Affiliation:
1. College of Animal Science and Technology, Shihezi University, Shihezi 832000, China
Abstract
Background:
microRNAs (miRNAs) are a small, endogenous non-coding RNAs that are
involved in post-transcriptional gene regulation of many biological processes, including embryo
implantation and placental development. In our previous study, miR-146a-5p was found expressed
higher in the serum exosomes of pregnant sows than non-pregnant. The research on miR-146a-5p
has been mainly related to human diseases, but there are few studies on its effects on the
reproduction of sows in early pregnancy.
Objective:
In this article, our motivation is to study the role of miR-146a-5p in the early pregnancy
of sows on the cell proliferetion and apoptosis by targeting SMAD3 and SMAD4.
Methods:
Bioinformatics software was used to identify the target genes of miR-146a-5p. The wildtype
and mutant-type recombinant plasmids of dual-luciferase reporter with 3'-UTR of Smad3 or 3'-
UTR of Smad4 were constructed, and co-transfected in porcine kidney cell (PK-15 cell) with miR-
146a-5p mimic, mimic-NC(M-NC), inhibitor and inhibitor-NC(IN-NC), then dual-luciferase
activity analysis, qRT-PCR and Western blot were performed to verify the target genes. After the
transfection of BeWo choriocarcinoma cell (BeWo cell) with miR-146a-5p mimic, M-NC, inhibitor
and IN-NC, the mRNA expression of Caspase-3, BAX and Bcl-2 was measured using qRT-PCR,
and the cell proliferation was measured using CCK-8 kit.
Results:
The luciferase, mRNA and protein expression of Smad3 in PK-15 cells treated by Smad3-
3'-UTR-W co-transfected with miR-146a-5p mimic were significantly lower than that with miR-
146a-5p M-NC, and the results of Smad4 were similar to Smad3, but the protein expression had a
trend to lower in mimic group. The expression level of Bcl-2 in the miR-146a-5p mimic group was
significantly lower than that in the miR-146a-5p M-NC group, but the expression pattern of
Caspase-3 was just opposite. The mimic of miR-146a-5p reduced the proliferation of BeWo cells,
however the inhibitor increased.
Conclusion:
Smad3 and Smad4 are the direct target genes of miR-146a-5p. The expression of
Smad3 and Smad4 were affected by the mimic and inhibitor of miR-146a-5p. miR-146a-5p affects
cell apoptosis and proliferation by regulating their target genes. This study provided new data to
understand the regulation mechanism of early pregnancy in sows.
Funder
National Natural Science Foundation of China
Publisher
Bentham Science Publishers Ltd.
Subject
Biochemistry,General Medicine,Structural Biology
Cited by
11 articles.
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