Characterization Of HMGA1 Promoter Activity Regions

Objective: The aim of this study was to identify the core activity region of HMGA1 promoter in HUVEC-12 and MCF-7 cells, and to uncover the effect of HMGA1 on high glucose induced apoptosis in HUVEC-12 cells.Motheds: We generated various 5'-3'deletion luciferase reporter constructs of HMGA1 DNA fragment relative to a putative transcriptional start site, with the method of PCR amplification. Dual-luciferase activity assay was performed to assess the activity of HMGA1 promoter in HUVEC-12 and MCF-7 cells respectively. The ORF region of HMGA1 gene was amplified and ligated with pCDNA3.1(-) plasmid, and pCDNA3.1(+)/HMGA1 was trasnfected into HUVEC-12 cells.Results: Functional analysis of the constructs showed that a proximal promoter region (-256/+1) of HMGA1 maintained high level of basal activity in MCF-7 cells and the proximal promoter region (- 598 /+1) showed a high level of basal activity in HUVEC-12 cells. High glucose induced HUVEC-12 cells apoptosis could be inhibited by the ectopic HMGA1 expression.Conclusions: The proximal promoter region (- 598 /+1) of HMGA1 could be the core sequence for the activity of HMGA1 promoter and the cellular context contribute to the HMGA1 promoter activity. HMGA1 expression blocks the high glucose induced HUVEC-12 cells apoptosis.