Proliferation of metanephric mesenchymal cells is inhibited by miR-743a-mediated WT1 suppression in vitro.
To seek out the potential microRNAs (miRNAs) that target Wilms' tumor suppressor 1 (WT1), a transcription factor required for progenitor proliferation as well as normal development of the kidney, and to clarify the effects of the miRNAs on WT1, the 3'-untranslated region (3'‑UTR) of WT1 was initially analyzed and miR‑743a, a seldom‑reported miRNA, was identified. In the present paper, luciferase reporter assays were performed to confirm that miR‑743a is able to directly target the 3'‑UTR of WT1. Subsequently, reverse transcription‑quantitative polymerase chain reaction, combined with western blotting analyses, were performed, and the results revealed a significant inhibition of WT1 at the mRNA and the protein levels. Furthermore, a 5‑ethynyl‑2'‑deoxyuridine (EdU) cell proliferation assay, coupled with a WT1 rescue strategy, demonstrated that miR‑743a inhibited the proliferation of metanephric mesenchymal (MM) cells, in part by targeting WT1. In conclusion, by targeting WT1, miR‑743a suppresses the proliferation of MM cells in vitro, and probably possesses vital functions in kidney development and kidney‑associated diseases.
Division of Molecular Nephrology and The Creative Training Center for Undergraduates, M.O.E. Key Laboratory of Laboratory Medical Diagnostics, College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, P.R. China.
Mingming
M
Xue
The Seventh Class of 2012 Year Entry, Third Clinical College, Chongqing Medical University, Chongqing 400016, P.R. China.
Yuru
Y
Zhou
Division of Molecular Nephrology and The Creative Training Center for Undergraduates, M.O.E. Key Laboratory of Laboratory Medical Diagnostics, College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, P.R. China.
Xiaoyu
X
Liu
Division of Molecular Nephrology and The Creative Training Center for Undergraduates, M.O.E. Key Laboratory of Laboratory Medical Diagnostics, College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, P.R. China.
Dongsheng
D
Ni
Division of Molecular Nephrology and The Creative Training Center for Undergraduates, M.O.E. Key Laboratory of Laboratory Medical Diagnostics, College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, P.R. China.
Yanxia
Y
Hu
Division of Molecular Nephrology and The Creative Training Center for Undergraduates, M.O.E. Key Laboratory of Laboratory Medical Diagnostics, College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, P.R. China.
Yaoshui
Y
Long
Division of Molecular Nephrology and The Creative Training Center for Undergraduates, M.O.E. Key Laboratory of Laboratory Medical Diagnostics, College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, P.R. China.
Pan
P
Ju
Division of Molecular Nephrology and The Creative Training Center for Undergraduates, M.O.E. Key Laboratory of Laboratory Medical Diagnostics, College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, P.R. China.
Qin
Q
Zhou
D020413:3' Untranslated Regions; D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation / Q000235:genetics; D006801:Humans; D007668:Kidney / Q000254:growth & development / Q000378:metabolism* / Q000473:pathology; D007674:Kidney Diseases / Q000235:genetics / Q000473:pathology; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D051379:Mice; D035683:MicroRNAs / Q000096:biosynthesis / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis; D012097:Repressor Proteins / Q000096:biosynthesis* / Q000235:genetics; D025721:WT1 Proteins
D020413:3' Untranslated Regions; D035683:MicroRNAs; C000616936:Mirn743 microRNA, human; C560968:Mirn743 microRNA, mouse; D012333:RNA, Messenger; D012097:Repressor Proteins; D025721:WT1 Proteins; C000602896:WT1 protein, mouse