title: miR181c promotes apoptosis and suppresses proliferation of metanephric mesenchyme cells by targeting Six2 in vitro.
abstract: Increasingly recognized importance has been assumed for microRNA (miRNA) in the regulation of the delicate balance of gene expression. In our study, we aimed to explore the regulation role of miR181c towards Six2 in metanephric mesenchyme (MM) cells. Bioinformatics analysis, luciferase assay and semi-quantitative real-time (RT) PCR, subsequently RT PCR, Western blotting, 5-ethynyl-2'-deoxyuridine cell proliferation assay, Cell Counting Kit-8 assay, immunofluorescence and flow cytometry, were employed to verify the modulation function of miR181c on Six2 in the mK3 MM cell line that is one kind of MM cells. miR181c was predicted to bind the 3' untranslated region of Six2 by bioinformatics analysis, which was subsequently validated by the in vitro luciferase reporter assay. Moreover, transfection of miR181c mimic can decrease the expression of Six2 both in mRNA and protein levels in mK3 cells. Still, ectopic expression of miR181c inhibits the proliferation, promotes the apoptosis and even makes the nephron progenitor phenotype lose mK3 cells. These results revealed the ability of a single miRNA-miR181c to downregulate the expression of Six2, restrain the proliferation and promote the apoptosis that even makes the nephron progenitor phenotype lose MM cells, suggesting a potential role of miR181c during the kidney development.
authors.affiliation: Department of Dermatology, West China Hospital, West China School of Medicine, Sichuan University, Chengdu, China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Lv
authors.forename: Zhaomin
authors.initials: Z
authors.lastname: Mao
authors.forename: Zhongshi
authors.initials: Z
authors.lastname: Lyu
authors.forename: Pengzong
authors.initials: P
authors.lastname: Zhang
authors.forename: Alan
authors.initials: A
authors.lastname: Zhan
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
authors.forename: Hui
authors.initials: H
authors.lastname: Yang
authors.forename: Mi
authors.initials: M
authors.lastname: Li
authors.forename: Honglian
authors.initials: H
authors.lastname: Wang
authors.forename: QianYa
authors.initials: Q
authors.lastname: Wan
authors.forename: Hongyuan
authors.initials: H
authors.lastname: Wei
authors.forename: Ming
authors.initials: M
authors.lastname: Wang
authors.forename: Nian
authors.initials: N
authors.lastname: Wang
authors.forename: Xun
authors.initials: X
authors.lastname: Li
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
authors.forename: Hui
authors.initials: H
authors.lastname: Zhao
authors.forename: Qin
authors.initials: Q
authors.lastname: Zhou
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D049109:Cell Proliferation; D019295:Computational Biology; D015536:Down-Regulation; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D007668:Kidney / Q000166:cytology* / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D009399:Nephrons / Q000166:cytology / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D020413:3' Untranslated Regions; D018398:Homeodomain Proteins; D035683:MicroRNAs; D012333:RNA, Messenger; C497420:Six2 protein, mouse; D014157:Transcription Factors; C529100:mirn181 microRNA, mouse
keywords: Six2; apoptosis; kidney; microRNA; proliferation