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Involvement of miR‐140‐3p in Wnt3a and TGF β3 signaling pathways during osteoblast differentiation in MC 3T3‐E1 cells
Author(s) -
Fushimi Shigeko,
Nohno Tsutomu,
Nagatsuka Hitoshi,
Katsuyama Hironobu
Publication year - 2018
Publication title -
genes to cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.912
H-Index - 115
eISSN - 1365-2443
pISSN - 1356-9597
DOI - 10.1111/gtc.12591
Subject(s) - runx2 , wnt3a , osteoblast , wnt signaling pathway , biology , signal transduction , transcription factor , transfection , microbiology and biotechnology , transforming growth factor , microrna , gene expression , cellular differentiation , gene , genetics , in vitro
The Wnt/β‐catenin signaling and TGF β signaling pathways play a key role in osteoblast differentiation. The mi RNA s play important roles in regulating gene expression at the post‐transcriptional level through fine‐tuning of protein‐encoding gene expression. However, involvement of mi RNA s is not established for Wnt3a and TGF β signaling pathways in osteoblast differentiation. Here, we examined the role of mi RNA s expressed differentially after Wnt3 a expression during osteoblast differentiation. Over‐expression of the Wnt3a gene increased ALP transcription, but decreased Col1 , Runx2, and OCN transcription in osteoblastic MC 3T3‐E1 cells. Expression profiling and quantitative PCR for mi RNA s showed that miR‐140‐3p decreased in Wnt3a ‐over‐expressing osteoblastic cells. Wnt3a over‐expression increased TGF β3 expression, whereas transfection of the miR‐140‐3p mimic into MC 3T3‐E1 cells significantly inhibited TGF β3 expression. Luciferase assay for the TGF β 3 transcript showed that TGF β 3 was a direct target of miR‐140‐3p. miR‐140‐3p mimic transfection resulted in significantly increased OCN transcription, but did not affect ALP , Col1 , and Runx2 transcription in MC 3T3‐E1 cells. rTGF β3 treatment decreased OCN transcription in MC 3T3‐E1 cells. These results suggest that the miR‐140‐3p is involved in osteoblast differentiation as a critical regulatory factor between Wnt3a and TGF β3 signaling pathways.

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