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Screening Small Anti-leukemia Compounds Based on EDAG
Author: DiZuo
Tutor: WangSiYing
School: Anhui Medical University,
Course: Pathology and Pathophysiology
Keywords: EDAG Transcriptional activation The dual luciferase Report system Drug screening
CLC: R733.7
Type: Master's thesis
Year: 2007
Downloads: 19
Quote: 0
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Abstract
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Objective: To establish based EDAG ( erythroid developmental gene erythoid develop associated gene ) transcriptional activity of new small molecule compounds in vitro screening model and use the model to screen small molecule compound libraries , expect to quickly and efficiently find the pilot molecules with potential anti-leukemia . Methods: The experimental method of the luciferase reporter gene to The EDAG the transcriptional activation quantify establish the model of large-scale screening of active small molecule compounds and screening of small molecule compound libraries (about 1000 ) . Tetrazolium MTS colorimetric , 3H-TdR incorporation assay , flow cytometry screening active compounds to high expression of endogenous the EDAG the leukemia cell proliferation , apoptosis and differentiation ; application of signal transduction pathways specific inhibitor (U0126, CalphostinC, LY294002) the influence of the test compound on the EDAG luciferase reporter gene system , and further verified by Western blot , and thus the mechanism of the candidate active compound obtained in preliminary discussion . Results: pM-EDAG expression vector was successfully constructed , and the establishment of a high-throughput screening model , the application of the model obtained by screening of candidate active compounds 2G2 and 2F10 . 2G2 inhibition of EDAG transcriptional activation of specific inhibition of K562 cell proliferation , promote apoptosis , but no significant effect on their differentiation ; 2F10 enhance transcriptional activation activity of EDAG promote megakaryocytic differentiation of K562 cells were , did not significantly inhibit proliferation and promote apoptosis , MAPK signal transduction pathway inhibitor U0126 2F10 transcriptional activation effect specific inhibition . Conclusion: This study successfully established a high - throughput screening of the EDAG transcription activity influential model of the active compounds , the preliminary study of the function and mechanism of action of the active compounds . Lead compounds for further reconstructive , may provide a new target for the treatment of leukemia .
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CLC: > Medicine, health > Oncology > Hematopoietic and lymphoid neoplasms > Leukemia
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