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来挖掘可通过 NCIS癌症基因组解剖项目，我们确定了665个 这些序列在结肠癌文库中表达，但在必需的正常组织中不表达。 其中，356个是在Affytext表达式数组上表示的，这允许 这些基因在结肠癌组织和细胞系中表达的比较，以及 重要的正常组织这项调查加上随后的确认， 定量RT-PCR鉴定了两个以前未表征的基因 其表达相对特异于相当大比例的结肠癌样品 测试.我们进行了实验，以确定siRNA是否介导了 抑制我们的候选基因抑制HCT 15结肠癌细胞的细胞增殖 细胞增殖测定使用Cyquant试剂盒抑制细胞 与siRNA阴性对照相比，细胞增殖减少10%。siRNA介导的沉默 这些基因导致HCT 15细胞系中细胞增殖的少量减少，表明 这些基因可能在促进结肠癌细胞增殖中发挥作用。额外 实验表明IGFL2在MAP激酶的调节中起着假定的作用 通路随后的研究涉及在保守区的定点诱变， IGFL2将进行，以确定这些地区的结构作用， 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来挖掘可通过 NCIS癌症基因组解剖项目，我们确定了665个 这些序列在结肠癌文库中表达，但在必需的正常组织中不表达。 其中，356个是在Affytext表达式数组上表示的，这允许 这些基因在结肠癌组织和细胞系中表达的比较，以及 重要的正常组织这项调查加上随后的确认， 定量RT-PCR鉴定了两个以前未表征的基因 其表达相对特异于相当大比例的结肠癌样品 测试.我们进行了实验，以确定siRNA是否介导了 抑制我们的候选基因抑制HCT 15结肠癌细胞的细胞增殖 细胞增殖测定使用Cyquant试剂盒抑制细胞 与siRNA阴性对照相比，细胞增殖减少10%。siRNA介导的沉默 这些基因导致HCT 15细胞系中细胞增殖的少量减少，表明 这些基因可能在促进结肠癌细胞增殖中发挥作用。额外 实验表明IGFL2在MAP激酶的调节中起着假定的作用 通路随后的研究涉及在保守区的定点诱变， IGFL2将进行，以确定这些地区的结构作用， MAP激酶的激活。

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 NCIs 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. Additional experiments revealed that IGFL2 plays a putative role in the regulation of the MAP Kinase pathway. Subsequent studies involving site directed mutagenesis in the conserved regions of the IGFL2 will be carried out in order to determine the structural role of these regions in the activation of MAP Kinases.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 NCIs 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. Additional experiments revealed that IGFL2 plays a putative role in the regulation of the MAP Kinase pathway. Subsequent studies involving site directed mutagenesis in the conserved regions of the IGFL2 will be carried out in order to determine the structural role of these regions in the activation of MAP Kinases.