DETECTION OF MICROMETASTATIC DISEASE BY RT-PCR

临床局限性前列腺癌根治性切除术(RP)后 (上限)15%-40%的男性会出现疾病的临床或生化证据 进步。这些失败中的许多是由于隐匿性的转移性疾病。 这个项目探索前列腺癌扩散的现象学。 逆转录聚合酶链式反应的应用 (RT-PCR)。该项目的具体目标是： 1)检验前列腺外肿瘤细胞存在于 大多数临床上有局限性CAP的患者在接受RP治疗后仍有明确的症状 那些治愈了CAP，但没有清除或稍后再次出现的患者 疾病会反复发作。 2)检查骨髓中RT-PCR结果的现象学 前列腺粒子植入(PSI)后的结果并与 那些跟在RP后面的。 3)采用RT-PCR法检测骨髓CAP标志物hk2和psMA。 并检查不同处理(放射治疗、放射治疗、 激素)单独作用于这些标志物的表达，并与 PSA。 4)评估新的RT-PCR检测毒力标记的方法，该方法由 技术核心。 5)检验新的半定量基因表达检验假说 随着疾病严重程度的增加，表达基因的细胞数量增加 与前列腺癌相关的人数将会增加。 术前和术后连续的骨髓抽吸物将 收集自接受根治性前列腺切除术的患者。标本将会 用RT-PCR法分析PSA的表达。结果将是 与长期(5年)无病生存和生化指标比较 用生物化学发光分析法定义的复发 检出限为0.008 ng/ml。类似的研究也将在 接受PSI的患者。来自RP和PSI患者的文库cDNA会 也可以通过技术核心中描述的RT-PCR分析进行测试，该技术核心可以 帮助评估骨髓中细胞检测的转移潜力， 也将进行研究。最后，我们将用转录介导法进行检查 放大是否采用半定量技术，这将使我们能够 了解PSA、PSMA和HK2的相对表达，提供其他 目前的定性分析所提供的信息之外的信息。

After radical prostatectomy (RP) for clinically localized prostate cancer (CaP) 15-40% of men will show clinical or biochemical evidence of disease progression. Many of these failures are due to occult metastatic disease. This project explores the phenomenology of prostate cancer dissemination through application of reverse transcriptase polymerase chain reaction (RT-PCR). The specific aims of this project are: 1) To test the hypothesis that extra-prostatic tumor cells are present in most patients with clinically localized CaP yet clear following RP in those patients cured of CaP, but do not clear or later reappear when recurrent disease develops. 2) To examine the phenomenology of RT-PCR results in the bone marrow following prostate seed implantation (PSI) and compare the results with those following RP. 3) To test RT-PCR assays for the CaP markers, hK2 and PSMA in bone marrow specimens and examine the effect of various treatments (RP, radiation, Hormones) on the expression of these markers alone and in combination with PSA. 4) To assess new RT-PCR assays for markers of virulence developed by the technical core. 5) To test new semi-quantitative gene expression assays the hypothesis that as disease severity increases the number of cells expressing genes associated with prostate cancer will increase. Pre-operative and serial post-operative bone marrow aspirates will be collected from patients undergoing radical prostatectomy. Specimens will be analyzed with an RT-PCR assay for PSA expression. Results will be compared with long-term (5 year) disease free survival and biochemical recurrence as defined by a chemiluminescent assays with a biological detection limit of 0.008 ng/ml. A similar study will also be performed on patients undergoing PSI. The banked cDNA from the RP and PSI patients will also be tested by RT-PCR assays described in the Technical Core, which may help assess the metastatic potential of cell detection in the bone marrow, will also be studied. Finally, we will examine with transcription mediated amplification whether semi-quantitative techniques, which will allow us to know the relative expression of PSA, PSMA, and hK2, provide additional information beyond that provided by the current qualitative assays.