Analysis of BRD4-regulated genes in human glioma U251 cells. Homo sapiens

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 Overall design: 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.