Molecular Targets - Colon Cancer

我们采用了一种新的策略来鉴定治疗的分子靶点， 预防常见癌症，最初重点是结肠癌。的一个主要 化疗剂的局限性是相对狭窄的治疗：毒性比， 这是由于癌性和正常组织中治疗靶点的共同性 组织中因此，我们试图确定潜在的治疗靶点， 通过使用公共数据库中可获得的基因表达信息， 使用高通量基因表达阵列分析。使用数字基因 expression Displayer，用于挖掘通过NCI的Cancer 基因组解剖计划，我们确定了665个序列，在结肠癌中表达 文库中，但不是在必需的正常组织中。其中356人在Affyoung有代表 表达阵列，这允许比较这些基因在结肠中的表达 癌组织和细胞系以及必需的正常组织。这次调查加上 随后使用定量RT-PCR进行确认， 以前未表征的基因，其表达相对特异于实质性的 结肠癌样本的检测比例。我们进行了实验，以确定 siRNA介导的对候选基因的抑制是否会抑制细胞增殖， HCT 15结肠癌细胞系;使用Cyquant 与siRNA阴性对照相比，试剂盒抑制细胞增殖10%。siRNA 这些基因介导的沉默导致HCT 15中细胞增殖的小幅降低 细胞系，表明这些基因可能在促进细胞增殖中起作用， 结肠癌使用可用数据库和高吞吐量的方法 用于鉴定相对癌症选择性基因靶标的技术可构成 一种快速有效的开发活性和毒性较小的化疗药物的方法， 剂.这些基因作为潜在靶点的鉴定完全基于其 表达谱，而不是依赖于其细胞内的先验知识， function（s）.除了为药物开发提供感兴趣的靶标之外， 也可以用于药物输送，作为疾病的分子标记物，或作为替代物， 用于药物递送和药物有效性的标记物。

We have employed a novel strategy to identify molecular targets for the therapy and prevention of common cancers, with an initial focus on colon cancer. One of the major limitations of chemotherapeutic agents is the relatively narrow therapeutic: toxic ratio that results from the commonality of the therapeutic target in both the cancerous and normal tissues. Therefore, we sought to identify potential therapeutic targets that were relatively cancer specific by using gene expression information available in public databases coupled with the use of high-throughput gene expression array analysis. Using the Digital Gene expression Displayer to mine expression libraries available through the NCI's Cancer Genome Anatomy Project, we identified 665 sequences that were expressed in colon cancer libraries but not in essential normal tissues. Of these, 356 were represented on Affymetrix expression arrays and this allowed for a comparison of the expression of these genes in colon cancer tissues and cell lines, and essential normal tissues. This investigation coupled with subsequent confirmation using quantitative RT-PCR resulted in the identification of two previously uncharacterized genes whose expression was relatively specific to a substantial proportion of colon cancer samples tested. We carried out experiments in order to determine whether the siRNA mediated suppression of our candidate genes inhibits cell proliferation in the HCT15 colon cancer cell line; cell proliferation assays were carried out using a Cyquant kit inhibit cell proliferation by 10% as compared to the siRNA negative control. siRNA mediated silencing of these gene resulted in a small reduction of cell proliferation in HCT15 cell lines indicating that these genes may play a role in promoting cell proliferation in colon cancer. The approach of using available databases coupled with high-throughput technologies for the identification of relatively cancer selective gene targets may constitute a rapid and efficient method for the development of active and less toxic chemotherapeutic agents. The identification of these genes as potential targets was based entirely on their expression profile rather than dependent on prior knowledge of their intracellular function(s). In addition to providing targets of interest for drug development, such targets could also be exploited for drug delivery, as molecular markers of disease, or as surrogate markers for drug delivery and drug effectiveness.