Development and standardization of new molecular diagnostic methods for genito-urinary cancers

本研究的目的是在早期发现、疾病进展预测和选择对化疗有反应的合适患者方面开发新的生殖-泌尿系统癌分子诊断方法。使所开发的方法标准化也是一个重要目标。在早期检测方面，我们发现尿中CD44v8-10与标准CD44 mRNA的比值是膀胱癌早期检测的有用指标。我们还发现MMP-9和TIMP-2在浅表性膀胱癌中的表达分别是膀胱内复发的独立预测因子。在预测预后方面，膀胱癌的侵袭性与CD44v8-10与标准CD44的比值及纤溶酶原激活物的表达相关。通过基因谱分析，我们发现了一个新的基因集，可以预测肾癌患者的预后。我们还发现血清中VEGF、HGF和内皮抑素的浓度是肾癌的有效预后指标。我们开发并标准化了一种巢式RT-PCR检测尿路上皮癌患者外周血中uroplakin II mRNA的方法。该方法有望用于微转移的检测。在预测阶梯癌患者对化疗的良好反应方面，酵母功能检测评估的p53功能异常与MRP1（多药耐药基因成员）的表达升高相关。膀胱癌患者顺铂联合化疗后MDR1、MRP1、2、3基因表达升高。此外，在膀胱癌组织中，阿霉素耐药与MDR1和MRP1相关。

The aim of this study was to develop new molecular diagnostic methods for genito-urinary cancers in terms of early detection, prediction of disease progression, and selection of proper patients who will respond to chemotherapy. Standardization of the developed method was also an important goal. As to early detection, we found that the mRNA ratio of CD44v8-10 to standard from of CD44 in urine was a useful marker for early detection of bladder cancer. We also found that MMP-9 and TIMP-2 expression in superficial bladder cancer were respectively independent predictors for intravesical recurrence. As to prediction of prognosis, invasiveness of bladder cancer was correlated with the ratio of CD44v8-10 to standard form of CD44 and plasminogen activator expression. By the gene profiling analysis using microarray, we found a novel gene set that can predict prognosis of kidney cancer patients. We also identified that serum concentration of VEGF, HGF, and endostatin was useful prognostic marker for kidney cancer. We developed and standardized a method of nested RT-PCR for uroplakin II mRNA in peripheral blood of urothelial cancer patients. The method was assumed to be promising as to detection of micrometastasis. As to prediction of good responders to chemotherapy in ladder cancer patients, aberration of p53 function evaluated by the yeast functional assay was correlated with elevated expression of MRP1 (a member of multi-drug resistance genes). Expression of MDR1 and MRP1, 2, 3 genes was increased after cisplatin-based combination chemotherapy in bladder cancer patient. Moreover, resistance to adriamycin was correlated with MDR1 and MRP1 and in bladder cancer tissues.