Core Promoter Analysis Of Porcine Six1 Gene And Its Regulation Of The Promoter Activity By Cpg Methylation

Six1, an evolutionary conserved transcription factor, has been shown to play an important role in organogenesis and diseases. However, no reports were shown to investigate its transcriptional regulatory mechanisms. In the present study, we first identified porcine Sixl gene core promoter region (+ 170/-360) using luciferase reporter assay system and found that promoter activities were significantly higher in the mouse myoblast C2C12 cells than that in the mouse fibroblast C3H10T1/2 cells, implying that Six] promoter could possess muscle-specific characteristics. Moreover, our results showed that promoter activities of Sixl were decreased as induction of differentiation of C2C12 cells, which was accompanied by the down-regulation of mRNA expression of Six) gene. In addition, we found that the DNA methylation of Six) promoters in vitro obviously influences the promoter activities and the DNA methylation level of Six) promoter core region was negatively correlated to Six) gene expression in vivo. Taken together, we preliminarily clarified transcriptional regulatory mechanisms of Six) gene, which should be useful for investigating its subtle transcriptional regulatory mechanisms in the future. On the other hand, based on Six) involved in tumorigenesis, our data also provide a genetic foundation to control the generation of diseases via pursuing Six1 as therapeutic target gene. (C) 2013 Elsevier B.V. All rights reserved.