Identification of the potential target genes of microRNA-146a induced by PMA treatment in human microvascular endothelial cells.
Phorbol 12-myristate 13-acetate (PMA) is known to activate protein kinase C (PKC) and increase angiogenesis in cultured endothelial cells. Using a microRNA (miRNA) array, we found that PMA induced miR-146a expression in human microvascular endothelial cells. The miR-146a expression was dependent on dose and time and independent of PKC activation. Using a combined approach involving predictions using miRanda algorithm and whole genome microarray experiments with or without inhibition of miR-146a expression by LNA-antimir-146a or LNA-control, 29 potential target genes of miR-146a were identified. Because endothelial cell S phase progression is an early event in the induction of angiogenesis, we evaluated 5 cell cycle-related genes from the 29 target genes and found that the transcripts of 3 genes (CCNA2, PA2G4, and BRCA1) were downregulated after PMA treatment, but their expression was rescued upon miR-146a inhibition. However, inhibition of miR-146a expression failed to alter the cell cycle distribution or angiogenesis induced by PMA treatment. By using a combined approach involving computational prediction and a whole genome microarray experiment in the presence or absence of antimir, the observations in this presented article raise the possibility that antimir strategies might be used to identify the potential miRNA targets.
Hsieh
Ching-Hua
CH
Department of Plastic and Reconstructive Surgery, Chang Gung Memorial Hospital, Kaohsiung Medical Center, Chang Gung University College of Medicine, Kaohsiung Hsien, Taiwan. m93chinghua@livemail.tw
Rau
Cheng-Shyuan
CS
Jeng
Seng-Feng
SF
Lin
Chia-Jung
CJ
Chen
Yi-Chun
YC
Wu
Chia-Jung
CJ
Lu
Tsu-Hsiang
TH
Lu
Cheng-Hsien
CH
Chang
Wen-Neng
WN
D002453:Cell Cycle / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D055785:Gene Knockdown Techniques; D006801:Humans; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D055806:Microvessels / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism; D018919:Neovascularization, Physiologic / Q000187:drug effects / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D013997:Time Factors
C542542:MIRN146 microRNA, human; D035683:MicroRNAs; D013755:Tetradecanoylphorbol Acetate