Crosstalk between DNA methylation and histone acetylation triggers GDNF high transcription in glioblastoma cells

摘要

High CREB expression significantly promotes transcription in U251 GBM cells. Putative CRE sites within the human promoter II. Arrows indicate the major TSS corresponding to the promoter II. The TSS was considered + 1. The letter E indicates enhancer II, while the letter S indicates silencer II. Comparison of mRNA levels (log median-centered ratio) of human CREB in NB samples ( = 10) and glioblastoma brain samples ( = 542) from TCGA brain dataset analyzed on the Oncomine® Platform. mRNA. Protein expression in NB tissue and low- and high-grade GBM tissues as determined by real-time PCR and western blot ( = 3). mRNA. Protein expression in NHA cells and GBM cell lines (U251 and U343) ( = 3). mRNA. Protein expression after 72 h infection with CREB-KDs lentivirus in U251 cells ( = 3). mRNA. Protein expression after 72 h infection with CREB-KD1 in U251 cells ( = 3). U251 cells were transfected with the pGDNF-Luc(− 1300/+ 149)-CRE-WT and pRL-TK plasmids after 24 h of CREB-KD1 infection; and the effect of CREB expression on the promoter II activity was assessed by dual-luciferase assay after another 48 h of incubation ( = 3). mRNA expression after 72 h infection with CREB-OE and KCREB-OE in U251 cells ( = 3). Binding of CREB to CRE in enhaner II (CRE-E) of promoter II after 72 h infection with CREB-OE lentivirus in U251 cells as determined by ChIP-PCR ( = 3). was used as an internal control. All data except in ( are mean ± SD. ** < 0.01