Potential transcriptional regulatory regions exist upstream of the human ezrin gene promoter in esophageal carcinoma cells

We previously demonstrated that the region −87/+134 of the human ezrin gene (VIL2) exhibited promoter activity in human esophageal carcinoma EC109 cells, and a further upstream region −1324/−890 positively regulated transcription. In this study, to identify the transcriptional regulatory regions upstream of the VIL2 promoter, we cloned VIL2 −1541/−706 segment containing the −1324/−890, and investigated its transcriptional regulatory properties via luciferase assays in transiently transfected cells. In EC109 cells, it was found that VIL2 −1541/−706 possessed promoter and enhancer activities. We also localized transcriptional regulatory regions by fusing 5′- or 3′-deletion segments of VIL2 −1541/−706 to a luciferase reporter. We found that there were three positive and one negative transcriptional regulatory regions within VIL2 −1541/−706 in EC109 cells. When these regions were separately located upstream of the luciferase gene without promoter, or located upstream of the VIL2 promoter or SV40 promoter directing the luciferase gene, only VIL2 −1297/−1186 exhibited considerable promoter and enhancer activities, which were lower than those of −1541/−706. In addition, transient expression of Sp1 increased ezrin expression and the transcriptional activation of VIL2 −1297/−1186. Other three regions, although exhibiting significantly positive or negative transcriptional regulation in deletion experiments, showed a weaker or absent regulation. These data suggested that more than one region upstream of the VIL2 promoter participated in VIL2 transcription, and the VIL2 −1297/−1186, probably as a key transcriptional regulatory region, regulated VIL2 transcription in company with other potential regulatory regions.