Molecular characterization of endometrial cancer

初步数据确定了子宫内膜癌细胞系中对核因子-kB信号的需求。与G.L.Maxwell，CCR，WRAMC合作，我们计划检查子宫内膜癌基因表达谱中核因子-kB活性的特征，在浆液性和子宫内膜样癌组织学中平衡，也在非裔美国人(AA)和高加索人(C)女性中平均分布。在AA妇女的癌症中，特别是那些浆液性组织学变异的癌症中，初步特征明显更高。我们的实验室正在进行的项目，以及与Maxwell实验室的合作，将继续实现上述目标。我们测试了9个子宫内膜癌细胞株对NF-kB信号的依赖性，方法是用IKKB小分子抑制剂处理它们，并在XTT试验中定量测定它们的活力。我们计划将这些实验扩展到更多的细胞系，并进行更多的功能测试，如黏附、侵袭和细胞因子的产生。我们在体外鉴定了至少2个对IKKB抑制敏感的子宫内膜癌细胞株。我们分析了在使用小分子抑制剂和引入突变形式的IkBA作为IKKB介导的信号的“超级抑制子”来阻断IKKB信号时，这两种细胞系的基因表达变化。我们将通过Western印迹来证实子宫内膜癌特异性核因子-kB靶点的蛋白表达降低。这些基因和蛋白质签名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定地询问子宫内膜癌样本。这些研究的总体结果将是为子宫内膜癌显示通路激活的证据的妇女设计针对核因子-kB靶向药物的临床试验。

Preliminary data established the requirement for NF-kB signaling in a subset of endometrial cancer cell lines. In collaboration with G.L.Maxwell, CCR, WRAMC, we plan to examine 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. A preliminary 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 plan to extend these experiments to additional cell lines, and in 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 will confirm 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.