Study On Regulation Of Human Notch4 Gene Transcription

AIM To investigate the effects of drugs and cytokines on Notch4 signaling and regulation. It will open a new way of molecular pharmacology for regulating the Notch4 gene on cell proliferation and differentiation.METHODS The Notch4 gene expression was studied by real time RT-PCR, and the effects of drugs and cytokines were screened on Nocth4 gene expression in HUVEC and HL-60 cells. Using analytical cytology and molecular biology, the MTT assay,flow cytometry, real time-RT PCR and western blot, The effects of alltans retinoic acid (ATRA) and arsenic trioxide (As) were investigated on cell proliferation, differentiation and the expression of Notch4 and Notch1 signaling pathways in HL-60 cells. The Notch4 promoter of human cord blood CD34 + hematopoietic stme/progenitor cell was amplified by polymerase chain reaction, and GFP report vectors of Notch4 promoter were reconstructed by genetic recombination technology. Expression methods of recombinant vectors were observed by flow cytometry and fluorescent microscope in fluorescence and bright field in cells transfected with GFP-Notch 4 plasmid.