title: Small molecule inhibitor of myogenic microRNAs leads to a discovery of miR-221/222-myoD-myomiRs regulatory pathway.
abstract: Myogenic microRNAs (myomiRs) that are specifically expressed in cardiac and skeletal muscle are highly relevant to myogenic development and diseases. Discovery and elucidation of unknown myomiRs-involved regulatory pathways in muscle cells are important, but challenging due to the lack of proper molecular tools. We report here a miR-221/222-myoD-myomiRs regulatory pathway revealed by using a small-molecule probe that selectively inhibits myomiRs including miR-1, miR-133a, and miR-206. The small-molecule inhibitor screened from luciferase assay systems was found to inhibit myomiRs and differentiation of C2C12 cells. Using the small molecule as a probe, we found that the transcriptional factor myoD, which is upstream of myomiRs, was further regulated by miR-221/222. This miR-221/222-myoD-myomiRs regulatory pathway was confirmed by over-expressing or knockdown miR-221/222 in muscle cells, which respectively led to the inhibition or enhancement of myoD protein expression and subsequent downregulation or upregulation of myomiR expression.
authors.affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, PRC; School of Life Science, State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, PRC.
authors.forename: Su-Bee
authors.initials: SB
authors.lastname: Tan
authors.affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, PRC.
authors.forename: Jinbo
authors.initials: J
authors.lastname: Li
authors.affiliation: School of Life Science, State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, PRC.
authors.forename: Xi
authors.initials: X
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, PRC.
authors.forename: Wenjie
authors.initials: W
authors.lastname: Zhang
authors.affiliation: School of Life Science, State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, PRC.
authors.forename: Dianmu
authors.initials: D
authors.lastname: Zhang
authors.affiliation: School of Life Science, State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, PRC. Electronic address: cyzhang@nju.edu.cn.
authors.forename: Chenyu
authors.initials: C
authors.lastname: Zhang
authors.affiliation: School of Life Science, State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, PRC. Electronic address: donghaili@nju.edu.cn.
authors.forename: Donghai
authors.initials: D
authors.lastname: Li
authors.affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, PRC. Electronic address: njuzy@nju.edu.cn.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D020413:3' Untranslated Regions; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D015870:Gene Expression / Q000187:drug effects; D057809:HEK293 Cells; D006801:Humans; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism; D017570:MyoD Protein / Q000737:chemistry / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism; D020320:Oligoribonucleotides, Antisense / Q000378:metabolism; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D054852:Small Molecule Libraries / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D014162:Transfection
chemical_list: D020413:3' Untranslated Regions; D035683:MicroRNAs; D017570:MyoD Protein; D020320:Oligoribonucleotides, Antisense; D012333:RNA, Messenger; D054852:Small Molecule Libraries; C525709:MIR221, human; C525710:MIR222, human