HUMAN PROSTATE DERIVED GROWTH FACTOR

人前列腺源性生长因子

• 批准号: 3198361
• 负责人: J. W. none CHIAO
• 金额: $ 17.68万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-07-01 至 1994-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3198361
• 关键词: DNA replication; androgens; benign prostate hyperplasia; cell death; cell growth regulation; complementary DNA; electrofocusing; enzyme linked immunosorbent assay; fibroblast growth factor; gel filtration chromatography; growth factor; growth factor receptors; high performance liquid chromatography; human tissue; immunoperoxidase; ion exchange chromatography; laboratory mouse; messenger RNA; molecular cloning; monoclonal antibody; nucleic acid sequence; prostate disorder; prostate neoplasms; protein purification; ribosomal RNA

We have described a prostatic cell derived growth factor (PRGF) from a human prostatic epithelial cancer cell line JCA-1. PRGF is a protein, constitutively produced by JCA-1 cells and released into the culture medium. PRGF was purified and shown to have a molecular weight of 53,100. A partial amino acid sequence from the N-terminal of PRGF has been obtained. The interaction between stromal fibroblast preparations, from human prostate or nonprostate fibroblasts, and PRGF causes an accelerated growth of the fibroblasts. The mechanism includes an increase in growth rate and a more rapid entry of G1 phase cells into the replicating S-phase of the cell cycle. PRGF may represent a paracrine growth factor and is distinct from the commonly known growth factors. Since prostate growth involves both epithelium and fibromuscular tissue, and prostate hyperplasia and malignancy are sometimes characterized by abnormal fibroblast- epithelial cell interactions, we plan to investigate the nature and biological activity of the PRGF. We will further characterize and identify PRGF from JCA-1 cancer cell culture supernatant. PRGF needs to be characterized to further identify its amino acid sequence and production of polyclonal and monoclonal antibodies to investigate its biological functions. Sensitive assay procedures, such as cell cycle analysis, cell size, DNA replication, etc. will be developed to identify PRGF. The biological activity of PRGF will be pursued by identifying the cell types in the prostate, whether stromal or epithelial, responding to PRGF. Growth responses of different cell types will be measured and cell surface receptors, specific for PRGF, will be identified using radiolabeled PRGF. Localization of the PRGF production site in clinical prostate tissues will be pursued using the peroxidase antibody technique. Normal, benign and cancer tissues from the prostate will be analyzed in order to correlate PRGF and prostate cancer gradings. Furthermore, the relationship between PRGF production and androgen stimulation will be analyzed using JCA-1 cells. Further structural analysis of PRGF, using isolated JCA-1 cDNA and PRGF nucleotide sequence is planned. The cDNA clone will also make it possible to heterologously express the PRGF protein.

我们描述了一种来自前列腺细胞的生长因子（PRGF） 人前列腺上皮癌细胞系JCA-1。 PRGF是一种蛋白质， 由 JCA-1 细胞组成型产生并释放到培养物中 中等的。 PRGF 经纯化后显示分子量为 53,100。 PRGF N 末端的部分氨基酸序列已被 获得。 基质成纤维细胞制剂之间的相互作用，来自 人前列腺或非前列腺成纤维细胞，PRGF 会导致加速 成纤维细胞的生长。 该机制包括增加生长 率和 G1 期细胞更快进入复制 S 期 细胞周期。 PRGF可能代表一种旁分泌生长因子并且是 与众所周知的生长因子不同。 由于前列腺生长 涉及上皮和纤维肌肉组织，以及前列腺增生 恶性肿瘤有时以成纤维细胞异常为特征 上皮细胞相互作用，我们计划研究其性质和 PRGF 的生物活性。 我们将进一步表征和鉴定 JCA-1 癌细胞中的 PRGF 培养上清液。 PRGF 需要进行表征以进一步识别 其氨基酸序列以及多克隆和单克隆的产生 抗体来研究其生物学功能。 灵敏检测 程序，例如细胞周期分析、细胞大小、DNA 复制等。 将被开发来识别PRGF。 PRGF的生物活性将 通过识别前列腺中的细胞类型（无论是间质细胞）来追求 或上皮细胞，对 PRGF 做出反应。 不同细胞的生长反应 将测量类型，并且针对 PRGF 的细胞表面受体将 使用放射性标记的 PRGF 进行鉴定。 PRGF生产本地化 将使用过氧化物酶追踪临床前列腺组织中的位点 抗体技术。 前列腺的正常、良性和癌组织 将进行分析，以便将 PRGF 和前列腺癌分级相关联。 此外，PRGF 产生与雄激素之间的关系 将使用 JCA-1 细胞分析刺激。 进一步的结构 使用分离的 JCA-1 cDNA 和 PRGF 核苷酸序列分析 PRGF 计划。 cDNA克隆也将使得异源性成为可能 表达 PRGF 蛋白。