title: The miR-17-92 cluster regulates FOG-2 expression and inhibits proliferation of mouse embryonic cardiomyocytes.
abstract: MicroRNAs (miRNAs) have gradually been recognized as regulators of embryonic development; however, relatively few miRNAs have been identified that regulate cardiac development. A series of recent papers have established an essential role for the miRNA-17-92 (miR-17-92) cluster of miRNAs in the development of the heart. Previous research has shown that the Friend of Gata-2 (FOG-2) is critical for cardiac development. To investigate the possibility that the miR-17-92 cluster regulates FOG-2 expression and inhibits proliferation in mouse embryonic cardiomyocytes we initially used bioinformatics to analyze 3' untranslated regions (3'UTR) of FOG-2 to predict the potential of miR-17-92 to target it. We used luciferase assays to demonstrate that miR-17-5p and miR-20a of miR-17-92 interact with the predicted target sites in the 3'UTR of FOG-2. Furthermore, RT-PCR and Western blot were used to demonstrate the post-transcriptional regulation of FOG-2 by miR-17-92 in embryonic cardiomyocytes from E12.5-day pregnant C57BL/6J mice. Finally, EdU cell assays together with the FOG-2 rescue strategy were employed to evaluate the effect of proliferation on embryonic cardiomyocytes. We first found that the miR-17-5p and miR-20a of miR-17-92 directly target the 3'UTR of FOG-2 and post-transcriptionally repress the expression of FOG-2. Moreover, our findings demonstrated that over-expression of miR-17-92 may inhibit cell proliferation via post-transcriptional repression of FOG-2 in embryonic cardiomyocytes. These results indicate that the miR-17-92 cluster regulates the expression of FOG-2 protein and suggest that the miR-17-92 cluster might play an important role in heart development.
authors.affiliation: Department of Cardiology, First Affiliated Hospital, Chongqing Medical University, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Xiang
authors.forename: Han
authors.initials: H
authors.lastname: Lei
authors.forename: Mianzhi
authors.initials: M
authors.lastname: Chen
authors.forename: Qinwei
authors.initials: Q
authors.lastname: Li
authors.forename: Huan
authors.initials: H
authors.lastname: Sun
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Ai
authors.forename: Tielin
authors.initials: T
authors.lastname: Chen
authors.forename: Honglian
authors.initials: H
authors.lastname: Wang
authors.forename: Yin
authors.initials: Y
authors.lastname: Fang
authors.forename: Qin
authors.initials: Q
authors.lastname: Zhou
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics*; D000818:Animals; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D019295:Computational Biology; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D005260:Female; D008156:Luciferases / Q000494:pharmacology; D051379:Mice; D008822:Mice, Transgenic; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology*; D010957:Plasmids / Q000235:genetics; D011247:Pregnancy; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
chemical_list: D020413:3' Untranslated Regions; D004268:DNA-Binding Proteins; D035683:MicroRNAs; D014157:Transcription Factors; C492170:Zfpm2 protein, mouse; D008156:Luciferases