MiR-181b targets Six2 and inhibits the proliferation of metanephric mesenchymal cells in vitro.
MicroRNAs (miRNAs) are small non-coding RNAs that down-regulate gene expression by binding to target mRNA for cleavage or translational repression, and play important regulatory roles in renal development. Despite increasing genes have been predicted to be miRNA targets by bioinformatic analysis during kidney development, few of them have been verified by experiment. The objective of our study is to identify the miRNAs targeting Six2, a critical transcription factor that maintains the mesenchymal progenitor pool via self-renewal (proliferation) during renal development. We initially analyzed the 3'UTR of Six2 and found 37 binding sites targeted by 50 putative miRNAs in the 3'UTR of Six2. Among the 50 miRNAs, miR-181b is the miRNAs predicted by the three used websites. In our study, the results of luciferase reporter assay, realtime-PCR and Western blot demonstrated that miR-181b directly targeted on the 3'UTR of Six2 and down-regulate the expression of Six2 at mRNA and protein levels. Furthermore, EdU proliferation assay along with the Six2 rescue strategy showed that miR-181b suppresses the proliferation of metanephric mesenchymal by targeting Six2 in part. In our research, we concluded that by targeting the transcription factor gene Six2, miR-181b inhibits the proliferation of metanephric mesenchymal cells in vitro and might play an important role in the formation of nephrons.
The Division of Molecular Nephrology and the Creative Training Center for Undergraduates, The M.O.E. Key Laboratory of Laboratory Medical Diagnostics, The College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, PR China.
Zhongshi
Z
lyu
Zhaomin
Z
Mao
Honglian
H
Wang
Yin
Y
Fang
Tielin
T
Chen
Qianya
Q
Wan
Ming
M
Wang
Nian
N
Wang
Jiangming
J
Xiao
Hongyuan
H
Wei
Xun
X
Li
Yi
Y
Liu
Qin
Q
Zhou
D020413:3' Untranslated Regions; D000595:Amino Acid Sequence; D000818:Animals; D049109:Cell Proliferation; D057809:HEK293 Cells; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009399:Nephrons / Q000166:cytology / Q000254:growth & development*; D012333:RNA, Messenger / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
D020413:3' Untranslated Regions; D018398:Homeodomain Proteins; D035683:MicroRNAs; D012333:RNA, Messenger; C497420:Six2 protein, mouse; D014157:Transcription Factors; C529100:mirn181 microRNA, mouse
MicroRNAs; Proliferation; Six2; miR-181b