MESENCHYMAL DERIVED GROWTH FACTORS IN PROSTATIC CANCER

前列腺癌中的间充质衍生生长因子

• 批准号: 3202378
• 负责人: DAVID R ROWLEY
• 金额: $ 14.16万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-30 至 1997-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3202378
• 关键词: androgens; antibody formation; athymic mouse; cell growth regulation; clone cells; complementary DNA; enzyme linked immunosorbent assay; epithelium; gene deletion mutation; gene expression; genetic mapping; hormone related neoplasm /cancer; human tissue; immunocytochemistry; in situ hybridization; mesenchyme; molecular cloning; neoplasm /cancer genetics; neoplastic cell; northern blottings; nucleic acid probes; oligonucleotides; phenotype; polymerase chain reaction; prostate neoplasms; protein purification; protein sequence; secretory protein; transforming growth factors; tumor suppressor genes; western blottings

Inductive stimuli from mesenchymal cells direct the growth and phenotypic differentiation of epithelium in prostate gland development. This process is androgen regulated, yet specific mechanisms are unknown. To address mechanisms, we have characterized paracrine acting factors from fetal rat urogenital sinus (UGS) organ cultures and derived mesenchymal cell lines. UGS mesenchymal cells secrete a factor (UGIF) which alters epithelial cell phenotype, stimulates secretory protein synthesis, and inhibits proliferation, suggesting this protein functions as a differentiation-inducing factor. Physicochemical, biological, and immunological properties have been determined and the UGIF protein purified to homogeneity. UGIF inhibits cell proliferation, stimulates protein secretion, and alters phenotype in PC-3 and LNCaP human prostatic carcinoma cell lines, suggesting a tumor suppressor function. A variant, phenotypically transformed UGS mesenchymal cell strain showed decreased UGIF expression coordinate with a stimulated secretion of activated TGF-B2 not observed in the parent cell line. Mutations or alterations in UGIF expression coupled with alterations in other growth factors may produce a growth factor imbalance, potentially relevant to the initiation and progression of prostatic cancer. The following studies are proposed: To develop antibody and oligonucleotide probes to UGIF; To determine the spatial and temporal expression of UGIF and TGF-B isoforms in human normal prostate and in clinical stages of human prostatic cancer; To determine the primary structure of human UGIF cDNA and characterize mutational defects in prostate cancer by analysis of PCR amplified cDNA products encoding UGIF and; To characterize the in vitro and in vivo responses of PC-3 and LNCaP human prostatic adenocarcinoma cells to UGIF. Antibody and cRNA probes will be used for immunocytochemistry and in situ hybridization. The mixed oligonucleotide primed amplification procedure (MOPAC) will used to analyze human UGIF cDNA, generate probes, and clone full length cDNA. UGIF cDNA from stages of human prostatic carcinoma will be analyzed by PCR/GC-clamp DGGE methods and sequenced to map putative mutations. PC-3 and LNCaP cell exposed to purified rat UGIF +/- androgens will be analyzed for growth and marker protein expression in vitro to correlate with in vivo effects. The athymic nude mouse host system will be used with PC-3 and LNCaP cells inoculated with UGIF secreting cells or UGIF containing Alzet Micro-Osmotic pumps to assess in vivo effects. Long range goals are to understand the specific mechanisms of UGIF action in prostate neoplasia and the potential therapeutic value of UGIF as a prostate tumor suppressor.

间充质细胞的感应刺激指导生长和 前列腺发育中上皮的表型分化。 此过程是受雄激素调节的，但特定机制尚不清楚。 为了解决机制，我们表征了旁分泌作用因子 来自胎儿大鼠泌尿生殖器窦（UGS）器官培养物并得出 间充质细胞系。 UGS间充质细胞分泌一个因子（UGIF） 哪个改变上皮细胞表型，刺激分泌蛋白 合成并抑制增殖，表明该蛋白质功能 作为差异化因素。 物理化学，生物学和 已经确定了免疫学特性，UGIF蛋白 纯化为同质性。 UGIF抑制细胞增殖，刺激 蛋白质分泌，并在PC-3和LNCAP人类前列腺中改变表型 癌细胞系，表明肿瘤抑制功能。 一个变体 表型转化的UGS间充质细胞菌株显示降低 UGIF表达与激活的刺激分泌坐标 在母细胞系中未观察到TGF-B2。 突变或改变 UGIF表达与其他生长因子的变化相结合可能 产生生长因子不平衡，可能与开始有关 和前列腺癌的进展。 提出了以下研究： 为UGIF开发抗体和寡核苷酸探针；确定 人类中UGIF和TGF-B同工型的空间和时间表达 正常前列腺和人类前列腺癌的临床阶段；到 确定人UGIF cDNA的主要结构并表征 通过分析PCR扩增cDNA的前列腺癌突变缺陷 编码UGIF的产品；表征体内和体内 PC-3和LNCAP人前列腺腺癌细胞对UGIF的反应。 抗体和CRNA探针将用于免疫细胞化学和原位 杂交。 混合寡核苷酸引发的扩增过程 （MOPAC）将用于分析人类UGIF cDNA，生成探针和克隆 全长cDNA。 人类前列腺癌阶段的UGIF cDNA 将通过PCR/GC-CLAMP DGGE方法分析，并测序用于MAP 推定的突变。 PC-3和LNCAP细胞暴露于纯化的大鼠UGIF +/- 将分析雄激素的生长和标记蛋白表达 体外与体内效应相关。 无胸腺裸鼠宿主 系统将与接种UGIF接种的PC-3和LNCAP单元 分泌细胞或含有Alzet微渗透泵评估的细胞或UGIF 体内效应。 远距离目标是了解特定机制 前列腺肿瘤和潜在治疗价值的UGIF作用 UGIF作为前列腺肿瘤抑制剂。