CYTOGENETIC ANALYSIS OF HUMAN PROSTATE CANCER

赠款申请的总体目的是定义遗传和 与卵巢癌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。单元格 经皮下(S.C.)培养而成熟植入 严重联合免疫缺陷(SCID)小鼠的Matrigel。无论是男性还是 女性SCID将用于分离雄激素依赖和 独立的肿瘤变种。原发肿瘤细胞系将从 南卡罗来纳州我们实验室开发的肿瘤组织利用程序。 第一个目标是根据它们的类型对原发肿瘤系进行分类 《博伊登趋化试验》中的侵袭能力。侵入性和 非侵入性次级线路将与主要线路隔离并进行维护 在低通道时(小于5)。此外，SCID小鼠将被静脉注射。 用侵袭性细胞来确定肿瘤的转移潜能 并从发生的任何转移中获得培养物。这个 第二个目标是对原发肿瘤系，非侵袭性， 侵袭性和转移性亚系。通过广泛的核型测定和 比较一下，应该可以识别出一致的染色体 肿瘤发生的早期阶段的异常，可能与 培养物的侵袭和/或转移能力。在扩展中 核型工作的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体。原位杂交和 带有对染色体的cDNA探针的荧光显微镜(即， 染色体2、7、10、13、15、16、17和Y)将用于评估 特定的染色体是否在不同的亚系中获得或丢失。 流式细胞术在单变量分析中的应用 中期细胞将有助于确定缺失或严重易位 发生。 第四个目标是检查肿瘤细胞系的扩增和表达。 南方和北方的癌基因(即c-myc、H-ras、Ki-ras、fos) 印迹分析。此外，原位杂交将显示癌基因是否 扩增发生在均匀染色的区域或癌基因 与严重易位、标记染色体或双染色体有关 几分钟。综上所述，这些实验有助于识别特定的 与前列腺癌有关的染色体区域。任一项的一致性 观察到的染色体变化将通过对培养物的评估来确定 从各种各样的病人中建立起来的。数据经理将提供 关于患者的年龄、病史、种族、治疗和分期的信息 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.