CYTOGENETIC ANALYSIS OF HUMAN PROSTATE CANCER

赠款申请的总体目的是确定遗传和 与肿瘤发生和转移相关的生物学参数 恶性前列腺癌 肿瘤组织将从 非裔美国人（75%）和高加索人（25%）患者（约20至 30/年），在肿瘤进展的早期和晚期Gleason阶段。 细胞将 通过皮下（s.c.）植入 严重联合免疫缺陷（SCID）小鼠中的基质胶。 男性和 女性SCID将用于尝试分离雄激素依赖性和 独立的肿瘤变体。 原发性肿瘤细胞系将从 南卡罗来纳利用我们实验室开发的程序。 第一个目标是基于其对原发性肿瘤系的分类。 在“Boyden趋化性测定”中的侵袭能力。侵入性和 将非侵入性亚系与原系分离并维持 低通道（小于5）。 此外，将对SCID小鼠i. v. 与侵袭性细胞，以确定转移的潜力， 亚系，并从发生的任何转移中获得培养物。 的 第二个目标是对原发性肿瘤系进行核型分析，非侵入性的， 侵袭和转移亚系。 通过广泛的核型分析， 通过比较，应该可以确定一致的染色体 在肿瘤发生的早期阶段， 培养物的侵袭和/或转移能力。 的延伸 染色体组型工作，第三个目标是使用流式细胞术来测量DNA 变异体的含量和非整倍性。 原位杂交和 荧光显微镜与cDNA探针的染色体（i. e. , 染色体2、7、10、13、15、16、17和Y）将用于评估 在不同的亚系中是否获得或丢失了特定的染色体。 流式细胞术单变量分析染色体含量 中期细胞将有助于确定是否缺失或总易位 发生. 第四个目标是检查肿瘤细胞系的扩增和表达 癌基因的（即，c-myc、H-ras、Ki-ras、fos） 印迹分析。 此外，原位杂交将显示癌基因 扩增发生在均匀染色的区域，或者如果癌基因 与大易位、标记染色体或双 分钟 综合起来，这些实验有助于确定特定的 与前列腺癌有关的染色体区域。 任何的一致性 将通过评价培养物来确定观察到的染色体变化 从各种各样的病人中建立起来的。 “数据管理器”将提供 患者的年龄、病史、种族、治疗和阶段信息 癌症的进展。

The overall purpose of the grant application is to define the genetic and biological parameters associated with the initiation and metastases of malignant prostate cancer. Tumor tissue will be obtained from Afro-American (75%) and caucasian (25%) patients (approximately 20 to 30/yr) at early and late Gleason stages of tumor progression. Cells will be grown up for culturing by subcutaneous (s.c.) implantation with Matrigel in severe combined immune deficient (SCID) mice. Both male and female SCIDs will be used in attempts to isolate androgen dependent and independent tumor variants. Primary tumor lines will be established from the s.c. tumor tissue utilizing procedures developed in our laboratory. The first goal is to classify the primary tumor lines based on their invasive ability in 'Boyden chemotactic assays' . Invasive and non-invasive sublines will be isolated from primary lines and maintained at low passage (less than 5). Further, SCID mice will be injected i.v. with the invasive cells to determine the metastatic potential of the sublines and to obtain cultures from any metastases that occur. The second goal is to karyotype the primary tumor lines, the non-invasive, the invasive and the metastatic sublines. By extensive karyotyping and comparison it should be possible to identify consistent chromosomal aberrations at early stages in tumorigenesis and perhaps in relation to the invasive and/or metastatic abilities of the culture. In extension of the karyotype work, a third goal is to use flow cytometry to measure DNA content and aneuploidy of the variants. In situ hybridization and fluorescence microscopy with cDNA probes to chromosomes (i. e. , chromosomes 2, 7, 10, 13, 15, 16, 17 and Y) will be used to assess whether specific chromosomes are gained or lost in the various sublines. Flow cytometry with univariate analysis of the chromosomal content of metaphase cells will help determine if deletions or gross translocations occur. A fourth goal, is to examine tumor lines for amplification and expression of oncogenes (i.e., c-myc, H-ras, Ki-ras, fos) by Southern and Northern blot analyses. In addition, in situ hybridization will show if oncogene amplification occurs in homogeneously staining regions or if oncogenes are associated with gross translocations, marker chromosomes or double minutes. Taken together, these experiments help identify specific chromosomal regions involved in prostate cancer. The consistency of any chromosomal changes observed will be determined by evaluation of cultures established from a variety of patients. The 'data manager' will provide information on the patients age, history, race, treatment and stage of progression of the cancer.