人神经母细胞瘤株中β-淀粉样前体蛋白基因IL-1反应元件的研究

细胞团子IL-1β可以诱导SH-SY5Y神经母细胞瘤细胞中APP基因的转录．为了研究APP基因启动子区的IL-1β反应元件,用含不同长度APP启动子片段的报告基因载体,瞬时转染SH-SY5Y细胞,发现APP启动子在SH-SY5Y细胞中有较强活性,其中-488到-303区为IL-1β诱导APP转录活性增加所必需,这个区域包括1个AP1结合位点和1个HSE元件．     

Identification of the RA response element and transcriptional silencer in human <Emphasis Type="Italic">αCaMKII</Emphasis> promoter

The promoter of alpha subunit of the rat calcium/calmodulin-dependent protein kinase II (alphaCaMKII) gene was identified to contain an essential TATA element. Cell-based functional assay showed that the rat promoter displayed greater activity in neuronal cells than in non-neuronal cells. To characterize the human alphaCaMKII promoter, we have developed a promoter-reporter gene assay using different cell lines. A 2047 base pairs (bp) human alphaCaMKII gene promoter was cloned from human genomic DNA. Unlike the rat alphaCaMKII promoter, DNA sequence analysis showed that the human promoter was devoid of TATA element. We made series deletions of the promoter and fused the different sizes of the human promoter sequences to a luciferase reporter gene. The promoter-reporter constructs were transfected into human neuroblastoma SH-SY5Y, human neuroblastoma BE(2)-M17, and rat pheochromocytoma PC12 neuronal cell lines as well as human embryonic kidney HEK293 and human glioma U251 non-neuronal cell lines. The reporter gene assay demonstrated that the human alphaCaMKII promoter displayed high activity in the neuronal cell lines, while the activity was low in non-neuronal cell lines. All-trans retinoic acid (RA) enhanced the promoter activity in SH-SY5Y cells. Further analysis showed that there were two RA response elements located between +11 and +136 and -1911 to -593. In addition, we have identified a potent silencer at position -179 to -244 of the human alphaCaMKII promoter.