，这可能提示采取的策略 基于SPARC表达的改变可能具有治疗潜力 卵巢恶性肿瘤。

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.