Analysis of BRD4-regulated genes in human glioma U251 cells

BRD4 is a member of the bromodomain and extra-terminal (BET) family, which has become a promising drug target for numerous cancers. It has been reported that BRD4 is deregulated in gliomas; however, the precise molecular pathways regulated by BRD4 are still elusive. In this study, we knocked down BRD4 expression in glioma cell line U251 by lentivirus-mediated short hairpin RNA (shRNA) approach. BRD4 downregulation attenuated cell proliferation and promoted cell apoptosis. Genome-wide analysis of BRD4-regulated transcripts in U251 cells was performed using microarray to reveal the possible molecular mechanism To knock down BRD4 expression in U251 cells, lentivirus that expressed shRNA against human BRD4 were transfected U251 cells. Scrambled shRNA (Scr-shRNA) that targeted a non-specific sequence was used as the normal control. The efficiency of BRD4 knockdown were demonstrated by real-time PCR and western blot assays. BRD4-shRNA- transfected and Scr-shRNA-transfected U251 cells were collected for RNA extraction and hybridization on Affymetrix microarrays.