关键词: 神经前体细胞, 人端粒酶反转录酶, 启动子, 转录, 荧光素酶检测

Abstract: Objective Using hTERT-immortalized human neural progenitor cell line, hNPCs-G3, it was studied on the transcription activity of endogenous human telomerase reverse transcriptase(hTERT) gene and the mechanisms of proliferation and differentiation in the human neural progenitor cells. Methods Transient co-transfection of hTERT promoter-luciferase constructs and pSV-β-Gal control vector into HeLa cells and hNPCs-G3 cells, and detection of the transcription activities of different hTERT promoter-luciferase constructs. After co-tansfection as above, hNPCs-G3 cells were induced by bFGF, and the transcription activities of different length of hTERT promoters responsive to proliferation were analyzed. After the co-tansfection as above, hNPCs-G3 cells were induced by cAMP, and the transcription activities of different length of hTERT promoters responsive to differentiation were analyzed. Results The transcription activity of endogenous hTERT gene was low in human neural progenitor cells, and activity of the proximal promoter was the highest. bFGF could up-regulate the activitiy of endogenous hTERT gene in human neural progenitor cells, which mainly depended on the stretch of -2 098bp to -1 099bp and -3 216bp to -2 098bp upstream of ATG. cAMP could downregulate the transcription activity of endogenous hTERT gene in human neural progenitor cells, which inhibited the activity of proximal promoter.Conclusion The transcription activity of endogenous hTERT gene was low in human neural progeni

Key words: Neural progenitor cell, Human telomerase reverse transcriptase, Promoter, Transcription, Luciferase assay