DETECTION OF MICROMETASTATIC DISEASE BY RT-PCR

临床局限性前列腺癌根治术（RP）后 (CaP)15-40%的男性会表现出疾病的临床或生化证据 进展许多这些失败是由于隐匿性转移性疾病。 这个项目探讨了前列腺癌扩散的现象学 通过应用逆转录聚合酶链反应， （RT-PCR）。该项目的具体目标是： 1)为了验证前列腺外肿瘤细胞存在于 大多数具有临床局限性CaP但在RP后清除的患者 这些患者治愈了CaP，但没有清除或后来再次出现， 复发性疾病发展。 2)检查骨髓中RT-PCR结果的现象学 前列腺粒子植入（PSI）后，并将结果与 RP之后的人。 3)检测骨髓中CaP标志物、hK 2和PSMA的RT-PCR方法 标本，并检查各种处理（RP，辐射， 激素）对这些标志物单独和与 PSA。 4)为了评估新的RT-PCR检测毒力标记物， 技术核心。 5)为了测试新的半定量基因表达分析， 随着疾病严重程度的增加， 前列腺癌的发病率会增加。 将对术前和术后骨髓穿刺进行连续检查。 从接受根治性直肠切除术的患者中收集。标本将 用RT-PCR测定法分析PSA表达。结果将 与长期（5年）无病生存率和生化指标相比， 复发，如通过生物发光测定所定义的， 检测限为0.008 ng/ml。也将进行类似的研究 接受PSI的患者。来自RP和PSI患者的库化cDNA将 也可以通过技术核心中描述的RT-PCR检测进行检测， 帮助评估骨髓细胞检测的转移潜力， 也将被研究。最后，我们将研究转录介导的 放大是否半定量技术，这将使我们能够 了解PSA、PSMA和hK 2的相对表达，提供额外的 超出当前定性分析提供的信息。

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.