Molecular characterization of endometrial cancer

初步数据确定了子宫内膜癌细胞系亚群中NF-κ B信号传导的需要。与G.L.麦克斯韦合作，我们检测了子宫内膜癌的基因表达谱中NF-κ B活性的特征，其在浆液性与类浆液性组织学中平衡，并且在非裔美国人（AA）和高加索人（C）妇女中均匀分布。我们发现，签名是显着较高的癌症从AA妇女，特别是那些浆液性组织学变异。我们实验室正在进行的项目，以及与麦克斯韦实验室的合作，将继续解决既定的目标。我们通过用IKKb小分子抑制剂处理9种子宫内膜癌细胞系并在XTT测定中定量存活率来测试它们对NF-κ B信号传导的依赖性。我们将这些实验扩展到其他细胞系，并进行了其他功能测定，如粘附、侵袭和细胞因子产生。我们在体外鉴定了至少2种对IKKb抑制敏感的子宫内膜癌细胞系。我们分析了在使用小分子抑制剂阻断IKKb信号传导和引入作为IKKb介导的信号传导的“超级阻遏物”的IkBa突变形式后这2个细胞系中的基因表达变化。我们通过蛋白质印迹证实了子宫内膜癌特异性NF-κ B靶蛋白表达的降低。这些基因和蛋白质特征将使我们能够使用我们的合作者目前正在收集和生成的基因表达谱和组织微阵列来特异性地询问子宫内膜癌样本。这些研究的总体结果将是设计NF-kB靶向药物的临床试验，用于子宫内膜癌显示通路激活证据的妇女。

Preliminary data established the requirement for NF-kB signaling in a subset of endometrial cancer cell lines. In collaboration with G.L.Maxwell, we examined a signature of NF-kB activity in the gene expression profiles of endometrial cancers balanced for serous vs endometrioid histology and also distributed equally among African American (AA) and caucasian (C) women. We found that the signature was significantly higher in cancers from AA women, especially those of the serous histology variant. Ongoing projects in our lab, and in collaboration with the Maxwell lab, will continue to address the stated goals. We tested 9 endometrial cancer cell lines for their dependence on NF-kB signaling by treating them with IKKb small molecule inhibitor and quantifying viability in XTT assays. We extended these experiments to additional cell lines, and performed additional function assays such as adhesion, invasion and cytokine production. We identified at least 2 endometrial cancer cell lines that are sensitive to IKKb inhibition in vitro. We profiled gene expression changes in these 2 cell lines upon blockade of IKKb signaling using both a small molecule inhibitor and introduction of a mutant form of IkBa that serves as a "super-repressor" of IKKb-mediated signaling. We confirmed decrease in protein expression of the endometrial cancer-specific NF-kB targets by Western blot. These gene and protein signatures will allow us to specifically interrogate the endometrial cancer samples using gene expression profiles and tissue microarrays that are currently being collected and generated by our collaborator. The overall outcome of these studies will be to design clinical trials with NF-kB targeted agents for women whose endometrial cancers show evidence of pathway activation.