Friedrich Inbar, Eizenbach Menahem, Sajman Julia, Ben-Bassat Hannah, Levitzki Alexander
Unit of Cellular Signaling, Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Givat Ram, Jerusalem 91904, Israel.
Mol Ther. 2005 Nov;12(5):969-75. doi: 10.1016/j.ymthe.2005.06.442. Epub 2005 Aug 8.
Long double-stranded RNA (>30 bp), usually expressed in cells infected with RNA viruses, triggers antiviral responses that induce apoptosis of the infected cells. PKR can be selectively activated in glioblastoma cells by in situ generation of dsRNA following introduction of antisense RNA complementary to an RNA expressed specifically in these cells. Harnessing PKR for the selective killing of cancer cells is potentially a powerful strategy for treating cancer, but we were unable to induce apoptosis by this approach in a T cell lymphoma. We therefore established a cellular screening assay to test the ability of PKR to induce death in cell lines, especially those originating from human cancers. This "PKR killing screen" is based on the infection of cells with an adenoviral vector encoding GyrB-PKR, followed by coumermycin treatment. Cancers represented by cell lines in which PKR activation leads to cell death are good candidates for the dsRNA killing approach, using antisense to RNA molecules specifically expressed in these cells. The PKR killing screen may also serve as a tool for exploring PKR signaling and other related pathways, by identifying new cases in which PKR signaling is inhibited or impaired.
长双链RNA（>30bp）通常在感染RNA病毒的细胞中表达，可触发抗病毒反应，诱导被感染细胞凋亡。通过导入与这些细胞中特异性表达的RNA互补的反义RNA，可在原位产生双链RNA，从而在胶质母细胞瘤细胞中选择性激活PKR。利用PKR选择性杀伤癌细胞可能是一种强大的癌症治疗策略，但我们无法通过这种方法在T细胞淋巴瘤中诱导凋亡。因此，我们建立了一种细胞筛选试验，以测试PKR在细胞系（尤其是源自人类癌症的细胞系）中诱导死亡的能力。这种“PKR杀伤筛选”基于用编码GyrB-PKR的腺病毒载体感染细胞，随后进行香豆霉素处理。PKR激活导致细胞死亡的细胞系所代表的癌症，是使用针对这些细胞中特异性表达的RNA分子的反义RNA进行双链RNA杀伤方法的良好候选对象。通过识别PKR信号传导被抑制或受损的新情况，PKR杀伤筛选也可作为探索PKR信号传导和其他相关途径的工具。
