Development of monitoring system of colorectal cancer

本研究的目的是利用课题组新发现的癌细胞特异性表达物质Bradeion开发结直肠癌的诊断技术，并最终开发包括癌细胞靶向药物在内的新的治疗技术。2003年，我们通过两个独立的系统证明了Bradeion分子（一个基因和一个蛋白质）用于结直肠癌诊断技术的实用性；实时RT-PCR检测表达的Bradeion基因（专利申请期间），免疫层析（试纸条法）检测单克隆抗体标本的Bradeion蛋白（参考文献4）。2004年，一种新的简单的测量GTPase活性的方法被开发出来（文献1），并证明了Bradeion是GTPase，是加速癌细胞细胞分裂的能量供应系统的中心。我们还通过所描述的技术（文献3）继承了Bradeion的结晶，用于蛋白质的结构分析（文献2）。现在我们正在对晶体进行结构分析，并寻找一种活性位点特异性抑制剂。此外，我们还制备了针对本小组鉴定的Bradeion GTPase的单克隆抗体。我们在培养的结直肠癌细胞和患者组织中使用针对Bradeion蛋白全区或GTPase的单克隆抗体进行免疫组化，研究肿瘤特异性和位置。我们设计了针对Bradeion核苷酸序列的RNAi，采用RNA干扰技术作为治疗技术之一，并将其引入培养的结直肠癌细胞中，观察对其的影响。现在我们正在研究用腺病毒载体对它们的影响。

A purpose of this study is to develop diagnostic technology of colorectal cancer with a carcinoma cell-specific expressed material, Bradeion discovered by our research group newly, and to finally develop new treatment technology including a cancer cell-targeting medicine.In 2003, we demonstrated utility of a Bradeion molecule (a gene and a protein) for a diagnostic technology of a colorectal carcinoma by two independent system; real time RT-PCR to measure the expressed Bradeion gene (during a patent application), and an immunochromatography (test strip method) to detect Bradeion protein of specimens with a monoclonal antibodies (ref. 4).In 2004, a new simple method measuring the GTPase activity is developed (ref. 1), and proved that Bradeion is GTPase, a center of the energy supply system to enable accelerated cell division of a cancer cell. We also succeeded the crystallization of Bradeion for structure analysis of the protein (ref. 2) by the technology described (ref 3). Now we are performing structure analysis of the crystal, and searching for an active site-specific inhibitor. Furthermore, we made monoclonal antibodies against the GTPase of Bradeion identified by our group. We performed immunohistochemistry using the monoclonal antibodies against the whole region or GTPase of Bradeion protein in cultured colorectal carcinoma cells and patients' tissue and studied cancer-specificity and location. We designed the RNAi specific to the nucleotide sequence of Bradeion, using RNA interference technology for as one of the therapy technology, and introduced it into cultured colorectal carcinoma cells and examined influence to them. Now we are investigating influence to them with an adenovirus vector.