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。在与G.L.Maxwell的合作中，我们检查了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上是平衡的，在非裔美国人(AA)和高加索人(C)女性中也是平均分布的。我们发现，在AA女性的癌症中，这种签名明显更高，特别是那些浆液性组织学变异的癌症。我们实验室正在进行的项目，并与麦克斯韦尔实验室合作，将继续实现所述目标。我们检测了9个子宫内膜癌细胞系对核因子-kB信号的依赖性，方法是用IKKB小分子抑制剂处理它们，并用XTT法测定它们的存活率。我们将这些实验扩展到更多的细胞系，并进行了额外的功能测试，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。我们通过Western印迹证实子宫内膜癌特异性核因子-kB靶点的蛋白表达减少。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具体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表现出途径激活的证据的妇女设计以核因子-kB为靶向的药物的临床试验。

Preliminary data established the requirement for NF-kB signaling in a subset of endometrial cancer cell lines. In collaboration with G.L.Maxwell, 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.