Screening Small Anti-leukemia Compounds Based On EDAG

Objective: To set up an in vitro high-throughput drug screening model targeting transcription regulating of Erythroid differentiation-associated gene (EDAG) and to screen a small molecule library by this model, We hope to find small molecule compounds quickly and efficiently, according to the transcription regulating effect of EDAG .Methods: As EDAG has the activity of transcriptional activation, we set up a new model for large scaled screening small molecular compounds on 96-well plate by luciferase reporter assay.In this model, and screen small molecular compound library (1000). we do some initial research on the function these 2 screened compounds: the cellular proliferation was detected by MTS and growth curve assay , the distributions of cell cycles were detected by flow cytometry , using the specific inhibitor(U0126,CalphostinC,LY294002) of the signal transduction by the luciferase reporter gene assay system, attempts to find the function mechanism of the compounds and further confirms with Western blot.Results: The eukaryotic expression vector, pM-EDAG, is constructed successfully.We set up an in vitro high-throughput drug screening model. Through this method, we get 2 potential compounds, 2G2,2F10, which affect EDAG transcriptional regulation. 2G2 small compound could repress transcriptional activity of EDAG, whereas 2F10 could activate transcriptional activity of EDAG. Furthermore, 2G2 could inhibit the proliferation of K562 cells which high express EDAG. and the cell cycle G2/M hinder. 2F10 leads to K562 megacaryocyte differentiation is the split up function, not the obvious inhibit the proliferation of K562 cells. The function mechanism possibly is leads the MAPK signal transduction pathway.Conclusion: We established the drug screening model targeting transcription regulating of EDAG successfully and use this model to screen a small molecule library. 2G2 which repress transcriptional activity of EDAG, then through structure transformation, maybe relate to the diagnosis and treatment of leukemia.