Construction of a drug screening model based on transcription regulation of STAT3 / 中国药理学通报

ABSTRACT

AIM: To find small molecule organic mimetics of G-CSF, according to the transcription regulating effect of the important transcription factors-STAT3 (signal transducers and activators of transcription3) in hematopoiesis growth factor's signal transductional passageway, we established a drug screening model targeting transcription regulating of STAT3. METHODS: A recombinat vector pTKS3-CAT was constructed. Then pTKS3-CAT and pTK-Hyg were transfected into NFS-60 cells expressing G-CSF receptor with lipofectamine 2000. Cells derived from hygromycin-resistant colonies were tested for CAT activity, and positive colonies were isolated. At the same time, the sensitivity and the specialty of the model were tested. RESULTS: A dose-dependent expression of CAT gene with half-maximal induction by rhG-CSF at 0.044 nmol·L-1 was observed. After treating with rhEPO CAT activity couldn't be tested. CONCLUSION: The expression of CAT gene could be strongly induced by its special ligands in drug screening model. This model can be used to assay CAT from extracts of cells grown with CAT-ELISA method in microtiter wells and then to screen small molecular compounds with G-CSF-like activity.