Role of the human transcription factor T in chordoma growth

在这个项目的第二年，我们执行了必要的材料转让协议，以获得已公布的脊索瘤细胞系，包括新的脊索瘤细胞系JHC-1和MUG-Chor1。在这四个细胞系中，通过免疫印迹和免疫沉淀，我们证实了人类T的高水平表达。实验表明，抗T shRNA处理显著减缓了脊索瘤细胞系的增殖。适当的检测证实了T基因和蛋白的下调。基因芯片分析是比较抗T shRNA处理的细胞株和对照shRNA处理的细胞株目前正在进行中，预计在一个月内完成。此外，还对脊索瘤细胞的基础microRNA环境进行了分析。最后，我们还鉴定了脊索瘤细胞系在电离辐射处理后表达发生变化的microRNA。对类似实验的cDNA微阵列分析显示，许多已知的、受电离辐射调节的靶向基因。验证性实验目前正在进行中。

In the second year of this project, we have executed the Materials Transfer Agreements necessary for us to obtain the published chordoma cell lines, including a new cell lines JHC-1 and MUG-Chor1. In all four lines, using western blotting and immunoprecipitation, we have confirmed high-level expression of human T. Experiments have shown that chordoma cell line proliferation is significantly slowed by anti-T shRNA treatment. Appropriate assays confirm the downregulation of T mRNA and protein. cDNA microarray analysis is to compare cell lines treated with anti-T shRNA with control shRNA treated cell lines are currently underway and is expected to be completed within a month. Furthermore, the baseline microRNA environment of chordoma cells has been analyzed. That for cell lines downregulated for T by shRNA will soon be completed as well.Finally, we have also identified microRNA's that are altered in their expression after ionizing irradiation treatment of chordoma cell lines. cDNA microarray analysis of similar experiments shows a number of known, targetable genes that are modulated by ionizing radiation. Confirmatory experiments are underway at this time.